Hepatitis C virus infection in chronic haemodialysis patients, a clinicopathologic study. Fifty-two patients on regular haemodialysis at our institution were evaluated for the presence of HCV infection. Evaluation included detailed history, clinical examination, and monthly screening for anti-HCV antibody, liver enzymes (ALT, AST), serum iron and ferritin. Also, three-monthly screening for other viral markers, HBV (HBsAg, HBsAb, HBcAb), CMV (IgG and IgM), EBV, and HIV. Anti-HCV antibody was found in 21 patients (40.4%). There was a significant (P less than 0.05) relationship between presence of anti-HCV antibody and proportion of patients who received blood transfusion. During a 12-month follow-up, four (11.4%) patients seroconverted to be Anti-HCV positive while one case (4.8%) seroconverted to be anti-HCV negative. The frequency of elevation of liver enzymes was significantly higher in Anti-HCV positive cases (14/18) than in negative cases (11/28, P = 0.01). Evaluation of liver biopsies of 13 patients showed chronic persistent hepatitis in six and chronic active hepatitis in seven cases. We concluded that hepatitis C is a common problem among chronic haemodialysis patients at our institution; HCV infection is documented in 70% of all clinically diagnosed NANB hepatitis. Presence of anti-HCV antibodies cannot differentiate between active and past infection and cases with early HCV infection can be missed when relying on the mere detection of anti-HCV antibodies.


Long-term results of trabeculectomy with collagen sponge implant containing low-dose antimetabolite. Eighteen eyes of 15 patients with uncontrolled glaucoma (neovascular, intracapsular aphakia, uveitic, previous filtration failure) at high risk for failure of standard filtration underwent trabeculectomy with implantation of a purified collagen sponge containing 100 micrograms of antimetabolite (5-fluorouracil or bleomycin). Follow-up ranged from 3 months to 5 years. At the end of follow-up, 14 eyes (78%) had intraocular pressure below 21 mmHg and functional filtration blebs. No corneal toxicity was encountered at any time. Two patients (both successes) had partial or complete erosion of the collagen sponge. Although the differences were not statistically significant, patients receiving bleomycin had better success rates (8 of 9 versus 6 of 9) and lower intraocular pressure levels (11.88 mmHg versus 14.63 mmHg) than those receiving 5-fluorouracil. The collagen sponge implant demonstrates the clear advantages of drug delivery devices for the administration of antimetabolites after filtration surgery.


Role of magnetic resonance imaging in the evaluation of the hydroxyapatite orbital implant. The role of magnetic resonance imaging (MRI) in the assessment of fibrovascular ingrowth in the integrated hydroxyapatite orbital implant is evaluated. Fifteen patients who underwent enucleation and placement of a hydroxyapatite orbital implant were evaluated for degree of implant vascularity with gadolinium-DPTA-enhanced MRI with surface coil before drilling the implant. On T1-weighted images, the hydroxyapatite sphere appeared with intermediate signal. After gadolinium-DPTA administration, all patients showed an enhancement in the implant consistent with the presence of fibrovascular ingrowth. The enhancement was most notable in the peripheral portions of the sphere and was seen as early as 5 months after implantation. Comparison of gadolinium-DPTA-enhanced MRI with contrast-enhanced computed tomography, ultrasonography, and color Doppler imaging suggests that these latter techniques are not as helpful in the detection of the fibrovascular tissue in the orbital implant. Bone scan, a technique used by many surgeons, demonstrates fibrovascular ingrowth, but it is limited by its one-dimensional low-resolution image. Because of its three-dimensional capability and its highest resolution, contrast-enhanced MRI with surface coil appears to be the best imaging method for evaluating the hydroxyapatite orbital implant and its fibrovascular ingrowth.


Identification of the testis c-mos promoter: specific activity in a seminiferous tubule-derived extract and binding of a testis-specific nuclear factor. The c-mos proto-oncogene is predominantly expressed in male and female germ cells and is involved in the regulation of meiosis. To investigate the mechanism of testis-specific regulation of c-mos transcription, I set out to identify the rat testis c-mos promoter. This was achieved by characterization of the rat testis c-mos transcription start site by primer extension and sequence analysis of cDNAs obtained by polymerase chain reaction amplification of 5' ends of c-mos RNA. The rat testis c-mos transcription start site is located 0.56 kb upstream of the coding region. A fragment containing the rat testis c-mos promoter directs transcription in a nuclear extract derived from rat seminiferous tubules, but not in a liver nuclear extract. DNAase I footprint analysis and gel-retardation assays showed binding of a novel testis-specific nuclear factor to the rat testis c-mos promoter at a site homologous to the testis-specific cis-acting element identified in the promoter of the RT7 gene, which is specifically expressed in haploid male germ cells.


Analysis of phosphotyrosine-containing proteins present in v-src-infected myeloid progenitor cells. We examined the phosphotyrosine-containing proteins in v-src-infected myeloid cells. Proteins with relative molecular weights (M(r)) of 180,000, 175,000, 135,000, 125,000, 120,000, 90,000, 75,000, and 60,000 were present in v-src-infected but not in uninfected 32D cl3 cells. Stimulation of 32D cl3 cells with interleukin 3 (IL-3) resulted in the tyrosine phosphorylation of a protein of 150,000 (M(r)), which was not phosphorylated in v-src-infected 32D cl3 cells. A panel of monoclonal antibodies directed against phosphotyrosine-containing proteins in v-src-transformed chicken embryo fibroblasts (3C4, 2A7, 2B12 and 4F11, directed against p210, p125, p120 and p85 respectively) was used to characterize these substrates. We did not observe tyrosine phosphorylation of proteins recognized by these four monoclonal antibodies in either IL-3-stimulated or v-src-infected 32D cl3 cells. However, we did detect tyrosine phosphorylation of proteins recognized by these monoclonal antibodies in v-src-transformed NIH3T3 cells. Tyrosine phosphorylation of GTPase-activating protein (GAP) and the GAP-associated proteins p62 and p190 was observed in v-src-infected 32D cl3 cells. Stimulation of 32D cl3 cells with IL-3 does not induce phosphorylation of GAP or the GAP-associated proteins p62 or p190. These results suggest that substrates for v-src vary between different cell types.


Angiogenic activity of the K-fgf/hst oncogene in neural transplants. Using retrovirus-mediated gene transfer into neural transplants, we have expressed the human K-fgf/hst oncogene in the central nervous system. Single-cell suspensions of fetal rat brains were removed at embryonic days 13 and 14, exposed to a retroviral vector encoding the K-fgf oncogene and stereotaxically implanted into the caudate putamen of syngenic adult Fisher rats. Recipient animals were sacrificed at intervals of 6-16 months without evidence of neurological impairment. Mock-infected grafts showed the characteristic histopathological appearance of organotypically differentiated neural transplants. In contrast, grafts exposed to the K-fgf gene exhibited abundant capillary proliferation and capillary angiomas. By in situ hybridization analysis and immunohistochemistry, expression of K-fgf was detected in neural cells adjacent to vascular proliferations. Neurons and glia with abundant K-fgf transcripts were morphologically unaffected. In order to examine the transforming potential of the K-fgf gene in the nervous system, we combined retrovirus-mediated transfer of the K-fgf oncogene with a single transplacental exposure of the donor animals to the neurotropic carcinogen N-ethyl-N-nitrosourea (NEU). However, this combination of transforming agents did not result in tumor formation in the grafts. These results provide evidence for a powerful angiogenic effect of K-fgf on the developing brain in vivo.


Cloning and characterization of a thermolabile v-src gene for use in reversible transformation of mammalian cells. The use of temperature-sensitive (ts) src mutants for studies of cell transformation and differentiation has been limited by the availability of cloned ts-src genes that are inactivated at temperatures compatible with growth of mammalian cells. In this report, we describe the cloning and characterization of the tsLA90src gene, which displays tight thermal sensitivity at 39.5 degrees C. Nucleotide sequence comparison of tsLA90 and wild-type src genes from the Schmidt-Ruppin subgroup A and D strains of Rous sarcoma virus (RSV) revealed four amino acid differences in tsLA90src. Substitution of one of these residues (Lys-280) from tsLA90src with its wild-type homolog (Glu-280) caused a reversion to a wild-type src phenotype. The cloned tsLA90 gene, designated tsUP1, was introduced into avian and mammalian retroviral vectors. Chicken embryo fibroblasts and immortalized mouse 3T3 cells infected with these viral vectors displayed a temperature-dependent transformed phenotype as assessed by cell morphology, secretion of plasminogen activator, transcriptional activation of the primary response genes, Egr-1 and TIS 10, and stimulation of tyrosine phosphorylation. In addition, chicken myoblasts (infected with RSVtsUP1) showed a temperature-dependent differentiation into myotubes. Thus, this cloned src gene should be ideally suited for inducing reversible transformation and differentiation of mammalian cells in culture.


Occurrence of distinct PML-RAR-alpha fusion gene isoforms in patients with acute promyelocytic leukemia detected by reverse transcriptase/polymerase chain reaction. A specific 'nested' reverse transcriptase/polymerase chain reaction (RT/PCR) procedure was used to characterize the expression patterns of PML-RAR-alpha chimeric mRNAs in 32 patients with acute promyelocytic leukemia (APL). The sensitivity of the technique was such that the fusion gene transcript could be detected from as little as 2.5 pg of total leukemic cell RNA against a background of 1 microgram of cellular RNA lacking the PML-RAR-alpha fusion gene transcript(s). In 19 cases the PML-RAR-alpha isoform referred to here as long was identified. A short isoform, which in comparison with the long form lacks three PML exons, was detected in 11 other cases. A third PML-RAR-alpha mRNA isoform, in which the most 3' PML exon present in the long-type isoform was truncated in its sequences lying immediately upstream of RAR-alpha B region, was found and characterized in a single patient. In one APL patient with a variant translocation t(11;17), the PCR product corresponding to PML-RAR-alpha chimeric mRNAs could not be amplified despite the presence of RAR-alpha gene rearrangement. Genomic and PCR analysis showed that the different PML-RAR-alpha isoforms found in APL patients arise as a result of distinct translocation breakpoints. In each case the exons encoding the B-F regions of RAR-alpha are expressed and are spliced downstream from variable PML gene exons. The 'nested' RT/PCR analysis of the PML-RAR-alpha fusion gene proved to be a rapid and sensitive tool for the diagnosis of the APL and for monitoring the residual APL chimeric mRNA expression during complete remission.


Functional homology between N-myc and c-myc in murine plasmacytomagenesis: plasmacytoma development in N-myc transgenic mice. Mouse plasmacytomas induced by pristane oil alone, or in combination with Abelson murine leukemia virus (A-MuLV), regularly carry one of three alternative chromosomal translocations that juxtapose c-myc to immunoglobulin heavy- or light-chain loci. E mu-c-myc transgenic mice develop translocation-free plasmacytomas after induction by pristane oil and/or A-MuLV [Sugiyama, H., Silva, S., Wang, Y., Weber, G., Babonits, M., Rosen, A., Wiener, F. & Klein, G. (1990). Int. J. Cancer, 46, 845-852]. In order to test whether another member of the myc family, N-myc, could play a similar role as c-myc, we treated E mu-N-myc transgenic mice with pristane and helper-free A-MuLV. Of 20 mice that received a single pristane injection followed by A-MuLV, 17 developed plasmacytomas with a mean latency period of 54 +/- 20 days. In a corresponding group that only received a single pristane injection, five out of six transgenic mice developed plasmacytomas with a mean latency period of 142 +/- 32 days. However, after three monthly injections of pristane, all 15 transgenic mice developed plasmacytomas with a mean latency period of 128 +/- 20 days. All plasmacytomas expressed the N-myc transgene, while none of them expressed either c-myc or endogenous N-myc. None of the tumors carried the usual plasmacytoma-associated translocations.


Cancer pain relief by continuous administration of epidural morphine in a hospital setting and at home. Fifteen patients with severe pain due to malignancy were treated by continuous epidural morphine infusions. A disposable external pump was used. Patients were treated in a hospital setting or at home for a total of 906 days. Pain intensity was estimated by VAS. The pumps functioned well. Bacterial growth was found in 0.6% of the balloon reservoirs used, while the epidural filters were free from growth. There were no clinical infections. It appears that this delivery system is safe, practical and suitable for use in the home environment.


Organic hallucinosis in patients receiving high doses of opiates for cancer pain. We report 4 patients who developed organic hallucinosis while receiving opiate analgesics for cancer pain. In all patients, the symptoms rapidly responded to haloperidol and change in the type of opiate. The hallucinations were exclusively visual. Cognitive status, determined by the Mini-mental State Questionnaire (MMSQ), was normal in all patients. Organic hallucinosis occasionally occurs in patients receiving opiates and can be a major source of distress for patients and their families.


Relationship between autotomy behaviour and spinal cord monoaminergic levels in rats. In the rat, unilateral neurectomy of the sciatic and saphenous nerves causes autotomy, a self-mutilation behaviour, against the denervated limb that is variable in both its onset and severity. To study some of the possible neurochemical sources of this variability, spinal cord levels of norepinephrine (NE), dopamine (DA), serotonin (5-HT0 and 5-hydroxyindoleacetic acid (5-HIAA) were analysed ipsi- and contralateral to the lesioned side by high performance liquid chromatography at C5-T1 and L1-S1. According to the early or late onset and to the slight or intense autotomy behaviour, the animals were assigned to four different groups: early autotomy, early no autotomy, late autotomy, and late no autotomy. Two sham-operated groups were sacrificed at an early or late stage in the postoperative period. The spinal cord NE content remained unchanged throughout the different experimental situations. The more conspicuous changes observed were: (1) a generalized increase in spinal 5-HT metabolism in all deafferented groups; (2) a significant and selective increase in lumbosacral 5-HT and 5-HIAA levels of the rats that did not self-lesion for 8 weeks after deafferentation and (3) a significant fall (30-45%) in DA levels at denervated spinal segments of the rats that actively self-attacked late in the postoperative period. The data suggests that spinal cord serotonergic and dopaminergic influences play an important role in determining the susceptibility to autotomy (and perhaps chronic pain) after peripheral deafferentation.


Do opioids evoke the release of serotonin in the spinal cord? An in vivo microdialysis study of the regulation of extracellular serotonin in the rat. This study investigated the regulation of serotonin (5-HT) and its major metabolite 5-hydroxyindoleacetic acid (5-HIAA) in the dorsal spinal cord of awake, freely moving rats, using microdialysis coupled to HPLC with electrochemical detection and tested the hypothesis that opioids exert their analgesic effect in part through the increased release of 5-HT in the dorsal horn. A dialysis tube was placed transversely at the L4 segment of the dorsal spinal cord and the basal concentration of 5-HT in the dialysate was characterized by infusion of a variety of substances through the dialysis probe: tetrodotoxin (TTX), KCl, imipramine, fluoxetine and amphetamine (AMPH). To evaluate the contribution of opioids, we also studied the effects of either systemic or intracerebroventricular (i.c.v.) injection of morphine or DAMGO. Extracellular concentrations of 5-HT and 5-HIAA were partially and reversibly reduced by TTX. In the presence of KCl, imipramine, fluoxetine or AMPH, 5-HT levels significantly increased. Under these conditions, extracellular 5-HIAA levels usually decreased. By contrast, the effects of opioids on 5-HT concentrations were highly variable. Low doses of morphine administered systemically increased 5-HT concentrations in only 3 of 6 rats. This was paralleled by a decrease in 5-HIAA. Higher doses of morphine, alone or in the presence of fluoxetine, did not change 5-HT concentrations. Intracerebroventricular injection of morphine or DAMGO increased the extracellular concentrations of 5-HT in only about one third of the animals. After intracerebroventricular opioid injection, extracellular concentrations of 5-HIAA either decreased by about 20% or did not change.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of exposure to sauna heat on neuropathic and rheumatoid pain. Exposure to sauna heat during sauna bathing raises the skin temperature of the bather near the hot pain perception threshold and enhances sympathetic activity. Self-reports provided by regular bathers of changes in intensity of their ongoing pain might, therefore, add novel information on the effect of intense heat on various pain conditions. We interviewed consecutive patients attending our pain clinic over a period of 1 year about their pain-related responses to sauna bathing and controlled the results by quantitated somatosensory tests. There were 61 patients with chronic neuropathic pain of peripheral origin, 13 patients with central pain and 59 patients with rheumatoid pain. Allodynia and hyperalgesia to heat were relatively infrequent in all groups (10%, 15% and 8%, respectively). Three out of 17 patients with postinjury nerve pain reported similar exacerbation. By contrast, mechanical allodynia was present in 48% of patients with peripheral neuropathic pain and in 54% of patients with central pain. The results speak against an important role for C-afferent or sympathetic postganglionic fibres in most subclasses of neuropathic pain. Animal models of neuropathic pain should be critically viewed against this finding.


Individual variability in the response to different opioids: report of five cases. Although it is widely appreciated that patients can demonstrate highly variable responses to different opioid drugs, there have been few detailed descriptions of this phenomenon. To illustrate this variability, we present 5 patients, 4 with cancer pain and 1 with non-malignant pain, who underwent dose titration with more than 1 opioid and developed markedly different responses to each. In every case, dose escalation led to successful treatment with 1 opioid and to intolerable side effects without adequate relief with others. The existence of this individual variability in the response to different opioids has important implications for both clinical practice and current understanding of opioid pharmacology in man. It contradicts the view that any opioid is inherently more efficacious than any other, suggests that patients who fail to obtain adequate pain relief at maximally tolerated doses of 1 opioid may benefit from an alternative drug, and underscores the potential importance of genetic factors as a determinant of opioid response.


Tumor necrosis factor-alpha in human milk. We previously demonstrated that certain biologic activities in human milk were partially blocked by antibodies directed against human tumor necrosis factor-alpha (TNF-alpha). In this study, immunochemical methods were used to verify the presence of TNF-alpha in human milk obtained during the first few days of lactation. Gel filtration revealed the presence of TNF-alpha by RIA in molecular weight fractions between 80 and 195 kD. TNF-alpha could not be detected consistently by conventional Western blotting or cytotoxic assays. Although immunoreactive bands were detected by a Western blot-125I protein A technique in TNF-alpha-positive fractions from gel filtration, those bands proved to be nonspecific. TNF-alpha in milk was reliably quantified by the competitive RIA. Those studies revealed that the concentrations of TNF-alpha in milk were 620 +/- 183 pg/mL. Although RNA to TNF-alpha was detected in milk leukocytes by Northern blotting, little TNF-alpha was found in those cells before or after stimulation with N-formyl-l-methionyl-l-leucyl-l-phenylalanine or 4 beta-phorbol-12 beta-myristate-13 alpha-acetate. The origin of this cytokine in human milk remains unclear. Nevertheless, this study suggests that TNF-alpha is present in early human milk in sufficient quantities to exert possible biologic effects upon the mammary gland of the mother or the immune system of the infant.


Increased red cell aggregation does not reduce uteroplacental blood flow in the awake, hemoconcentrated, late-pregnant guinea pig. The effect of increased red blood cell aggregation on uteroplacental blood flow was studied in 11 awake, late-pregnant guinea pigs. The aggregation of the red cells was increased by administering high molecular weight dextran (HMWD) to the previously hemoconcentrated animal. The purpose of the hemoconcentration before HMWD was 1) to use a preeclampsia model in which the hemorheology may be impaired because of the combined effect of polycythemia, an increased red cell aggregation, and an increased plasma viscosity and 2) to potentiate the aggregation-increasing effect of HMWD. Relative to the pre-HMWD condition, arterial blood pressure and systemic vascular resistance increased by 10 and 26%, respectively. The cardiac output fraction shunted across the systemic circulation and the arterial hematocrit decreased by 30 and 4%, respectively. Neither cardiac output nor the weighted organ flows, including those to the placentas, changed in response to the rise in red cell aggregation. We conclude that an imposed increase in red cell aggregation has no appreciable effect on uteroplacental blood flow in the awake and healthy late-pregnant guinea pig. These data do not exclude the possibility that increased red blood cell aggregation potentiates the negative effects on uteroplacental blood flow, e.g. in pregnancy-induced hypertension or preeclampsia, where the placenta is not only marginally perfused but also frequently damaged histologically.


Enhancing the cognitive outcomes of low birth weight, premature infants: for whom is the intervention most effective? The Infant Health and Development Program is a national collaborative study to test the efficacy of combining early child development and family support services with pediatric follow-up to reduce the incidence of health and developmental problems among low birth weight, preterm infants in eight medical school sites. Its efficacy in enhancing intellectual outcomes at age 3 in more and less environmentally vulnerable, low birth weight, preterm children, as defined by maternal education (high school completion or less vs some college) and race (black vs white/other), is explored. Children whose mothers had a high school education or less benefited from the intervention. This was true for both the black and white samples. Children whose mothers had attended college did not exhibit significant enhancement in IQ scores at 3 years. Birth weight affected the response to treatment for one of the four subgroups: Among white mothers with some college, the lighter (less than 2000 g) low birth weight, preterm children were less influenced by the intervention than were the corresponding heavier children. Implications for targeting certain subgroups of low birth weight, preterm children for services are considered.


Accuracy of the Denver-II in developmental screening. One of the oldest and best known developmental screening tests was recently restandardized and revised as the Denver-II. Because it was published without evidence of its accuracy, the present study was undertaken with 104 children between 3 and 72 months of age attending one of five day-care centers. To determine the presence of developmental problems, children were administered individual measures of intelligence, speech-language, achievement, and adaptive behavior. A second psychological examiner, blind to the outcome of the diagnostic battery, administered the Denver-II. Developmental problems including language impairments, learning disabilities, mild mental retardation, and/or functional developmental delay were found in 17% of the children. The Denver-II identified correctly 83% and thus had high rates of sensitivity. However, more than half the children with normal development also received abnormal, questionable, or untestable Denver-II scores. Thus the test had limited specificity (43%) and a high overreferral rate. The alternative scoring method, categorizing questionable/untestable scores as normal, caused sensitivity to drop to 56% although specificity rose to 80%. Since neither scoring method produced acceptable levels of accuracy, an effort was made to locate the sources of accuracy and inaccuracy within the test. Only items in the language domain were modestly helpful in discriminating children with and without difficulties. The findings suggest that the authors of the Denver-II need to engage in further development of the instrument including revising scoring criteria and item placement in relation to children's ages. In the interim, test users should employ screening tests which are more accurate such as the Minnesota Inventories or the Battelle Developmental Inventory Screening Test.


The T-loop region of animal mitochondrial tRNA(Ser)(AGY) is a main recognition site for homologous seryl-tRNA synthetase. Recognition sites of bovine mitochondrial serine tRNA specific for condons AGY [tRNA(Ser) (AGY)] by the cognate mitochondrial seryl-tRNA synthetase were studied using a range of tRNA(Ser)(AGY) variants which were obtained by the in vitro transcription of synthetic tRNA genes with T7 RNA polymerase. Base replacements in the anticodon and discriminator sites did not affect serine acceptance. However, deletion and/or replacement in the T-loop region completely deprived the variants of their charging activities. Point mutation experiments in this region also showed that the adenosine residue in the middle of the T-loop (position 58), which is involved in tertiary interaction between the T-loop and the truncated D-arm [de Bruijn and Klug, 1983] played a significant role in the recognition process by the synthetase.


In vitro study of E.coli tRNA(Arg) and tRNA(Lys) identity elements. Various tRNA transcripts were constructed to study the identity elements of E.coli tRNA(Arg) and tRNA(Lys). Exchange of the anticodon of the major tRNA(Arg) from ACG to either CCG or CCU did not result in a significant loss of arginine acceptor activity, whereas not only that to UUU but also that to ACA or ACC decreased the activity. Base substitutions and deletion at A20 also impaired the arginine charging activity by over 50-fold. Arginine charging activity was introduced by either substitution of the anticodon from UAC to ACG in tRNA(Val) or from UUU to UCU in tRNA(Lys). Only a single base substitution at the third position of tRNA(Trp) anticodon (CCA) from A to G also gave rise to arginine charging activity, which was elevated to a comparable level to that of the tRNA(Arg) transcript by an additional A20 insertion. Base substitutions of the major tRNA(Arg) at the discriminator position into pyrimidines led to a decrease by factors of three to four. These data show that the third letter of the anticodon G36 or U36 besides the second letter C35 and the A20 in the variable pocket is responsible for the arginine acceptor identity, to which the discriminator base A73 or G73 contributes in an auxiliary fashion. In contrast to the arginine system, the transcript with the wild-type tRNA(Lys) sequence showed only 140-fold lower lysine charging activity than the native tRNA(Lys), suggesting the involvement of base modifications in recognition. Replacement of the anticodon UUU with not only UCU and UAC but also UUA and UUC seriously affected the lysine acceptor activity, and those with GUU and UUG also decreased by factors of 17 and 5, respectively. Introduction of UUU into the anticodons conferred lysine charging activity upon both tRNA(Val) and tRNA(Arg). Substitution of the discriminator base A73 by any of the other bases decreased the lysine acceptor activity by a factor of ten. These results indicate the involvements of all the three bases of the anticodon and A at the discriminator position in lysine specific aminoacylation.


Mutational evidence for competition between the P1 and the P10 helices of a mitochondrial group I intron. A guanosine to cytosine transversion at position 2 of the fifth intron of the mitochondrial gene COB blocks the ligation step of splicing. This mutation prevents the formation of a base pair within the P1 helix of this group I intron--the RNA duplex formed between the 3' end of the upstream exon and the internal guide sequence. The mutation also reduces the rate of the first step of splicing (guanosine addition at the 5' splice junction) while stimulating hydrolysis at the 3' intron-exon boundary. Consequently, the ligation of exons is blocked because the 3' exon is removed prior to cleavage at the 5' splice junction. The lesion can be suppressed by second-site mutations that preserve the potential for base-pairing at this position. Because the P1 duplex and the P10 duplex (between the guide sequence and the 3' exon) overlap at the affected pairings represent alternative structures that do not, indeed cannot, form simultaneously.


Flow cytometry: an overview.

Flow cytometry: an overview. Flow cytometry is a method of rapidly analyzing large numbers of cells as they flow in a liquid medium through a laser source. Flow cytometry provides valuable information regarding DNA content, RNA content, and the percentage of cells in the S phase of the cell cycle. This information can be used to predict the aggressiveness of many tumors, which has direct prognostic and diagnostic implications. Flow cytometry assists healthcare professionals in planning and implementing effective, appropriate, and timely interventions. Because flow cytometry is becoming more prevalent, nurses must be aware of its actual and potential applications. The data obtained from flow cytometry provide important information that can assist nurses in anticipating a particular regimen for specific tumors and in projecting the patient's clinical course. With this information, nurses can modify and individualize care plans. When used in conjunction with a patient's clinical presentation and other laboratory findings, flow cytometry has a direct impact on both treatment decisions and nursing interventions.


Cardiac myxoma with a cytokeratin-immunoreactive glandular component. This study documents the expression of cytokeratin intermediate filaments (IFs) in a surgically excised glandular atrial myxoma. The glandular structures showed also positivity for carcinoembryonic antigen. Typical isolated or cordlike myxoma cells contained vimentin IFs. Positivity for von Willebrand factor was detected in cells lining deep invaginations of surface papillae and vascular channels. Bundles of smooth muscle cells and myofibroblasts scattered throughout the myxoid matrix synthesized the isoform of alpha-actin specific for smooth muscle.


A monoclonal antibody that recognizes a formalin-resistant epitope on the p 24 core protein of HIV-1. A murine monoclonal antibody (called H-11) that binds to the p 24 core protein of HIV-1 was characterized by radioimmuno-precipitation, immunofluorescence, western blot assays, immunocytochemistry and immunohistochemistry. This antibody was found to be especially suited for demonstrating the presence of HIV-1 in formalin-fixed, paraffin-embedded tissues.


Correlated responses in lines of chickens divergently selected for fifty-six-day body weight. 2. Organ growth, deoxyribonucleic acid, ribonucleic acid, and protein content. Growth of organs relative to body weight and cellular protein, RNA, DNA, and cell unit size of breast muscle, liver, and small intestinal tissue were measured in females from four lines of chickens. Two lines had undergone 32 generations of divergent selection for 56-day body weight, and the other two lines were derived by sampling the first two lines at Generation 28 and relaxing selection for the next five generations. The diet used in the present experiment was the same diet under which selection was practiced (20% crude protein and 2,685 kcal of ME/kg). Comparisons at common chronological ages and a common body weight revealed that supply organ weights, especially that of the small intestine, were associated with subsequent growth of demand organs. Although the upper gastrointestinal tract was also important in this respect, it was more susceptible to influences such as feed intake. Selection for juvenile body weight resulted in correlated changes in cell size of breast muscle but not liver and small intestine. Muscle increased posthatch as cells underwent hypertrophy but liver and small intestine grew chiefly by hyperplasia.


Correlated responses in lines of chickens divergently selected for fifty-six-day body weight. 3. Digestive enzymes. Levels of amylase, trypsin, chymotrypsin, and lipase in the pancreas and small intestinal chyme were measured in females from four lines of chickens. Two of the lines had undergone 32 generations of divergent selection for 56-day body weight, and in the other two lines selection for high or low weight had been relaxed for 5 generations. The diet used in the present experiment was that under which selection had been practiced (20% crude protein and 2,685 kcal of ME/kg). Comparisons between divergently selected lines at common ages revealed higher enzyme levels for high- than low-weight lines. When comparisons were made at a common body weight (80 +/- 5 g) there were no differences between lines. These results suggested that correlated responses in feed intake were mediating the regulation of digestive enzyme levels in the pancreas and in intestinal chyme of growth-selected lines of chickens. Chicks from high-weight lines had elevated enzyme levels after a mild feed restriction compared with those provided ad libitum access to feed. It was hypothesized that hyperphagia associated with the high-weight lines in combination with a mild feed restriction and the associated meal feeding stimulated synthesis and secretion of digestive enzymes.


Plasma cells expressing immunoglobulins M and A but not immunoglobulin G develop an intimate relationship with central canal epithelium in the harderian gland of the chicken. In the Harderian gland of the chicken, the epithelial and plasma cell relationships were studied by light and electron microscopy and immunohistochemical methods. In the wall of the central canal a dark epithelial cell was identified that had long branching cell processes. An anticytokeratin monoclonal antibody demonstrated that the dark cells provided an extremely large contact area for the plasma cells. Although IgM-, IgG-, and IgA-producing plasma cells were present in the Harderian gland, only IgM- and IgA-positive cells were capable of a distinct relationship with dark epithelial cells. The surface of the primary branches contained scattered IgA deposits whereas the epithelial cells of the secondary branches possessed IgA along the lateral cell membrane but not on the surface. Anti-IgA and anti-cytokeratin antibodies produced a similar staining pattern in the acini and secondary branches. Taken together, these observations suggest that IgA secretion is a function of secondary branches and that intracellular transport is influenced by the cytoskeletal system.


Generation and possible significance of trypsinogen activation peptides in experimental acute pancreatitis in the rat. Trypsinogen activation peptides (TAP) were quantified by radioimmunoassay in blood, urine, and peritoneal exudate of rats with experimental pancreatitis. Forty-four animals were studied, comprising a control group and four different induction techniques (cerulein, cerulein plus either 2- or 10-min intraductal glycodeoxycholic acid [GDOC] infusion, and cerulein plus intraductal GDOC with enterokinase [EK]). Significantly higher TAP concentrations were found at 6 h (or at death) in plasma and ascites of all pancreatitis groups compared with controls. TAP quantitation in hourly urine samples demonstrated significantly higher concentrations from the third hour onward in the most severe groups and from the fourth hour onward in the cerulein-treated rats. All nonsurviving rats had a plasma TAP of greater than 2.5 nM/L, whereas only 1 of 34 surviving animals had such a concentration (p less than 0.001). A significant stepwise increase in total TAP in ascites was found when comparing the cerulein group, the two GDOC groups, and the EK group (p less than 0.001). Chromatography of samples with a high TAP content demonstrated comigration with synthetic TAP. We conclude that free TAP are present in blood, urine, and peritoneal exudate of rats with experimental pancreatitis of different pathogenesis and that the amount of TAP may be indicative of the severity of the disease process.


Endocrine and exocrine function of pancreatic fragments autotransplanted into hepatic parenchyma. We studied the use of hepatic parenchyma as a recipient site for pancreatic fragment transplantation. Endocrine and exocrine pancreatic functions were evaluated following pancreatic autotransplantation in 26 mongrel dogs that had undergone total pancreatectomy. The endocrine function of the pancreatic tissue transplanted to hepatic parenchyma was significantly inferior to that of normal controls. Cholecystic bile amylase concentrations were markedly elevated in six dogs that had been implanted with pancreatic fragments in their hepatic parenchyma and had survived more than 2 months. Also, choledochal bile amylase concentrations increased significantly following pancreozymin-secretion (PS) injection. In contrast, cholecystic bile amylase concentrations in normal dogs were low and choledochal bile amylase concentrations did not respond to a PS load. Histological examination of pancreatic autografts in hepatic parenchyma revealed marked proliferation of exocrine tissue with abundant zymogen granules and reconstruction of pancreatic lobules with a few islets.


Formation of pseudoislets from human pancreatic cultures. We have successfully developed a technique for culturing human islet cells obtained from the cadaveric pancreata of children. Within 24-48 h of in vitro culture, collagenase-digested human pancreatic tissue formed epithelioid monolayers. Scattered within these monolayers were insulin-positive cells, as detected by immunocytochemical and dithizone staining. Treatment of the beta cell-containing epithelioid-cell monolayers with EDTA resulted in the formation of spherical cellular clusters, i.e., pseudoislets. These pseudoislets differed from isolated islets of Langerhans in that they showed a more peripheral distribution of insulin-positive cells. Our studies have demonstrated that insulin-positive cells can be detected in monolayers obtained from human pancreata 3-4 weeks after culture. When exposed to varying concentrations of glucose, these cells secreted insulin. The development of this in vitro technique for culturing human pancreatic islet tissue could provide a model for systematically studying in vitro islet function.


[Interaction of ovomucoid from duck egg proteins with aldehydes-dextrans]. The interaction of ovomucoid proteinase inhibitor prepared from duck egg white with a dextran of a molecular weight of 70,000 preliminary treated with potassium periodate. Irrespective of the number of the sites of the ovomucoid binding to aldehyde-dextran the anti-chymotryptic activity is equal to that of the native inhibitor, while the antitryptic activity decreases proportionally to the number of ovomucoid amino groups involved in the reaction with dextran. When a few ovomucoid molecules are immobilized on the polysaccharide macromolecule the perturbing effect of the protein-protein interactions is minimal, as the rigid polymeric chain prevents from the formation of associates of proteins immobilized on this backbone.


A preliminary 3-D model of the tertiary fold of the polymerase domain of HIV-1 reverse transcriptase. Development of a 3-D model of the reverse transcriptase from type 1 human immunodeficiency virus (HIV-1 RT), a key enzyme in the pathogenesis of the virus, presents a significant challenge. Three-dimensional structural information is not available for any close homolog, the only 3-D structural data being that of the Klenow fragment (KF) of Escherichia coli DNA polymerase I, for which coordinates of only the alpha-carbons are available. A recently published study of the sequences of a large number of polymerases led to the identification of three common sequence patterns, nominally motif A, motif B and motif C, and to the hypothesis that the various DNA and RNA polymerases including E. coli DNA polymerase I and HIV-1 RT share a common structural motif around their respective polymerase active sites. The preliminary results of recent structural studies on two other polymerases also support this hypothesis. Based on the assumption of structural homology in the active site regions of their polymerase domains, the HIV-1 RT and KF sequences were aligned using pattern-based secondary structure predictions as a guide and motifs A, B and C as 'anchor points'. However, as suggested by the results of chemical modification experiments, it was assumed that the order of the motifs in KF, viz. A, B and C, differed from that of the related motifs A, C and B' in HIV-1 RT, a rearrangement that could have been brought about by an exon shuffling type of mechanism.(ABSTRACT TRUNCATED AT 250 WORDS)


Heat capacity changes and hydrophobic interactions in the binding of FK506 and rapamycin to the FK506 binding protein. Differential interactions among nonpolar moieties at protein/ligand interfaces, and of these nonpolar groups with water, collectively termed hydrophobic interactions, are widely believed to make important energetic contributions to the stability of protein/ligand complexes. Quantitative estimates of hydrophobic interactions, and an evaluation of their structural basis, are essential for obtaining structure-based predictions of the free energies of binding for the purpose of drug design. Two largely nonpolar, immunosuppressive agents, FK506 and rapamycin, each bind with high affinity to a common hydrophobic pocket on a small peptidylproline cis-trans isomerase known as FK506 binding protein (FKBP-12) and inhibit its activity. In an effort to elucidate the structural features of these ligands responsible for the observed energetics, we have undertaken an investigation of the thermodynamics of binding of FK506 and rapamycin to FKBP-12. Enthalpies of binding have been determined by high-precision titration calorimetry over a range of temperature, allowing estimates of heat capacity changes. By analyzing the distribution of changes in solvent-accessible surface area upon binding of FK506 to FKBP-12 from crystallographic data, it is found that 99% of the net surface buried upon binding involves nonpolar groups. This leads to a heat capacity change of FK506 binding, normalized to the amount of nonpolar surface, of -0.40 +/- 0.02 cal.K-1.mol-1.A-2 (1 cal = 4.18 J), a value similar to that obtained for the aqueous dissolution of hydrophobic substances. Our observations are discussed in view of the general nature of hydrophobic interaction processes.


Purified unitary kainate/alpha-amino-3-hydroxy-5-methylisooxazole-propionate (AMPA) and kainate/AMPA/N-methyl-D-aspartate receptors with interchangeable subunits. We have purified and characterized two vertebrate excitatory amino acid ionotropic receptors from the Xenopus central nervous system. Each is a unitary receptor (i.e., having more than one class of excitatory amino acid agonist specificity within one protein oligomer). The first is a unitary non-N-methyl-D-aspartate (non-NMDA) receptor and the second is a unitary NMDA/non-NMDA receptor. The specific agonist-activated channel activity and pharmacology of each type were recognized by patch-clamping lipid bilayers in which the isolated protein was reconstituted. In the second case, the NMDA and the non-NMDA sites could not be physically separated and exhibited functional interaction. Parallel evidence for this was obtained when poly(A) RNA from Xenopus brain was translated in oocytes: a noncompetitive inhibition of the response to L-kainate is produced by NMDA to a maximum depression of 30% at 1 mM NMDA. Each isolated oligomer contains 42-kDa subunits of the non-NMDA ligand binding type, but the second type has an additional NMDA-receptor-specific 100-kDa subunit. Thus, a subunit-exchange hypothesis can account for the known multiplicity of excitatory amino acid receptor types.


Translocation of spectrin and protein kinase C to a cytoplasmic aggregate upon lymphocyte activation. We have previously reported that mammalian tissue lymphocytes exhibit significant heterogeneity with respect to the subcellular distribution of spectrin and that this phenomenon may result from a dynamic behavior of spectrin in response to activation signals. Here, we further characterize the involvement of spectrin in lymphocyte activation by examining its relationship with protein kinase C (PKC). PKC isoenzymes are a family of cytosolic kinases that translocate from the soluble to particulate fraction upon cell stimulation. It is reported here that activation of lymph node T cells through the antigen-specific receptor, or direct activation of PKC by phorbol esters, results in a striking increase in cells expressing a cytoplasmic aggregate of spectrin. Additionally, a concurrent increase in cells expressing aggregates of the beta II isozyme of PKC is observed. Immunofluorescence staining revealed that spectrin and PKC beta II are colocalized in untreated lymphocytes and that these two proteins are coincidently translocated to the same focal aggregate within the cytoplasm following stimulation. This redistribution of spectrin and PKC beta is blocked by pretreatment with calphostin C, a specific inhibitor of PKC. Solubility studies showed that there is an increase of both proteins in the detergent-insoluble fraction of lymphocytes upon activation, and immunoprecipitation studies indicated that the soluble form of these molecules may be associated directly or indirectly as part of a complex of proteins. These data indicate that the positioning of the spectrin-based cytoskeleton is sensitive to activation signals and may play a role in the function or positioning of PKC beta II.


Two types of anion channel currents in guard cells with distinct voltage regulation. Transpirational water loss by plants is reduced by closing of stomatal pores in the leaf epidermis. Anion channels in the plasma membrane of guard cells may provide a key molecular mechanism for control of stomatal closing in leaves. However, central questions regarding the regulation, diversity, and function of anion channels in guard cells and other higher plant cells remain unanswered. We show here that two highly distinct types of depolarization-activated anion currents operate in the plasma membrane of Vicia faba guard cells. One described type of anion channel was activated rapidly within 50 ms by depolarization, inactivated during prolonged stimulation, and deactivated rapidly at hyperpolarized potentials (R-type anion current). The other depolarization-activated anion current showed extremely slow voltage-dependent activation and deactivation (S-type anion current) and lacked inactivation. The distinct voltage and time dependencies of R-type and S-type anion channels suggest that they may play a role during depolarization-associated signal transduction in higher plant cells and that these anion channels may contribute to different processes in the regulation of stomatal movements. In particular, the slow and sustained nature of S-type anion channel activation revealed here leads us to hypothesize that S-type anion channels may provide a central molecular mechanism for control of stomatal closing, which is accompanied by long-term anion efflux and depolarization.


Higher order interactions in 23s rRNA.

Higher order interactions in 23s rRNA. An alignment of 75 phylogenetically diverse large subunit ribosomal RNA sequences was created and searched for secondary and tertiary structure elements by computer. The search revealed four unknown secondary structural pairings, two internal loop closings, and five short-range tertiary interactions--two of which were pairings of an unusual type. One brings a loop together with two other loops previously known to be paired, and one involves a nucleotide within a presumed tetraloop. The latter interaction constrains the RNA structure near the ribosomal E-site, where two base pairs previously suggested to be in parallel orientation are now proven. No clear phylogenetic evidence for direct base pairing between the large and small subunit rRNA was found.


Interleukin 6: insights to its function in skin by overexpression in transgenic mice. Interleukin 6 (IL-6) is a cytokine that mediates a wide range of inflammatory and immune responses. Its expression is elevated in inflammatory or immunodeficient diseases, including psoriasis, rheumatoid arthritis, and AIDS. To explore the role of IL-6 in skin, we utilized a human keratin 14 (K14) promoter to express IL-6 in the basal cells of stratified squamous epithelia of transgenic mice. Mice expressing the K14-IL-6 transgene were smaller than normal and exhibited retarded hair growth. Surprisingly, IL-6 expression did not lead to enhanced epidermal proliferation, but it did result in a thicker stratum corneum, with an otherwise seemingly normal program of differentiation. IL-6 expression did not lead to leukocytic infiltration, making it unlikely that it has direct proinflammatory activity in skin. Based on this study, one role of IL-6 relevant to host defense may be to enhance the stratum corneum, thereby providing increased protection from injurious stimuli or infection. If IL-6 plays additional roles in the skin, it is likely to act synergistically with factors that IL-6 alone cannot induce.


Spinal neurons exhibiting a specific nociceptive response receive abundant substance P-containing synaptic contacts. Substance P has been implicated in nociceptive transmission in the spinal cord. However, evidence for a direct correlation between a specific nociceptive response in spinal dorsal horn neurons and substance P input is lacking. In this study, we combine intracellular recording from dorsal horn neurons in vivo, characterization of their nociceptive responses, intracellular labeling by injection of horseradish peroxidase, and immunocytochemical demonstration of substance P at the electron microscopic level. The results reveal that dorsal horn neurons that respond to noxious cutaneous stimulation with a slow, prolonged excitatory postsynaptic potential receive a preferentially high number of substance P fibers compared with nonnociceptive neurons, which scarcely receive any substance P input. Therefore, this study provides direct evidence of a structural-functional link for a substance P-mediated nociceptive response.


Urinary indoleamines in Tourette syndrome patients with obsessive-compulsive characteristics. Tourette syndrome patients with high levels of obsessive-compulsive symptoms were compared with patients without these symptoms on urinary measures of serotonin and its major metabolite, 5-hydroxyindoleacetic acid (5HIAA). Both groups were compared with normal controls, and it was hypothesized that patients with obsessive-compulsive symptoms would have lower levels of serotonin. Both groups of Tourette syndrome patients had lower levels than controls, but there was no difference between them. Obsessive symptoms were related to higher levels of 5HIAA and to a higher turnover of serotonin.


Characterization of immortal cystic fibrosis tracheobronchial gland epithelial cells. Tracheobronchial glands were isolated and cultured from a patient with cystic fibrosis (CF). Cultured epithelial cells were transformed with pSVori-. All transformed cell lines express cytokeratin filaments and at early passages express the junctional complex molecule cell CAM 120/80, indicating their epithelial origin. Several gland cell lines express antigens that localize to secretory cells in vivo. Cl- transport measured by 36Cl efflux shows that CF gland epithelial cells, like CF surface airway and nasal polyp epithelial cells, are unable to respond to increases in intracellular cAMP. However, they do produce an increase in intracellular cAMP after treatment with isoproterenol or forskolin. One CF gland cell line shows increased intracellular calcium in response to a number of agents and increased Cl- efflux comparable to that observed in a non-CF airway surface epithelial cell line after addition of calcium ionophore. All cell lines express CF transmembrane conductance regulator mRNA, as measured by PCR amplification of first-strand cDNA. The CF tracheobronchial gland cell lines described here are compound heterozygotes, having a single copy of the delta F508 mutation.


Feasibility of interactive videodisc technology to teach minority youth about preventing HIV infection. Hispanic and African American adolescents are more likely than white Anglo youth to harbor misconceptions about acquired immunodeficiency syndrome (AIDS) and are also more likely to engage in intravenous drug use and sexual intercourse. This paper describes the development of an AIDS prevention curriculum that uses an interactive videodisc program to teach skills for interventions. Focus group and expert panel studies yielded suggests for intervention vignettes and scenes relevant to Hispanic and African American adolescents. The authors then developed and produced a sample curriculum, specifically designed for Hispanic youth. Content was designed to build knowledge, attitudes, and skills in handling situations where young persons are at risk for human immunodeficiency virus (HIV) infection. The feasibility of the finished pilot product was tested with adolescents and with professionals who serve ethnic and racial minority youth. Adults and Hispanic adolescent viewers rated the videodisc as enjoyable, interesting, and likely to achieve positive effects with the intended target population. Findings suggest that the interactive videodisc is a useful way to interest and help Hispanic adolescents learn ways of reducing their risk of contracting and spreading HIV infection through lifestyle practices. This developmental research in the use of interactive videodisc also provides a basis for further investigation.


Cisplatinum-induced lesion of proximal tubule acidification in the rat. Rats were given a 4- to 6-mg/kg body weight intraperitoneal injection of the antitumor drug, Cisplatin, 5-7 days prior to experiments to study tubule acidification by clearance and stationary microperfusion techniques. Cisplatin reduced the glomerular filtration rate markedly and caused a moderate degree of metabolic acidosis, but urine acidification (pH) was well maintained. Proximal tubule stationary pH and bicarbonate concentrations, as measured by pH microelectrodes, were significantly increased. The defect of proximal H+ secretion is reflected by increased acidification half-times (from 4.44 to 10.2 s) and reduced bicarbonate reabsorption to 37% of control values. H-ion back flux, measured during tubule and capillary perfusions with Ringer's bicarbonate- and CO2-free phosphate solutions, was reduced to 68% of control values. The apparent H-ion permeability was lowered from 0.79 to 0.54 cm/s. These results indicate that proximal acidification is reduced by impairment of H+ transport and not by increased transepithelial H+ shunting. Blunted acidification is compatible with a reduction in the number of Na/H exchangers in the proximal brush border and/or a decrease in the apical sodium gradient, the driving force for proximal H-ion secretion. Cortical distal tubule acidification, measured by double-barreled ion-exchange resin/PD microelectrodes, was not significantly affected by Cisplatin. This accounts for the observation that, in spite of the impaired proximal acidification, urine pH is kept within the normal range.


Desensitization to vasopressin action in the rat kidney medulla: studies on isolated nephron segments. Because of the prominent role of the renal medulla in the elaboration of concentrated urine and the possible differential regulation of the various nephron segments, desensitization to vasopressin (AVP) was studied on freshly isolated rat medullary thick ascending limbs (MTALs) and outer medullary collecting ducts (MCDs). Desensitization was induced through intramuscular injections of 1-deamino-8-D-arginine-vasopressin (dDAVP, 2 micrograms/day for 3 days), the last of which was performed 1 h before kidney removal. The cellular response to AVP was studied by measuring cAMP accumulation in micro-dissected nephron segments. In the MTAL, we observed a marked (around 80%) and selective desensitization to AVP, the response to either glucagon or human calcitonin remaining unaltered. In the MCD, significant desensitization was observed in the presence of 1 nM AVP in the assay medium, and was no longer significant when the AVP concentration was increased to supramaximal levels (10-1,000 nM). These experiments thus reveal a clear dissociation between MTAL and MCD with regards to dDAVP-induced desensitization and extend previous observations made on target segments in the renal cortex.


[Immunotherapy of recurrent spontaneous miscarriages (idiopathic abortive disease): preliminary results]. Recurrent spontaneous abortion (greater than or equal to 3 spontaneous miscarriages) represents an entity defined by negative criteria (absence of anatomical, hormonal, autoimmune and chromosomal abnormalities). The immune hypothesis is corroborated by the successes (greater than or equal to 80 %) of specific (paternal leucocytes) or non-specific (intravenous gammaglobulins) immunotherapeutic trials. Studies on the mechanisms of action of those two methods will afford information on the pathogenesis of this condition.


[Prostate carcinoma--a current review].

[Prostate carcinoma--a current review]. Carcinoma of the prostate is the most commonly diagnosed cancer in men. The natural history and the biological aggressiveness are primarily determined by tumor volume. At the time of diagnosis, only one third of all tumors are pathologically confined to the prostate and eligible for curative therapy. Early detection by the general practitioner with prostate-specific antigen and digital rectal examination should be the primary goal. Currently, diagnosis is best established by transrectal ultrasound-guided biopsies. For the treatment of localized prostate cancer, men who undergo radical retropubic prostatectomy have been shown to have superior long-term results when compared to those who have received radiation therapy. With an improved understanding of the prostatic anatomy and nerve-sparing surgical techniques, morbidity from impotence and incontinence are minimal. In advanced carcinoma, 70 to 80% of men initially respond well to androgen withdrawal. Unfortunately, androgen-independent cells will continue to multiply, leading to tumor progression and death. Until effective chemotherapeutic agents are developed, we can only achieve palliation in advanced disease.


Flow cytometric analysis of reticulocytes using an RNA-binding fluorochrome. The reticulocyte count is an important parameter in the diagnosis of anaemia. The commonly used microscopic counting technique is, however, laborious and hampered by poor precision and low sensitivity. An alternative method has recently been developed, which is based on flow cytometric identification and quantification of reticulocytes after staining with an RNA-binding fluorochrome, thiazol orange. The two methods were compared both in a sample of healthy individuals and in patient specimens. The correlation between the methods was good, but the flow cytometric analysis consistently gave values approximately 1.5 times higher (reticulocytes in per cent of erythrocytes) than the microscopic technique. The coefficient of variation was significantly lower for the flow cytometric technique. Reference values for the reticulocyte count were determined and the effects of storage of specimens were evaluated. It is concluded that flow cytometric analysis of reticulocytes is more sensitive and has a better precision that the standard microscopic procedure.


Effect of prostaglandin synthesis inhibition on the tubuloglomerular feedback control in the rat kidney. It is known that intact prostaglandin synthesis is of some importance for tubuloglomerular feedback (TGF). In this study the effects of cyclooxygenase inhibition by oral fenflumizole, an imidazole derivative, on TGF in rats after acute elevation of ureteral pressure were investigated by comparison with the TGF mechanism in untreated rats. The effects of fenflumizole on the TGF mechanism were also compared with those of indomethacin injection. To characterize the TGF mechanism proximal tubular stop-flow pressure (Psf) was measured during perfusion of the loop of Henle with an artificial perfusion solution with a composition resembling that of the proximal fluid. The perfusion rate was varied from 0 to 40 nl.min-1, in steps of 2.5-5 nl.min-1, permitting measurement of the maximal pressure response delta Psf, to increased tubular perfusion, and the perfusion rate which elicited a half-maximal drop in Psf, designated the turning point (TP). Both cyclooxygenase inhibitors caused a significant increase in TP from a control value of 21 to approximately 27 nl.min-1. TP remained at this level in fenflumizole-treated rats, even after elevation of ureteral pressure, a situation in which TGF is normally not detectable. From these and earlier investigations we conclude that the prostaglandin system is of importance for a normally operating TGF control mechanism, both under normal conditions and, especially, in conditions in which resetting of TGF is observed.


Atrial natriuretic factor infusion following acute renal ischemia in anesthetized dogs. Atrial natriuretic factor (ANF) has been shown to be effective in reversing renal functional impairments following renal ischemia. We studied the effects of a nonhypotensive intravenous ANF infusion (100 ng/min x kgBW, 60 min) after 90 min unilateral renal arterial occlusion in anesthetized dogs with an intact contralateral kidney. ANF plasma levels remained unchanged in controls (group 1) and increased in ANF-infused animals (group 2) from 22 +/- 3 to 552 +/- 124 pg/ml. Blood pressure increased in both groups during renal ischemia, but returned to control values in group 2 when ANF infusion was started. Plasma vasopressin did not change in group 1, but increased in group 2 (0.77 +/- 0.29 vs. 1.10 +/- 0.49 pg/ml) after terminating ANF infusion. The postischemic fall in creatinine clearance (CCr), filtration fraction (FF) and renal blood flow (RBF) was prevented by infusion of ANF (CCr: group 1, 0.16 +/- 0.05 vs. group 2, 1.01 +/- 0.25 ml/min x kgBW; FF: group 1, 4.0 +/- 1.6 vs group 2, 14.1 +/- 4.1%; RBF: group 1, 6.0 +/- 1.2 vs. group 2, 9.2 +/- 1.6 ml/min x kgBW); however, the effects were limited to the time of infusion and the postischemic increase in urinary excretion of the proximal tubular enzyme N-acetyl-beta-D-glucosaminidase (NAG; group 1, 317.7 +/- 163.6 vs. group 2, 672.4 +/- 245.7 microU/min x kgBW) was not improved by ANF. Our data suggest that infusion of ANF transiently reverses postischemic renal impairment. However, the failure to demonstrate a sustained postischemic improvement of renal functional parameters and to ameliorate massive NAG excretion casts doubt on the benefit of ANF infusion in preventing cellular damage.


[Obsessive behavior and thoughts in patients with myasthenia gravis]. 230 patients with myasthenia gravis were investigated by a german questionnaire, the HAMBURGER-ZWANGS-INVENTAR, in order to quantify their obsessive and compulsive behaviour. Compared to normals, myasthenic patients show higher scores on the compulsion-subscales of the inventory. The most significant differences appeared on the orderliness-subscale. On the other hand ratings of obsessive behaviour are significantly lower than in the normal population. Especially (auto)aggressive ruminations do hardly occur in myasthenia, which is a remarkable fact in this autoaggressive immunological disease. Compulsive behaviour cannot be interpretated as clinical compulsive disorder, caused by myasthenia. Questionnaire replies rather indicate one aspect of the way of coping with the disease and its treatment.


Long-term renal handling of sodium and calcium in spontaneously hypertensive rats. The renal handling of sodium and calcium in spontaneously hypertensive rats (SHR) was investigated over an extended period (10-75 weeks of age) and compared with age-matched normotensive Wistar-Kyoto controls. The animals were fed a standard rat chow except during screening periods when liquid diet that matched the pellet chow was substituted. Sodium balance, urinary excretion of sodium and calcium, fractional excretion of sodium (FENa), and renal cortical dopamine receptors (DA1 and DA2) were measured at 10, 30, 60 and 75 weeks of age. The results showed no difference between the two strains except for FENa, which was significantly higher in the SHR at 75 weeks coincident with decreased glomerular filtration rate. We conclude that a defect in renal handling of sodium and/or calcium is not a major factor in the maintenance of hypertension in the SHR.


Permeability of the macula densa basement membrane area to high molecular weight molecules. To investigate the permeability properties of the basement membrane beneath macula densa cells and between extraglomerular mesangial cells, thick ascending limbs with attached glomeruli were dissected from rabbit kidney and incubated in a Ringers solution containing either horseradish peroxidase (HRP) (1 mg/ml; molecular weight approximately 40,000) or native or cationic ferritin (both at 5 mg/ml molecular weight approximately 450,000) for 5 and 20 min at room temperature. Tubules were processed for electron microscopy. At both time points, HRP reaction product fully permeated the matrix material between extraglomerular mesangial cells, the basement membrane underneath the macula densa, and also was occasionally located in intercellular spaces and intracellular vesicles within macula densa cells. Similar results were obtained with native and cationic ferritin. In separate experiments, thick ascending limbs with attached glomeruli were perfused for 20 min at room temperature with a Ringer solution containing HRP (1 mg/ml). HRP was found in tubulovesicular bodies within the apical cytoplasm of macula densa cells again indicating that these cells exhibit endocytotic activity. However, HRP did not gain access to intercellular spaces indicating that the apical junctional complex was impermeable to HRP. These results demonstrate that the macula densa basement membrane and matrix material between extraglomerular mesangial cells is permeable to high molecular weight molecules and suggest unhindered diffusion of water and solutes within this area.


[Etiology, follow-up and therapy of seizure clusters in temporal lobe epilepsy and catamenial epileptic seizures]. The phenomenon of seizure clustering is still poorly understood. We therefore investigated 192 patients with temporal lobe epilepsies among whom 60 showed clustering of seizures. The percentage of women was significantly higher in the cluster than in the non-cluster group, the history of epilepsy lasted longer and the excess of complex partial seizures over tonic clonic seizures was more prominent in the cluster group. In 46 out of the 60 patients the clustering did not occur initially but developed in the course of the disease. In a particular subgroup the development initiated with isolated tonic clonic seizures, in a later phase complex partial seizures appeared and finally only complex partial seizures remained. This type of history was found significantly more frequent in the cluster than in the non-cluster group (27% versus 7%). It is conjectured that endogenous, as well as exogenous factors, both of them not completely revealed, cause the occurrence of clusters; anticonvulsant drug therapy might even enlarge this trend. Patients with seizure clustering tend to be pharmacoresistant. Chronic therapy with antiepileptic drugs besides intermittent therapy with benzodiazepines may help. A particular type of seizure clustering is observed in catamenial epilepsies where seizures appear in the perimenstrual and/or periovulatory phase of the menstrual hormonal cycle of females. This type of seizure incidence is obviously influenced by hormonal rhythms. Ten patients suffering from catamenial epileptic seizures were therefore treated with a synthetic analogue of GnRH in order to suppress the menstrual hormonal rhythm. As a result 3 patients became seizure free and in 5 patients seizure frequency decreased.


Mechanism of acetazolamide-induced rise in renal vascular resistance assessed in the dog whole kidney. The reduction in renal blood flow (RBF) and glomerular filtration rate (GFR) observed after the administration of the carbonic anhydrase inhibitors acetazolamide and benzolamide had been explained as due to activation of the tubuloglomerular feedback mechanism. If correct, pharmacologic blockade of this pathway should prevent the development of renal vasoconstriction with the carbonic anhydrase inhibitors. Thus, the current study evaluates in the dog whole kidney the effect of acetazolamide (20 mg/kg body weight) in the presence or absence of furosemide (5 mg/kg body weight), a drug which blocks the tubuloglomerular feedback. Acetazolamide resulted in a large increase in urinary bicarbonate excretion accompanied by a significant reduction in GFR (16%) and RBF (18%). By contrast with the effects of acetazolamide, furosemide did not alter GFR and increased RBF. In addition, the loop diuretic induced a large chloruresis without changes in urinary bicarbonate excretion. The infusion of acetazolamide in furosemide-treated dogs resulted in a significant increment in renal bicarbonate excretion and in a significant reduction in the levels of both GFR (28%) and RBF (13%). Therefore, furosemide pretreatment did not block the effects of acetazolamide on renal hemodynamic parameters. Consequently, the acetazolamide-induced reduction in both GFR and RBF cannot be accounted for by changes in chloride levels in the juxtaglomerular region due to enhanced salt transport in the macula densa/distal nephron. The increased renal vascular resistance observed with acetazolamide might occur by either a direct effect of this agent on the renal circulation or as a result of changes in intrarenal pressure secondary to the inhibition of proximal fluid reabsorption.


[Graduate education for psychiatry and psychotherapy specialty FMH: retrospective evaluation by the psychiatrist. 1]. Psychiatrists who recently finished their postgraduate training filled in a questionnaire about their training experiences. Two thirds considered the training as generally insufficient although many gave high ratings when asked about the quality of their clinical experiences and the acquisition of skills. It appears that the main areas for possible improvement of the training should concern knowledge and practical experiences with disorders which are typical for psychiatric out-patients.


[Graduate education in psychiatry and psychotherapy specialty FMH: retrospective evaluation by psychiatrists. 2]. A questionnaire filled in by psychiatrists shortly after the end of their postgraduate training showed that the majority invested a considerable amount of time and money for a psychotherapy training on private initiative. Three quarters had a training analysis. About 70% felt that their private training efforts had been more important than the official psychotherapy training in the institutions. Psychiatrists working in institutions tended towards a more positive rating of the official postgraduate training.


[Environmental dependence in brain lesions: imitative behavior, prehension and utilization]. This paper reviews the literature on clinical signs such as imitation behavior, grasp reaction, manipulation of tools, utilization behavior, environmental dependency, hyperlexia, hypergraphia and echolalia. Some aspects of this semiology are of special interest because they refer to essential notions such as free-will and autonomy.


[Comparison of urography and ultrasonography findings in patients prior to prostatectomy]. In order to evaluate the assess of ultrasonographic examinations in patients before prostatectomy the authors correlated preoperative findings obtained by urography with ultrasonographic findings. Urography of the upper urinary pathways revealed 12 (18.8%) pathological findings, ultrasonographic examination 26 findings (40.6%) in a group of 64 patients. While in 14 patients a pathological finding on ultrasonography was not revealed by urography, only in one patient with a positive urographic finding ultrasonography was negative. Ultrasonographic examination proved to be more valuable in assessing the post-miction residue, as compared with urography. Moreover, ultrasonography is useful for assessing the volume of tissue of benign prostatic hyperplasia. Correlation of the calculated volume of prostatic tissue by means of ultrasonography with the weight of removed tissue revealed a close linear correlation. Ultrasonographic examination of patients before prostatectomy replaced urography which proved to be unnecessary diagnostic procedure.


Transhiatal bilateral splanchnicotomy for pain control in pancreatic cancer: basic anatomy, surgical technique, and immediate results in fifty-one cases. The greater splanchnic nerves are largely responsible for innervation of the supramesenteric viscera; their section is known to be efficient to relieve pancreatic pain. Transhiatal splanchnicotomy (THS) is easily performed through a midline laparotomy. The nerve trunks are readily identified in the submediastinal space, far from the pancreatic cancer motivating splanchnicotomy, and can be sectioned safely and completely. After carrying out an anatomic study to determine the level of origin and mode of constitution of the greater splanchnic nerve trunk and its relations to the posterior and lower mediastinum, 51 patients underwent THS for intractable pain caused by unresectable pancreatic adenocarcinoma. THS alone was performed in 22 cases. THS was performed in association with biliary tract diversion or gastroenteroanastomosis in the other cases. All tumors were considered unresectable during surgery, and no patient was operated on with the sole purpose of performing THS. Two deaths (3.9%) were unrelated to THS. Specific morbidity was 6% (one pneumothorax, one chylothorax, and one splenic injury). Immediate postoperative functional results were good in 86.3% of patients treated by THS alone (group 1) and in 80.7% of patients treated by THS and bypass (group II). Functional results decreased to 72.7% in group I and 62.1% in group II, 3 months after surgery. In conclusion, THS appears to be an efficient technique for relief of pancreatic neoplastic pain and need not be combined or confused with medical percutaneous methods of neurolysis.


Repair of posterolateral ventral herniation caused by diabetic truncal neuropathy. Abdominal pain caused by diabetic radiculopathy is uncommon, and abdominal hernia as a complication is rare. We report a case of abdominal wall herniation caused by diabetic truncal radiculopathy requiring surgical repair. Clinical and diagnostic features of this entity are reviewed.


Biological properties of a crude venom extract from the greater weever fish Trachinus draco. Crude venom of the greater weever fish, Trachinus draco was analyzed to assess its toxicity, stability and biological properties. The best yield of venom was obtained by extraction in physiological saline of the whole venom apparatus of the fish which were shock-frozen and stored at -70 degrees C. This extract had a mouse i.v. minimum lethal dose of 1.8 micrograms protein per gram mouse and a total of 61,000 minimum lethal doses were obtained from venom apparatus of one fish. The lethal activity was unstable at room temperature especially at lower protein concentrations. Stability was achieved either by storing the extract at -70 degrees C or by precipitation with ammonium sulfate at 50% saturation. Toxicity of the crude venom was abolished by trypsin treatment. The crude venom did not possess any proteolytic or histamine-releasing activities. The venom caused an outflow of tetraphenylphosphonium from preloaded rat brain particles in a concentration-dependent manner. Like toxicity, this effect was also abolished by trypsin treatment or by keeping the venom at higher temperatures. The crude venom also possessed hemolytic activity with an EC50 for rabbit erythrocytes of 75 ng/ml venom protein. The hemolytic activity was also sensitive to heat and proteolytic treatment. Rabbit erythrocytes were most sensitive to venom followed by rat erythrocytes. Mouse and cattle erythrocytes were only slightly sensitive, whereas human, chicken and guinea pig erythrocytes were totally resistant.


Temporal relation of calcium-calmodulin binding and neuronal damage after global ischemia in rats. This study explores the temporal relation of the severity of ischemia and calcium-calmodulin binding in vulnerable and resistant brain regions in a commonly used model of global ischemia. Immunohistochemical assay of free calmodulin unbound to calcium and light microscopic histological damage were measured in rats after 5, 10, or 20 minutes of global ischemia. After 24 hours of reperfusion, decreased calmodulin staining, representing increased calcium influx and calcium-calmodulin binding, correlated with increasing durations of ischemia across all brain regions. Based on a 4-point scale (4, extensive stain; 0, no staining), calmodulin staining after 5 minutes versus 10 minutes of ischemia was 3.2 versus 1.9, respectively (p less than 0.05) and after 10 minutes versus 20 minutes of ischemia was 1.9 versus 1.0, respectively (p less than 0.01). The CA1 region displayed the greatest sensitivity to ischemia. Similar but less dramatic results were seen after 2 hours of reperfusion. After 72 hours of reperfusion, histological damage closely correlated with calcium-calmodulin binding after variable durations of ischemia. A threshold of 10 minutes of ischemia was required to cause calcium-calmodulin binding and irreversible neuronal damage. Surviving neuronal populations showed recovery of calmodulin staining 7 days after ischemia, representing a return of free calmodulin and normal calcium homeostasis. These correlations between calcium-calmodulin binding, histological damage, and duration of ischemia support the causal role of calcium influx in global ischemic injury and suggest the need for very rapid intervention after ischemia if calcium-mediated damage is to be prevented.


Intrinsic lethality of chloride-channel-directed insecticides and convulsants in mammals. The intrinsic lethality of a series of chloride-channel-directed convulsants and insecticides was determined by intracerebral injection into mice. The toxicities of the cyclodiene insecticides and picrotoxinin were potentiated by intracerebral injection, when compared to their reported intraperitoneal LD50s. In contrast, the toxicities of lindane, abamectin, and the bicyclic convulsants TBPS and a TBOB analog were approximately the same by either route of administration. These results suggest that the brain is the primary target site for the cyclodienes and picrotoxinin, while the peripheral nervous system may be relatively more important in the toxic action of lindane, abamectin, and bicyclic convulsants such as TBPS. With the exception of picrotoxinin these compounds showed, overall, a good correlation between acute intracerebral toxicity and potency for inhibiting GABA-dependent chloride uptake. The evidence that multiple chloride-channel subtypes serve as targets for these compounds and the potential impact this had on the results of these studies are discussed.


The varieties of ribonuclease P.

The varieties of ribonuclease P. Ribonuclease P is a ribozyme involved in tRNA processing that is present in all cells and organelles that synthesize tRNA. Most of our understanding of ribonuclease P derives from studies of the bacterial enzyme. This enzyme has been characterized biochemically and a secondary structure for the RNA subunit has been proposed. Isolation and characterization of ribonuclease P from diverse Archaea and Eukarya are now modifying and adding to our model of this unusual enzyme. The latter instances of RNase P differ from the bacterial version, but similarities are emerging.


Antigenic relationships of foot-and-mouth disease virus serotype Asia-1 isolates demonstrated by monoclonal antibodies. A panel (26) of monoclonal antibodies (MAbs) was elicited against three distinct isolates of foot-and-mouth disease virus (FMDV) serotype Asia-1. Each MAb was characterized according to the location of its epitope: Class I, restricted to the intact virion (140S); Class II, restricted to 140S and the virion protein subunit (12Sps); Class III, available on 140S, 12Sps and virus protein 1; Class IV, restricted to 12Sps. In addition, the MAbs were further categorized by isotype, neutralization of viral infectivity, capacity to bind in radioimmunoassay and precipitation in the Ouchterlony reaction. Neutralization of FMDV infectivity by a MAb of the IgA isotype is reported for the first time. A minimum of seven distinct neutralization epitopes were described on FMDV Asia-1. Some of the neutralizing MAbs bound FMDVs in addition to those that they neutralized. The MAbs defined epitopes common to FMDV serotypes Asia-1, A, O1 and C but neutralizing capacity was restricted to serotype Asia-1. Class IV MAbs defined epitopes highly conserved throughout the FMDV serotypes. Identification of FMDV neutralization epitopes makes possible the direct selection of optimal FMDV strains for vaccine fabrication. In addition, these data are crucial to the design of future synthetic vaccines.


Proliferating cell nuclear antigen in malignant and pre-malignant lesions of epithelial origin in the oral cavity and the skin: an immunohistochemical study. Proliferating cell nuclear antigen (PCNA) is a nuclear protein synthesized in the late G1 and S phase of the cell cycle and immunohistochemical detection of the protein represents a useful marker for the proliferating fraction of cells in tissue specimens. A series of malignant and pre-malignant lesions of the oral cavity and skin were evaluated by the streptavidin biotin immunoperoxidase method for detection of this protein. Monoclonal anti-PCNA antibody (PC 10) labelled proliferating cells in all cases with varying intensity of nuclear staining. In squamous cell carcinoma (n = 48), PCNA positivity correlated with the differentiation and atypia of the tumour cells; however, in poorly differentiated tumours, the relationship between PCNA expression and proliferation was lost. Basal cell carcinoma showed an increased growth fraction in tiny epithelial nests (mean 43.8, SD 6.0, n = 20) than in neoplastic basal cells (mean 30.1, SD 6.9, n = 8). The growth fractions were significantly higher in the pre-malignant lesions (leukoplakia, mean 22.3, SD 7.7, n = 14; Bowen's disease, mean 45.2, SD 11.7, n = 12; senile keratosis, mean 41.2, SD 7.0, n = 12) than in the normal mucosa (mean 9.8, SD 4.9, n = 10), suggesting that cellular growth fractions correlate with the degree of dysplasia in pre-malignant lesions.


Unusual prevalence of the rare serovar Da of Chlamydia trachomatis in Greece detected by monoclonal antibodies. Twenty-seven monoclonal antibodies (mAb), eight with subclass-specificities and nineteen reacting with one or two C. trachomatis serovars, were developed and used to immunotype twenty-six clinical isolates from Greek patients with chlamydial infections. Twelve samples, first classified as serovar D, were identified as the recently established serovar Da on the basis of their negative reaction with mAb JG9, an mAb previously shown to distinguish between D and Da, and a new mAb 114D9 with similar specificity. The 12 Da strains represent the highest prevalence of the rare serovar Da that has been reported worldwide. Further characterization of mAbs JG9 and 114D9 revealed a unique cross-reactivity of the D-specific JG9 with the mouse biovar MoPn. Epitope mapping studies on overlapping synthetic peptides of the fourth variable domain of the MOMP of serovar D and Da localized the epitopes of JG9 and 114D9 and a D/Da specific mAb 113D5 within this domain. The results were consistent with the specificity of these mAbs observed with whole microorganisms and indicated their usefulness as epidemiologic markers.


Expression of c-erbB-2 protein and epidermal growth factor receptor in normal tissues of the female genital tract and in the placenta. The c-erbB-2 (HER-2/neu) protein is a membrane glycoprotein growth factor receptor that has molecular homology with the epidermal growth factor receptor (EGFR). To investigate the relationship between the expression of c-erbB-2 protein and EGFR in the tissues of the human female genital tract and in the placenta, we examined the immunohistochemical reactivity of monoclonal antibodies against both of these proteins. In the müllerian-derived genital tract, epithelial cells of the fallopian tube, endometrium, and endocervix showed reactivity for c-erbB-2 protein, whereas reactivity for EGFR was distributed mainly in the stromal cells throughout the menstrual cycle and during pregnancy. In addition, the staining intensity for EGFR in the endometrial stroma increased with its decidualization. In the exocervical squamous epithelium, basal cells were c-erbB-2 protein-negative and EGFR-positive, but the more differentiated squamous cells of the intermediate layer were c-erbB-2 protein-positive and EGFR-negative. In the placental tissues, cytotrophoblasts and syncytiotrophoblasts of the chorionic villi were c-erbB-2 protein-negative and EGFR-positive. In contrast, intermediate trophoblasts in the extravillous space were c-erbB-2 protein-positive and EGFR-negative. Thus, there is an inverse relationship between the expression of c-erbB-2 protein and EGFR in the tissues of the female genital tract and in the placenta. This suggests that there may be a regulatory mechanism(s) for the expression of both proteins that is associated with the differentiation and/or function of cells in the female genital tract and the placenta.


Light and electron microscopic immunolocalization of endothelial leucocyte adhesion molecule-1 in inflammatory bowel disease. Morphological evidence of active synthesis and secretion into vascular lumen. Endothelial leucocyte adhesion molecule-1 (ELAM-1) is a rapidly inducible adhesion molecule for neutrophils in vascular endothelial cells. We investigated its immunohistochemical localization in 17 cases of inflammatory bowel disease. ELAM-1 was preferentially expressed in venules in actively inflamed mucosa and granulation tissue. Most capillaries were negative for ELAM-1. In areas with mild inflammation its expression diminished markedly and in normal mucosa of the colon and small intestine its expression was sparse. Electron microscopically, venules in active inflammation had swollen endothelial cells with well-developed rough endoplasmic reticulum. Immunoelectron microscopy revealed ELAM-1 localization along the luminal plasma membrane and in rough endoplasmic reticulum and round granules, findings suggestive of active production in endothelial cells. Furthermore, exocytosis of immunoreactive substance into the lumen was confirmed. Our study suggests that venules in actively inflamed area play an important role in eliciting and/or maintaining acute inflammatory processes by active permeation of neutrophils from the blood stream into the tissue, and that ELAM-1 may be a secretory protein as well as a transmembrane receptor protein.


The distribution and possible function of gamma interferon-immunoreactive cells in normal endometrium and myometrium. T-lymphocytes are present in normal endometrium, where they may have a role in the control of glandular maturation. T-cell activity could be related to the local secretion of cytokines such as gamma interferon, which has an anti-proliferative effect on endometrial epithelial cells in vitro. We have examined gamma interferon immunoreactivity and T-cell distribution in 24 normal pre-menopausal uteri. Endometrial appearances were representative of all stages of the menstrual cycle. Most cells in the lymphoid aggregates in the stratum basalis were stained by T-cell and gamma interferon antisera. T-lymphocytes were also scattered in glandular epithelium and throughout the stroma of basal and functional layers; immunoreactivity for gamma interferon was less consistent in these cells. There was no alteration in the intensity or distribution of gamma interferon staining in different phases of the menstrual cycle. Endometrial granulocytes (K-cells) present mainly in the late secretory endometria were not reactive with the gamma interferon antiserum. In addition to endometrial staining, T-cells were distributed in all areas of the myometrium in most uteri, and many myometrial lymphocytes were gamma interferon positive. These results support a role for gamma interferon in endometrial physiology, possibly as an inhibitor of epithelial proliferation.


Basement membrane proteins in salivary gland tumours. Distribution of type IV collagen and laminin. Immunohistochemical localization of type IV collagen and laminin in normal salivary glands and in salivary gland tumours of various types was studied using rabbit antisera. In normal salivary glands, type IV collagen and laminin were co-localized in basement membranes surrounding acini, ducts, fat cells and peripheral nerves. In salivary gland tumours, three main patterns of co-expression of these basement membrane proteins were distinguished. Linear basement membrane-like staining was detected in duct-cell-derived benign salivary gland tumours and in acinic cell carcinomas. In invasive lesions, however, these basement membrane proteins were distributed in an irregular, interrupted manner, and in many cases they were completely absent. Both benign and malignant salivary gland tumours which have a prominent myoepithelial cell component display a particular deposition of basement membrane molecules adjacent to the modified myoepithelial cells, and at the margins of extracellular matrix deposits within these tumours.


Nucleolar organizer regions of megakaryocytes in chronic myeloproliferative disorders. To study megakaryocyte activation, the argyrophilic staining method of nucleolar organizer regions (AgNOR) has been applied to decalcified bone marrow biopsies of 16 individuals with no haematopoietic disorders and 59 patients with chronic myeloproliferative disease. Of the 59 patients, 18 had chronic myeloid leukaemia (CML), 21 chronic megakaryocytic granulocytic myelosis (CMGM), 13 polycythaemia vera (PV) and 7 essential thrombocythaemia (ET). The AgNOR number of megakaryocytes in CML was significantly lower, and in CMGM, PV and ET significantly higher than in healthy individuals. The high number and the clusters of fine-grained AgNORs of megakaryocytes in CMGM, PV and ET are suggestive of active, proliferating cells. The AgNOR number of megakaryocytes and the platelet counts of the patients did not show a convincing correlation. In CMGM, PV and ET the pyknotic, heterochromatinized megakaryocytes with narrow rims of cytoplasm called bare (nude) nuclei, possessed few, large AgNOR granules. The AgNOR staining of bare nuclei and the roughly identical number of granules found in CMGM, PV and ET indicate a common, active mechanism of apoptosis.


Progesterone and promegestone stimulate human bone cell proliferation and insulin-like growth factor-2 production. Recent clinical studies suggest that progesterone may be involved in the regulation of bone turnover and could promote bone formation. This study was undertaken to evaluate whether progesterone and promegestone (a 19 nor-PG derivative) may have a direct effect on human bone cells and, if so, whether growth factor production could be involved in promoting this effect. The osteosarcoma cell line TE85 and untransformed normal human osteoblastic cells derived from iliac crest were used as in vitro model systems. Progesterone and promegestone were found to significantly increase [3H]thymidine incorporation in TE85 cells in a dose-dependent manner at concentrations ranging from 10(-12) to 10(-8) mol/l after four days of cultivation (p less than 0.01, ANOVA). Consistent with this response in the TE85 cells, progesterone and promegestone increased cell number in human osteoblastic cells after six days of treatment (p less than 0.05, ANOVA). To determine whether this effect on cell proliferation was mediated by the insulin-like growth factor (IGF) regulatory system, the levels of IGF-1, IGF-2 and IGF binding protein (IGFBP) were measured in the conditioned media of both TE85 and human osteoblast cells. While no significant changes in IGF-1 levels were found in the conditioned media of progesterone and promegestone treated cultures, progesterone and promegestone at the concentration of 5 nmol/l significantly increased IGF-2 levels 2.4 and 1.5-fold respectively, at 48 h in the conditioned medium of TE85 cells as compared to control. Similarly, a 4.1 and 1.9-fold increase in IGF-2 levels was found upon treatment with progesterone and promegestone in human osteoblastic cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Risperidone versus haloperidol in the treatment of chronic schizophrenic inpatients: a multicentre double-blind comparative study. Forty-four chronic schizophrenic inpatients participated in this multicentre 12-week parallel-group double-blind trial. After a run-in period of 2 weeks and a single-blind placebo wash-out of 1 week, they were randomly assigned to treatment with either the serotonin2 and dopamine-D2 antagonist risperidone or haloperidol. Two patients were excluded from the efficacy analysis. Five patients dropped out in the haloperidol group and 1 in the risperidone group. At the end of the trial, the mean daily dose was 12 mg for risperidone and 10 mg for haloperidol. The risperidone group showed greater improvement on the Positive and Negative Syndrome Scale for Schizophrenia, the Schedule for Affective Disorders and Schizophrenia-change version, and the Nurses' Observation Scale for Inpatient Evaluation. The improvement of negative symptoms was more pronounced in the risperidone group until week 8 of double-blind treatment. The consumption of antiparkinsonian medication was 10 times lower with risperidone. Both drugs were well tolerated and the laboratory, endocrinological and cardiovascular safety parameters were comparable. This study suggests that risperidone is comparable to haloperidol as an antipsychotic, but that it has a safer EPS profile.


Ventricular enlargement, clinical correlates and treatment outcome in chronic schizophrenic inpatients. The ventricle-brain ratio (VBR) of 42 chronic schizophrenic patients was compared with that of 42 age-matched medical controls. For the schizophrenics, the relationship of various clinical parameters to the VBR was assessed, and the outcome of 12 weeks of double-blind treatment with either risperidone or haloperidol. The results confirm that schizophrenic patients have slightly enlarged lateral ventricles compared with medical controls. Only for schizophrenics, an effect of age, but not of duration of illness, was noticed. This study does not support the validity of a clinical subdivision of chronic schizophrenic patients on the basis of the VBR. Neither negative, positive nor general psychopathological symptoms, as measured by the Positive and Negative Syndrome Scale for Schizophrenia (PANSS), were related to the VBR, nor were abnormal involuntary movements or extrapyramidal symptoms. No association between season of birth or a family history of major mental disorder and VBR could be demonstrated. Treatment response was predicted by the total PANSS score and the PANSS general psychopathology subscale score at baseline. There was a trend for patients with higher VBR to have a more or haloperidol). or haloperidol).