[Liver transplantation for hepatocellular carcinoma. Mid-term results]. The results of liver transplantation for hepatocellular carcinoma are disappointing. Thus, in 11 patients transplanted for hepatocellular carcinoma on cirrhosis, the actuarial survival rate at 3 years was 20 percent, with a postoperative mortality of 27 percent and 4 deaths caused by recurrence of the malignancy. These poor results contrast with those we obtained after liver transplantation for cirrhosis and finding of incidental hepatocellular carcinoma in the resected part of the liver. Out of 6 patients with incidental carcinoma only 1 died postoperatively (16.6 percent), and no case of recurrent malignancy was observed. These results confirm that the malignancy recurrence rate is high after transplantation for large carcinoma. On the other hand, the absence of recurrence after transplantation for incidental hepatocellular carcinoma encourages us to look for small carcinomas on cirrhosis.


Characterization of amylin binding sites in a human hepatoblastoma cell line. Amylin binding sites in a human hepatoblastoma cell line (HepG2) have been characterized in detail. 125I-Amylin (rat) bound to HepG2 cells with high affinity. Binding was reversible and selective, and dependent on time and temperature. Scatchard analysis revealed the presence of high (Kd = 0.11 +/- 0.04 nM) and low (Kd = 1.3 +/- 0.4 microM) affinity binding sites for 125I-amylin in HepG2 cells. The dissociation experiments also showed that 125I-amylin dissociated from high- and low-affinity sites. The association data, however, indicated the presence of only one binding site. Rat amylin was more potent than human amylin and rat calcitonin gene-related peptide (CGRP) in displacing 125I-amylin bound to HepG2 cells. Nonhomologous peptides did not displace 125I-amylin. Rat amylin was, however, less potent than rat CGRP in displacing 125I[Tyr0]CGRP from HepG2 cells. Pretreatment of HepG2 cells with rat amylin (10 nM) reduced the specific binding of 125I-amylin by 75%, whereas rat CGRP (10 nM) pretreatment had no effect on amylin binding. Calcitonin gene-related peptide, as well as rat and human amylin, stimulated the adenylate cyclase activity of HepG2 cell membrane preparation in a dose-dependent manner, with an order of potency of CGRP > rat amylin > human amylin. A CGRP antagonist, CGRP(8-37), significantly attenuated the stimulatory effect of both amylin and CGRP on adenylate cyclase activity. These investigations show that distinct receptors of amylin and CGRP are present in HepG2 cells, and that amylin stimulates adenylate cyclase activity through CGRP receptors. This system could now be exploited for studying amylin receptors and amylin-mediated signal transduction.


Characterization of the novel CCK analogs JMV-180, JMV-320, and JMV-332 in H345 cells. The interaction of the novel CCK analogs JMV-180, JMV-320, and JMV-332 with CCK-B/gastrin receptors on small cell lung cancer (SCLC) cells was investigated. JMV-180, JMV-320, and JMV-332 potently inhibited specific binding of 125I-CCK-8 to CCK-B/gastrin receptors expressed on the SCLC cell line NCI-H345 (H345) with IC50 values of 4.9, 1.8, and 7.0 nM, respectively. JMV-320 and JMV-332 stimulated intracellular calcium ([Ca2+]i) release in a dose-dependent manner in cells preloaded with indo-1. JMV-180 did not stimulate [Ca2+]i but inhibited the [Ca2+]i release elicited by 10 nM CCK-8 in a dose-dependent manner. These data indicate that JMV-320 and JMV-332 function as CCK-B/gastrin receptor agonists while JMV-180 functions as a CCK-B/gastrin receptor antagonist in H345 cells.


[Drug addiction and hepatitis C: does a correlation exist between the laboratory and clinical pictures?]. The prevalence of anti-HCV antibodies reaches 90% positive among intravenous drug addicts. The authors evaluate the possibility of relating laboratory findings to clinical evolution of hepatitis C. Besides, a comparison between first generation anti-HCV system ELISA and the newer RIBA detection system showed no significant difference.


Interaction of opioid peptides and other drugs with multiple delta ncx binding sites in rat brain: further evidence for heterogeneity. Recent pharmacological data strongly support the hypothesis of delta receptor subtypes as mediators of both supraspinal and spinal antinociception (delta 1 and delta 2 receptors). In vitro ligand binding data, which are fully supportive of the in vivo data, are still lacking. A previous study indicated that [3H][D-Ala2,D-Leu5]enkephalin labels two binding sites in membranes depleted of mu binding sites by pretreatment with the site-directed acylating agent, 2-(p-ethoxybenzyl)-1-diethylaminoethyl-5-isothiocyanatobenzimid azole-HCI (BIT). The main goal of the present study was to develop a ligand-selectivity profile of the two delta ncx binding sites. The data indicated that naltrindole and oxymorphindole were relatively selective for site 1 (20-fold). [D-Ser2,Thr6]Enkephalin and deltorphin-II were only 2.7-fold and 2.2-fold selective for site 1. [D-Pen2,D-Pen5]Enkephalin and deltorphin-I were 80-fold and 38-fold selective for site 2. 3-Iodo-Tyr-D-Ala-Gly-Phe-D-Leu was 52-fold selective for site 1. Morphine had moderate affinity for site 1 (Ki = 16 nM), and was about 11-fold selective for site 1. Thus, of the 10 drugs studied, only DPDPE and DELT-I were selective for site 2. Viewed collectively with other data, it is likely that the delta 1 receptor and the delta ncx binding site are synonymous.


[Pharmacological treatment of neuropathic pain]. Neuropathic pain, i.e., pain resulting from functional changes in peripheral and central pathways subsequent to injury to the peripheral nervous system, offers a most difficult challenge to therapy. To date, only the antidepressants and the anticonvulsants have shown any effectiveness, albeit incomplete and inconsistent, and many questions remain unanswered: What are the exact indications for the antidepressants? What component of neuropathic pain do they relieve, and through which mechanisms? Which type of antidepressants should be prescribed? A first-generation tricyclic? Or a new compound with a selective action on serotonin reuptake? What are the effective dosage and duration of the treatment? What is it mechanism of action? What other antalgic effects do carbamazepine and baclofen possess apart from their action on trigeminal neuralgia? The opiates are generally considered to be without effect, but recent clinical and experimental findings seem to point otherwise. In the meantime, following a few simple rules will optimize the benefit of drug treatment in neuropathic pain: treatment tailored to individual cases; adequate dosage and duration of treatment. However, it is from the near future that breakthroughs are being expected, dues to the multiplication of animal models and more accurate analysis; new clinical evaluation tools which help in distinguishing the different mechanisms underlying the various aspects of pain; the development of new substances, such as capsaicin, local anesthetics, anti-inflammatory agents (NSAIDs for example); and better defined methodological conditions for therapeutic trials.


[Cyclic nucleotides in rat tissues under long-term irradiation]. Considerable changes in concentrations of cAMP and cGMP and in the CAMPOcGMP ratio were found in the thymus and blood plasma of rats subjected to long-term irradiation at cumulative doses of 10 to 20 Gy. This might be a manifestation of adaptation reactions that develop in response to the effect of ionizing radiation.


Unfavorable experience with hypofractionated radiotherapy in unresectable lung cancer. The use of a reduced number of large-sized fractions in radiotherapy (hypofractionation) is usually associated with poor therapeutic results and severe adverse effects, in accord with radiobiologic concepts. However by some authors unresectable lung cancer patients have been treated with hypofractionated radiotherapy with the main aim of "convenience". Result and damage rates are reported to be comparable to those of conventional treatment. In our experience, based on palliative irradiation of 86 advanced-stage, nonmicrocytoma patients, objective remission rates, subjective and performance status improvement, and survival overall were as poor as could be expected in this kind of presentation, with no striking impact of this treatment modality. Severe adverse effects were shown by a large proportion of cases involving skin and soft tissues of the chest wall (40%) and lungs (55.5%). The incidence of severe damage was in agreement with BED (biologic effective dose) values, differently from other experiences of radiotherapeutic management of advanced lung cancer with large fractions.


Cyclophosphamide, epirubicin, etoposide, cis platinum (CEVP) in combination with radiotherapy: evaluation of a protocol adopted for 6 years in 148 cases of small cell lung cancer. The present study was carried out with the purpose of assessing and evaluating the responsiveness of small cell lung cancer (SCLC) to combined treatment (i.e., chemotherapy and radiotherapy). This approach was applied to all patients observed in our Institution during the last 6 years with the exclusion of patients more than 65 years old, with Karnofsky performance status less than 60, and with concomitant non-neoplastic diseases. One hundred and forty-eight patients were selected and treated with a combination chemotherapy regimen including cyclophosphamide, etoposide, epirubicin and cisplatinum. For patients with limited disease, treatment consisted of 3 cycles of chemotherapy (induction), followed by radiation therapy and then by another 3 cycles of chemotherapy (consolidation). For those with extensive disease, treatment consisted only of 6 consecutive cycles of chemotherapy. Results, besides showing the good activity and tolerability of the chemotherapeutic regimen employed (which induced 80% overall response), indicated that some prognostic factors such as the number of metastatic sites, brain and liver involvement and performance status are essential in determining the outcome of response and, particularly, of survival. In addition, the efficacy and tolerability of the combination chemotherapy-radiotherapy were confirmed, as was the evidence of a synergistic pharmacologic effect between cis-platinum and etoposide.


[Significance of ED50 and therapeutic indexes of various radiation-protective agent groups]. Radioprotective agents are divided in 3 groups: (1) cystamine, AET, cystaphos, gammaphos, and thiogammaphos with ED50 (the dose that gives a half of the maximal protective effect) of 10(3)-10(1.6) mumol/kg and therapeutic index K = LD50/ED50 = 10(0)-10(1.6); (2) 5-methoxytryptamine, phenylephrine, serotonin, and norepinephrine with ED50 = 10(1)-10(0) mumol/kg and K = 10(1.8)-10(2,6); (3) clonidine and isoprenaline with ED50 = 10(-0.5)-10(-0.8) mumol/kg and K = 10(3)-10(4). Possible causes of these differences and advantages of low ED50 and high K are discussed.


Opportunistic intraocular infections in AIDS. In conclusion, this clinicopathologic study has shown that CMV ocular infection is present in about 16% of terminal AIDS patients. The treatment of CMV retinitis reduces the number of CMV-infected nonocular organs and may also lessen the severity and control the spread of concurrent nonocular infection, both of which may prolong survival in AIDS patients. Other opportunistic infections, involving primarily the choroid, were also seen in a number of patients, some of whom had concurrent intraocular infections with CMV and P carinii, M avium-intracellulare, C neoformans. In addition, all of these choroidal infections were components of disseminated infection, underscoring the increasingly important role of the ophthalmologist in the diagnosis and treatment of disseminated opportunistic infections in AIDS.


A recombinant-based feline immunodeficiency virus antibody enzyme-linked immunosorbent assay. We have developed an antibody detection enzyme-linked immunosorbent assay (ELISA) for the identification of animals infected by feline immunodeficiency virus (FIV). The ELISA solid-phase antigen consists of recombinant FIV gag proteins expressed in bacteria. The proteins are purified from bacterial lysates as insoluble inclusion bodies. In the case of bacterially expressed p24gag, it is shown that all of the linear, sequential epitopes presented by viral p24 during infection are retained. Purified preparations can be substituted for solid-phase whole virus in the IDEXX PetChektm immunoassay. The antibody ELISA duplicates the sensitivity and specificity of the whole virus based PetChek plate assay.


Long-term clinical observations on feline immunodeficiency virus infected asymptomatic carriers. Eleven feline immunodeficiency virus (FIV) infected asymptomatic carrier (AC) cats were observed for 2 years for development of acquired immunodeficiency syndrome (AIDS). Four of the 11 (36.4%) showed progression of the clinical stage. Persistent generalized lymphadenopathy was noted in three cats as the first sign of illness after the AC phase, while the other showed lymphadenopathy with signs of AIDS-related complex. In all four cats the AIDS-related complex stage lasted for 10 months or longer, and two showed progression of the disease into AIDS. The two cats showing AIDS illnesses died within approximately 1 year after they had developed persistent generalized lymphadenopathy. Pathology confirmed the diagnosis of AIDS characterized by the presence of depletion lesions in the lymphoid organs, and of severe infections of an opportunistic nature. The overall mortality of FIV infected AC cats during a 2 year period was two out of 11 (18.2%). These cats showed decreased concanavalin A mitogen response of peripheral blood mononuclear cells as the disease progressed.


A recombinant feline immunodeficiency virus envelope fusion protein stimulates peripheral blood lymphocytes from naive cats to proliferate in vitro. A region of feline immunodeficiency virus (FIV)/Glasgow-8 external envelope glycoprotein (env) incorporating the third and fourth variable regions (V3/V4) was cloned, inserted into the pGEX vector and expressed in Escherichia coli to yield milligram quantities of the recombinant polypeptide as a fusion protein with glutathione S-transferase. The fusion protein V3/V4GST was used in lymphocyte proliferation assays, where it consistently caused peripheral blood lymphocytes from naive cats to proliferate in a dose-dependent manner. Other FIV fusion proteins produced under identical conditions (V5GST and p24GST) and glutathione S-transferase alone did not cause proliferation in this system. The monoclonal antibody vpg15, which has been shown to block infection of susceptible cells in vitro, did not decrease the response to V3/V4GST. Human peripheral blood lymphocytes did not proliferate in response to V3/V4GST.


Evaluation of 9-(2-phosphonylmethoxyethyl) adenine therapy for feline immunodeficiency virus using a quantitative polymerase chain reaction. To determine the efficacy of 9-(2-phosphonylmethoxyethyl)adenine (PMEA) as a prophylactic chemotherapeutic agent for the treatment of lentivirus infections, three groups of specific pathogen free cats were treated with 0, 3, or 6 mg kg-1 twice daily doses of PMEA beginning 24 h prior to virus challenge with feline immunodeficiency virus Petaluma strain. Treatment was continued for 7 weeks post challenge. During this time cats were monitored for drug toxicity, virus specific antibody response, circulating viral antigen and infectious recoverable virus. To determine the long-term influence of PMEA therapy the cats were monitored for 1 year following the cessation of treatment. The low levels of infectious virus present in blood prompted the development of quantitative polymerase chain reaction assay to enumerate viral DNA burdens in the peripheral blood mononuclear cells of the infected cats and thereby assess drug efficacy. The results indicate that, although prophylactic PMEA did not prevent infection, it did substantially limit feline immunodeficiency virus replication. Furthermore, viral DNA levels remained low in the cats receiving drug a full year (the duration of the study) after cessation of treatment.


Pilot study of the effect of acemannan in cats infected with feline immunodeficiency virus. Acemannan, a complex carbohydrate shown to stimulate interleukin-1, tumor necrosis factor alpha and prostaglandin E2 production by macrophages, has also demonstrated antiviral activity in vitro against human immunodeficiency virus, Newcastle disease virus and influenza virus. A pilot study was undertaken to determine acemannan's effect in 49 feline immunodeficiency virus (FIV) infected cats with clinical signs of disease (Stage 3, 4 or 5), 23 of which had severe lymphopenia. Cats received acemannan either by intravenous (Group 1) or subcutaneous (Group 2) injection once weekly for 12 weeks, or by daily oral (Group 3) administration for 12 weeks. Upon entry into the study, cats were randomly assigned to one of the three groups. Laboratory analyses were performed at the beginning of the study and at Weeks 6 and 12. Cats were allowed to continue with a predetermined maintenance regimen of acemannan after completing the 12-week study. Thirteen cats died during the course of treatment. Upon necropsy, the most frequent histopathologic findings were neoplastic, kidney and pancreatic disease. Friedman's two-way ANOVA test showed no significant differences in efficacy among groups administered acemannan by the different routes. Therefore, groups were combined and a signed-ranks test was used to determine changes over time. A significant increase was seen in lymphocyte counts (P < 0.001). Neutrophil counts decreased significantly (P = 0.007), as did incidence of sepsis (P = 0.008). When cats entering with lymphopenia were analyzed separately, a much greater increase in lymphocyte counts was noted (235%) compared with non-lymphopenic cats (42%). A survival rate of 75% was found for all three groups. Thirty-six of 49 animals are alive 5-19 months post-entry. These results suggest that acemannan therapy may be of significant benefit in FIV-infected cats exhibiting clinical signs of disease.


Enhancement after feline immunodeficiency virus vaccination. Cats were vaccinated with one of the three preparations: purified feline immunodeficiency virus (FIV) incorporated into immune stimulating complexes (ISCOMs), recombinant FIV p24 ISCOMs, or a fixed, inactivated cell vaccine in quil A. Cats inoculated with the FIV ISCOMs or the recombinant p24 ISCOMs developed high titres of antibodies against the core protein p24 but had no detectable antibodies against the env protein gp120 or virus neutralising antibodies. In contrast, all of the cats inoculated with the fixed, inactivated cell vaccine developed anti-env antibodies and four of five had detectable levels of neutralising antibody. However, none of the vaccinated cats were protected from infection after intraperitoneal challenge with 20 infectious units of FIV. Indeed there appeared to be enhancement of infection after vaccination as the vaccinated cats become viraemic sooner than the unvaccinated controls, and 100% of the vaccinated cats became viraemic compared with 78% of the controls. The mechanism responsible for this enhancement remains unknown.


Retrovirus infections in non-domestic felids: serological studies and attempts to isolate a lentivirus. An African lioness from the Zoo of Zurich had to be euthanized because of an inoperable tumor. The serum tested negative for feline leukemia virus (FeLV) p27 antigen by enzyme-linked immunosorbent assay (ELISA) but was strongly positive for feline immunodeficiency virus (FIV) antibodies by ELISA and Western blot. When her only offspring and mate were tested for FIV, high antibody titers to FIV were also found in their serum. Lymphocytes were prepared from these two lions on different occasions and co-cultivated with specific pathogen free (SPF) cat lymphocytes in the presence of concanavalin A and recombinant human interleukin-2 (IL-2) for 6 weeks. The cell culture supernatants tested negative for Mg(2+)-dependent reverse transcriptase and FIV p24 by a double antibody sandwich ELISA throughout the culture period. Whole blood and buffy coat cells collected from these two lions were transmitted by intraperitoneal injection into two SPF cats. The two cats did not seroconvert for a period of 11 months nor could reverse transcriptase activity and FIV p24 antigen be demonstrated in the supernatant of several lymphocyte cultures. To determine the importance of lentivirus infections in zoo-kept wild felids, 124 serum samples were obtained from African lions, Indian and Siberian tigers, snow leopards, panthers, cheetahs and other wild cats from nine European zoos. In addition, serum samples collected from 12 Asiatic lions originating from Gir forest in the Indian State of Gujarat were included in this study. The sera were tested for antibodies to FIV, FeLV and feline syncytium-forming virus (FeSFV) by ELISA and Western blot using the respective viruses after gradient purification. In addition, some of the sera were also tested for antibodies to equine infectious anemia virus (EIAV) and Visna-Maedi virus (VMV). Antibodies to FIV were found in 30/53 (57%) of African lions, one of 18 tigers and one of four panthers. All other sera including those collected from the 12 Asiatic lions were negative for FIV antibodies. Some of the FIV positive lion sera had high antibody titers producing strong bands on Western blot strips even in dilutions of >> 1:1000. The Western blot pattern of the lion sera differed from that of domestic cats in that primarily p24 and to a lesser degree p17 was recognized. Antibodies to FeSFV were found in 14 animals (seven with strong, seven with intermediate, reaction). No correlation was found between FIV and FeSFV infection. Antibodies to FeLV were found in two cheetahs which later turned out to have been vaccinated with Leukocell, a FeLV vaccine.(ABSTRACT TRUNCATED AT 400 WORDS)


Feline immunodeficiency virus neurotropism: evidence that astrocytes and microglia are the primary target cells. To investigate the neuropathogenesis of feline immunodeficiency virus (FIV) infection in vitro, we have utilized three populations of cultured feline neural cells (astrocytes, microglia, brain endothelium) to assess the relative susceptibility to FIV infection, ability to produce viral antigens, and effects of infection on cell survival. Astrocytes appeared to be the most susceptible to infection, followed by microglia, whereas brain endothelial cells were relatively resistant to infection. Astrocyte infection resulted in syncytium formation and cell death, while microglial cells remained persistently and productively infected, without obvious cytopathic effects. These results suggest that FIV entry into the central nervous system probably does not occur via infected endothelium and that both astrocytes and microglia are more likely target cells for the virus.


Clinical and pathological findings in feline immunodeficiency virus experimental infection. A study is described of the clinical and pathological findings in 20 specific pathogen free cats infected when 1 year old with feline immunodeficiency virus and monitored over 12 months. Cats were divided into two groups (A and B). The clinical and clinicopathological features were studied in Group A. In Group B, at 1, 2, 4, 9 and 12 months post infection two cats were necropsied. Clinically all cats developed generalised lymphadenopathy, six cats were neutropenic and five cats lymphopenic. Three cats became febrile with conjunctivitis and anterior uveitis and one of these cats ultimately developed jaundice. Postmortem examinations confirmed a generalised lymphadenopathy involving peripheral and visceral lymph nodes with concurrent stimulation of splenic white matter and mucosal lymphoid tissue of the digestive tract and conjunctiva. Within the lymph nodes there was a reactive follicular hyperplasia accompanied by a paracortical hyperplasia with an increased paracortical vascularity. Unusual features were the presence of lymphoid follicles in the bone marrow, thymus and parathyroid tissue. In addition, aggregates of lymphoid cells were found within salivary glands, kidneys, sclera and choroid of the eye. One cat developed a lymphosarcoma affecting the liver and kidneys at 36 weeks post infection. The cat with jaundice had a cholangitis with marked biliary epithelial hyperplasia.


Detection of feline immunodeficiency virus infection in bone marrow of cats. Natural or experimental feline immunodeficiency virus (FIV) infection in cats is often associated with hematologic abnormalities which are similar to those observed in human immunodeficiency virus (HIV) infected patients. To determine if cells in bone marrow are infected with FIV and whether severity of hematopoietic disorder is correlated with the level of viral infection, bone marrow tissues from ten experimentally and two naturally FIV infected cats were examined by in situ hybridization for presence of FIV RNA. Seven of the 12 FIV infected cats were also naturally or experimentally coinfected with feline leukemia virus (FeLV). FIV RNA was detected mainly in megakaryocytes and unidentified mononuclear cells in the bone marrow of cats that were sick and had marrow hypercellularity and immaturity. These included all cats in the acute phase of FIV infection and two of seven long term FIV infected cats. One long term FIV infected cat with lymphosarcoma was also positive for FIV RNA in bone marrow cells. The other four long term FIV infected cats were relatively healthy, with normal bone marrow morphology, and were negative for FIV infected cells. Bone marrow from three non-infected and two cats infected with FeLV alone were also negative for FIV RNA by in situ hybridization. We concluded that megakaryocytes and mononuclear cells were targets of the viral infection and that the presence of FIV RNA in cells of the bone marrow correlated with marrow hypercellularity and immaturity, and severity of illness.


Decreased mitogen responsiveness and elevated tumor necrosis factor production in cats shortly after feline immunodeficiency virus infection. We present the results of an investigation into the effects of feline immunodeficiency virus (FIV) infection on the response to mitogens and cytokine production in the first month of infection. We were able to demonstrate a depression of response of peripheral blood mononuclear cells to the mitogens concanavalin A, phytohaemagglutinin and pokeweed mitogen, with the response to pokeweed mitogen being most severely affected. The response of the cells of the spleen were affected by 10 days post infection and these could not be augmented by the addition of exogenous interleukin-2 (IL-2). The response of mesenteric lymph node cells was not affected until 20 days post infection and this could be partially restored by the addition of exogenous IL-2. IL-2 production was unaffected in peripheral blood mononuclear cells, slightly depressed in mesenteric lymph node cells and slightly elevated in spleen cells. Tumor necrosis factor levels were significantly elevated with respect to controls within 10 days of infection. These studies suggest that there are a number of changes in the immune response of FIV infected cats early in infection and this may determine the subsequent outcome of the infection.


Tumor necrosis factor alpha levels in cats experimentally infected with feline immunodeficiency virus: effects of immunization and feline leukemia virus infection. Tumor necrosis factor alpha (TNF alpha) levels were determined by enzyme-linked immunosorbent assay (ELISA) and by cell culture bioassay in supernatants of lipopolysaccharide-stimulated feline monocyte cultures and in cat serum samples. There was a good correlation between the results obtained by the two methods. From the fact that TNF alpha was neutralized quantitatively by antibodies to human TNF alpha in feline monocyte supernatants and in feline sera, it was concluded that feline TNF alpha immunologically cross-reacts with human TNF alpha and that the human TNF alpha ELISA can be used to quantitate feline TNF alpha. During the first 6 months after experimental feline immunodeficiency virus (FIV) infection no differences in serum TNF alpha values were observed between infected and non-infected cats. TNF alpha levels increased significantly after primary vaccination with a feline leukemia virus (FeLV) vaccine in FIV infected cats over those in the non-infected controls. During secondary immune response TNF alpha levels rose transiently for a period of a few days in both the FIV positive and the FIV negative cats. After FeLV challenge, TNF alpha levels increased in all animals challenged with virulent FeLV for a period of 3 weeks. This period corresponded to the time necessary to develop persistent FeLV viremia in the control cats. It was concluded from these experiments that in the asymptomatic phase of FIV infection no increased levels of TNF alpha are present, similar to the situation in asymptomatic HIV infected humans. Activation of monocytes/macrophages in FIV infected cats by stimuli such as vaccination or FeLV challenge readily leads to increased levels of TNF alpha.


Transmission of feline immunodeficiency virus from infected queens to kittens. This study demonstrates the transmission of feline immunodeficiency virus (FIV) from infected queens to kittens in two separate litters. Queen 1 was infected by intravenous administration of FIV at 22 days prior to parturition. Two out of three kittens from the litter were found to be viremic at 10 weeks of age as detected by culture isolation and polymerase chain reaction detection of FIV DNA in peripheral blood mononuclear leukocytes. The third kitten remained aviremic through 40 weeks of age. Queen 2 was infected by subcutaneous administration of FIV 2 days prior to parturition. This litter also had two out of three kittens infected with FIV; however, viremia was not detected in one of the kittens until 21 weeks of age. Culture isolation was found to be superior to polymerase chain reaction for the early detection of FIV, and viremia was found to precede seroconversion by up to 4 weeks. Although all infected kittens have remained healthy, depressed CD4:CD8 lymphocyte ratios suggest that clinical disease may develop. This study suggests that FIV infection in cats may be a useful model system for the study of HIV transmission from mothers to infants.


[The legal and ethical aspects of nerve tissue transplantation]. The authors have specified the following criteria for the withdrawal of embryonal tissue at their department: 1) only tissue from dead fetus is allowed to be used in neurotransplantation; 2) embryonal tissue is to be obtained after spontaneous abortions from volunteers or from women asking for artificial abortion; 3) the women should be informed about the curative purposes of embryonal tissue voluntary donorship and they must give a written consent; 4) decision on abortion should be separated from the use of embryonal tissue; 5) women should not know recipients; no payments should be made for tissue; 6) the donor is not permitted to impregnate in order to use embryos for research or clinical purposes; 7) sampling of BWR, HBsAG, anti-HIV, cytomegalovirus, herpes I and II is to be made for serologic examinations and that from the cervix for cultivation and sensitivity, as well as ultrasound verification of a germinal age is done in potential donors; 8) consent should be signed to embryonal brain transplantation by recipient or his legitimate deputy if the recipient is certifiable. The above criteria should protect both the donor and the recipient. The use of embryonal tissue cultures seems to be promising. In addition to legal and ethic problems, immunological problems and problems concerning the aseptic withdrawal of embryonal tissue are falling off.


[The pathogenesis of pulmonary complications in severe craniocerebral trauma]. The paper summaries authors' experience with lifetime and postmortem pulmonary changes in 223 victims with severe craniocerebral injury, as well as the data available in literature on this point. The investigations have shown that microcirculatory disorders make a great contribution to the development of pulmonary complications and revealed specific feature of the natural history of the shock lung in patients with severe craniocerebral injury. The paper provides strong evidence for the neurotrophic pattern of pulmonary complications and proposes a scheme of their development in this injury.


[The corrective effect of perfluorocarbon emulsions on the cerebral hemodynamics in craniocerebral trauma]. The article discusses the results of study of the effect of perfluorocarbons on the cerebral blood flow and metabolism of an intact brain and in severe graded craniocerebral injury in experiments on 25 dogs. The results make it possible to prognosticate the efficacy of using preparations based on emulsions of organoperfluorine compounds in neuroreanimatology to correct disorders of cerebral blood flow and metabolism in the foci of brain contusion.


[The adaptive-compensatory reactions of the systemic circulation in the acute period of severe craniocerebral trauma]. The paper deals with the most important problem of performance of central hemodynamics in the acute period of craniocerebral injury. The author recorded an informative set of central hemodynamic parameters, which allowed him to assess not only central hemodynamics, but its reserve and compensatory potentialities. He investigated variations of the parameter set in various brain injuries, by using it as a functional test for some pharmacological agents, which is essential in evaluating their efficacy in these abnormalities. The findings define the role of disorders of blood circulation regulation and cardiac performance. The mode of assessing the hemodynamic parameters is promising in obtaining new data on this point.


[Changes in the hormonal balance and erythropoiesis following neurosurgical operations]. The work analyses the relationship between hormonal disorders in the postoperative period and the dynamics of postoperative anemias in patients who were operated on for brain tumors of various localization. It was found that hormonal changes have an effect on the process of hematopoiesis manifested by inhibition of erythroid differentiation in the bone marrow. The principal role in this case belongs to hypercortisolemia which is intensified by postoperative hormone therapy.


[The clinical picture and differential diagnosis of rhinosinogenic brain abscesses]. Some specific features of the course of rhinosinogenic brain abscesses (RBA) in adults and children were distinguished on the basis of analysis of the clinical data in 49 patients. RBA is encountered more frequently among adults and rarely among children, especially those of young age. In adults RBA are mostly consequent upon chronic frontitis or polysinusitis, while in children they occur after maxillary sinusitis, and in young children also after acute purulent processes in the nasal cavity. The contact route of infection of the cranial cavity and solitary abscesses are usually encountered in adults, and the hematogenous-metastatic route in children with the formation of solitary RBA and, in many cases, of multiple and multichamber abscesses. In children the abscesses grow to a large size. The hypertension syndrome is clearly defined in adults, while the hydrocephalus-hypertension syndrome comes to the forefront in children. The infectious-toxic symptoms are more marked in children. Focal neurologic symptomatology is manifested more markedly in adults and is less marked and labile in children.


[The diagnosis of fractures of the anterior cranial fossa in craniofacial trauma]. The authors examined 436 patients with acute trauma of the upper and middle zones of the visceral cranium. Injury to the anterior cranial fossa was discovered in fractures of the orbitonasal complex, isolated fractures of the upper zone, fractures of the Le Fort II and Le Fort III types, and multiple fractures of the upper and middle zones of the face. Such fractures were found in 158 cases, in 76 of them (48.1%) damage to the anterior cranial fossa was recognized. A relationship was found between the frequency of fractures of the anterior cranial fossa and the type of injury to the bones of the face. The features of injury to the separate structures of the anterior cranial fossa are specified. The modified program of panoramic sonography by means of a Zonark apparatus makes it possible to examine the anterior cranial fossa of a patient in a supine position with a small number of exposures, which makes the examination much easier for the patient.


[Selective epidural phlebography in the diagnosis of tumors of the cervical spinal cord and of the cauda equina roots]. The method of selective epidural phlebography (SEP) was applied in the diagnosis of tumors of the cervical segment of the spinal cord and the roots of cauda equina in 45 patients. The diagnosis was confirmed in 32 of the 37 patients who were operated on. The SEP method does not produce serious complications and allows circulatory disorders in the vertebral venous plexuses to be revealed. The diagnostic value of the method is higher in tumors of superocervical localization and hourglass tumors.


[The angiographic diagnosis of vascular spasm in ruptured cerebral aneurysms]. The authors compared the results of qualitative and quantitative appraisal of arterial spasm according to the angiographic findings in 85 patients with the severity of their condition and neurological symptomatology in the acute period of rupture of a brain aneurysm. The spasm was diagnosed by quantitative appraisal in 80% and by qualitative appraisal in 72.9% of cases. Appraisal of the spasm in various periods after rupture of the aneurysm showed it to be most intense on the 9th-10th day. The vascular spasm (VS) was the main cause of the neurological symptomatology in the acute period of rupture of the aneurysm. Constriction of the arteries to 1.65 in quantitative appraisal of the spasm and its extension to 3-4 segments and more in qualitative appraisal were attended by diffused consciousness and stem and focal hemispheric symptomatology. Simultaneous use of both methods for VS appraisal makes it possible to evaluate its extension and the degree of arterial constriction and distinguish the most severe forms of spasm.


Inclusion body myositis with abundant ring fibers. A 37-year-old male presenting with a 7-year history of leg weakness was found to have moderate weakness confined to the quadriceps and myopathic changes on electromyography. Serum creatine phosphokinase was 2130 units/liter. Biopsy of the quadriceps muscle revealed an inflammatory myopathy with cytoplasmic and nuclear filamentous inclusions characteristic of inclusion body myositis. An unusual finding was the large proportion of ring fibers along with severe atrophy and fibrosis. The pathogenesis of the ring fibers in this setting is discussed.


"Reducing body"-like inclusions in skeletal muscle in childhood-onset acid maltase deficiency. Unusual inclusions with some of the features of "reducing bodies" were encountered in the skeletal muscle biopsy of a 2.5-year-old boy with childhood-onset acid maltase deficiency. The biopsy revealed a vacuolar myopathy with lysosomal storage of glycogen and eosinophilic refractile inclusions in myofibers, which appeared dark blue with the menadione-nitroblue tetrazolium reaction. The significance of the association of inclusions with reducing properties in the setting of acid maltase deficiency is discussed.


[A nontraditional approach to anesthesiological support and intensive therapy in neurosurgery]. Using mediator agents in definite combinations allows one to simulate the functions of the genetically determined levels of adaptive reaction integration. The action of combinations of narcotic analgetics with varying receptor affinity and alpha 2-adrenomimetics impoverishes and modulates a flow of sensory information to the higher centers of autonomic integration and largely relieves efferent impulsation. This ensures harmonicity of the reactions and makes for the operation of local sanogenetic mechanisms of the brain at functional rest. Anesthesia performed in the early postoperative period by using a combination of the narcotic analgetic phentanyl and the centrally acting alpha 2-adrenoagonist clofelin enables the authors to significantly improve the outcomes of treatment.


[Magnetic resonance tomography in spinal cord trauma]. A total of 44 patients with sequelae of spinal injuries in the periods of 2 months to 46 years were studied by X-ray, neurological and NMR techniques and divided into 3 groups: (1) those with open injuries, (2) those with closed injuries concurrent with compression of the spinal marrow and/or roots of the cauda equina; 3) those with closed injuries without compression. It was shown that NMR tomography enabled the status of the vertebral canal and spinal marrow to be assessed and indications for surgical interventions to be specified. Spinal compression may in later periods result in post-traumatic progressive myelopathy appeared as atrophy of the spinal marrow and myelomalacia (with cyst formation in 18% of cases). The syndrome of progressive myelopathy does not generally develop in patients without spinal compression.


[The assessment of brain stem function in the acute period of aneurysm rupture]. The functional status of the brain stem was assessed by recording evoked stem auditory potentials (ESAP) in 76 patients in the acute period of arterial aneurysm rupture. Abnormalities of ESAP were detected in 39.4% of the patients. The determinant of ESAP disorder in aneurysm rupture is the occlusive-dislocation syndrome caused by intracranial hematoma, more than 50 ml in volume, ventricular tamponade or extensive vascular spasm.


Distribution of cellular carbohydrate moieties in human dysontogenetic brain tumors, especially in craniopharyngioma and epidermoid/dermoid. Cellular carbohydrate moieties of 65 human dysonotogenetic brain tumors (craniopharyngioma, epidermoid/dermoid, Rathke cleft cyst, germinoma and non-germinomatous germ-cell tumors) and 60 common brain tumors (glioma, meningioma, neurinoma and pituitary adenoma) were investigated histochemically using sections from Ulex europaeus (UEA-1), Dolichos biflorus (DBA), peanut (PNA) and soybean (SBA), and with anti-blood group A and LewisY (LeyY) antibodies. In craniopharyngiomas and epidermoid/dermoids, it was found that PNA and SBA binding sites existed in suprabasal cells of the epithelium, and that antigen of either blood group A or H (demonstrable by UEA-1) existed in more differentiated epithelial cells compared to the results reported in normal human skin epidermis. Rathke cleft cysts were stained with PNA or SBA, and two out of three Rathke cleft cysts also expressed either H or A antigen. In addition, DBA binding sites, as well as LeY antigen, were frequently seen in craniopharyngiomas and Rathke cleft cysts, but they were entirely absent in the epithelium of epidermoid/dermoid. On the other hand, PNA and SBA reactivities was also found in common brain tumors, while blood group A, H and LeY antigens and DBA reactivity were almost absent in these tumors. These findings demonstrate that carbohydrate moieties such as those of blood group antigens reported to be found in human skin epidermis exist in a similar form in craniopharyngioma, epidermoid/dermoid and the Rathke cleft cyst. The identification of blood group A, H and LeY antigens and DBA reactivity in brain tumors seems to be considerably limited and specific.(ABSTRACT TRUNCATED AT 250 WORDS)


[The noninvasive diagnosis of cerebral arteriovenous aneurysms by transcranial dopplerography]. An absolute increase of blood velocity in the arteries of brain arteriovenous aneurysms is of the greatest importance in their diagnosis by transcranial Doppler ultrasonography. If the branches of brain great vessels are involved in blood supply of arteriovenous aneurysms, there is an increase in blood velocity only in the initial portions of the great vessels up to the origin of these branches. A relative increase in brain great vasculature blood velocity is of great diagnostic value. Arteriovenous aneurysms whose blood supply is provided by the anterior cerebral artery are typified by higher blood velocity in the anterior cerebral artery than that in the middle cerebral artery, whereas the reverse is true for health.


Urinary LTE4 excretion in antigen-provoked asthmatic patients treated with the inhaled LTD4 antagonist, L-648,051. Leukotriene (LT) E4 represents the major LT metabolite in man, and its urinary excretion can be used as an indirect marker of systemic LTC4 and/or LTD4 synthesis and release. In the present study LTE4 excretion was monitored for 24 h in 12 atopic patients with mild asthma undergoing antigen bronchoprovocation as part of a double-blind, placebo-controlled, two-period cross-over study of the aerosol-delivered LTD4 antagonist, L-648,051. Urinary LTE4 excretion was also studied separately in six of the patients after inhaling only diluent. Urine was sampled before, and serially after antigen challenge, at intervals corresponding to the immediate (0-3 h postchallenge) and late (3-6, 6-12, 12-24 h postchallenge) asthmatic reactions. LTE4 was determined by reversed-phase HPLC and radioimmunoassay. Forced expiratory volume in 1 s (FEV1) was recorded serially through 8 h after inhalation of antigen and diluent. Compared to base-line measurements, antigen bronchoprovocation induced significant increases in mean LTE4 excretion rates 0-3 h postchallenge (i.e. during the immediate asthmatic response) after treatment with both placebo (P < 0.01) and L-648,051 (P < 0.05). These mean LTE4 excretion rates in the immediate phase were also significantly higher than the mean rates in the late phase (3-6 h and beyond); the excretion rates of LTE4 at these later time intervals were similar to base-line values. After inhalation of diluent, the LTE4 excretion rates in the intervals 0-3, 3-6, 6-12 and 12-24 h were unchanged from base-line values.(ABSTRACT TRUNCATED AT 250 WORDS)


Leukotriene B4 and platelet-activating factor in human skin. Acute inflammatory reactions are characterized by leukocyte infiltration associated with increases in vascular permeability and in local blood flow. Leukocyte infiltration can be induced by chemotactic factors such as leukotriene B4 (LTB4) and paf-acether (formerly known as platelet-activating factor) that can be generated within inflammatory lesions. Vascular permeability and increase in blood flow are also affected by LTB4 and paf-acether, as well as by several other substances, including histamine and prostaglandins. Derived from arachidonic acid via the 5 lipo-oxygenase pathway, LTB4 is one of the most potent leukocyte chemotactic substances known. Intradermal injections of LTB4 induce dermal neutrophil infiltration in animal models and in humans. Topical application of LTB4 to human skin induces intraepidermal micro-abscesses containing numerous intact neutrophils. LTB4 has been found to be increased in psoriatic lesions, but its synthesis by epidermal cells remains undecided. Like other leukotrienes, LTB4 can stimulate DNA synthesis in cultured human epidermal keratinocytes. However, receptors for LTC4 but not for LTB4 have been found on human keratinocytes in culture. Paf-acether is an ether-linked phospholipid identified as 1-O-alkyl-2-O-acetyl-sn-glycero-3-phosphocholine and is considered to be one of the most potent mediators of acute allergic and inflammatory reactions. For instance, intradermal injection of paf-acether induces inflammatory events such as neutrophil infiltration and increase in vascular permeability. Recent data suggest that cutaneous cells, such as fibroblasts and keratinocytes, are capable of producing paf and that paf is released during the development of allergic cutaneous reactions.(ABSTRACT TRUNCATED AT 250 WORDS)


Vitamin D3 and skin diseases.

Vitamin D3 and skin diseases. The physiologically active metabolite of vitamin D3, 1 alpha,25-dihydroxycolicalciferol [1,25(OH)2D3, calcitriol] has achieved the status of a hormone. It is believed to mediate its effects by binding to a specific receptor which belongs to the family of nuclear receptors for glucocorticoids, estrogens, thyroxine, and retinoid acid. It has been discovered that 1,25(OH)2D3 has the ability to regulate growth and differentiation in many cell types, including cancer cells, epidermal keratinocytes, and activated lymphocytes. This has set the stage for the development of a new class of compounds with potential usefulness in hyperproliferative and immune-mediated diseases. Ideally, such agents should possess potent effects as regulators of cell proliferation and differentiation at concentrations well below those that may induce side effects related to the classical vitamin D activity on calcium absorption and bone mineralization. In addition to 1,25(OH)2D3, the synthetic vitamin D3 analogues 1 alpha-OH-D3, 1,24(OH)2D3, and calcipotriol have undergone clinical evaluation. Calcipotriol has been studied most extensively. Compared with 1,25(OH)2D3, calcipotriol is about 200 times less potent in its effect on calcium metabolism although similar in receptor affinity. In double-blind, placebo-controlled multicenter studies, topical calcipotriol has been shown to be both efficacious and safe for the short- and long-term treatment of plaque-type psoriasis. Because some of the novel vitamin D analogues are potent regulators of cell growth and immune responses, they may be of potential interest in the treatment of ichthyoses, cancer, and autoimmune diseases.


Ménière's disease and antibody reactivity to herpes simplex virus type 1 polypeptides. It has been reported that the inner ear is capable of responding to antigen challenge. In this study, we have investigated the antibody reactivity to herpes simplex virus 1 (HSV-1) proteins in sera from patients with Ménière's disease. Serum from 21 patients scheduled to undergo endolymphatic sac decompression for Ménière's disease was obtained. The sera from 21 age- and sex-matched individuals without a history of ear disease served as the control. An enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies to HSV-1, HSV-2 cytomegalovirus (CMV), varicella-zoster virus (VZV), and measles virus. Immunoblotting was used to confirm and further evaluate the HSV-1 antibody response. All but one patient had antibodies to HSV-1. ELISA mean log titers were significantly lower in the patient group to CMV and VZV when compared with controls. A pattern of generally higher antibody reactivity in patients with Ménière's disease was demonstrated to the individual SDS-PAGE HSV-1 polypeptides as judged by immunoblotting. The HSV-1 antibody response found in patients with Ménière's disease may indicate viral reactivation and denotes the importance of further studies on the role of infectious agents in this disease.


[Coronary disease of transplanted heart. Physiopathology, prevention]. Coronary artery disease in cardiac transplant recipients is the main cause of mortality after the first postoperative year. This atheroma has unique anatomical features with widespread infiltration of the intima by inflammatory cells. The different physiopathological hypotheses are analysed. Immunological processes are probably responsible, suggesting chronic rejection. The cytomegalovirus could be an inductive mechanism. The classical vascular risk factors probably play a role but their action is not as clear as that of immunological and viral factors. Hyperlipidaemia is a causative mechanism and the predisposing role of steroid therapy is also recognised. From the practical point of view, correction of the classical risk factors is the only available therapeutic possibility at present.


A theoretical study of the mechanism of oxygen binding by model anthraquinones. I: Quantum mechanical evaluation of the oxygen-binding sites of 1,4-hydroquinone. The undesirable cardiotoxicity of some important classes of antitumor drugs, such as anthracycline derivatives, is caused by their mediation of the one-electron reduction processes of the oxygen molecule which produces the highly toxic superoxide anion radical. Recent studies enable the conclusion to be drawn that the first and rate-limiting stage of this process is the formation of complexes of the drug anthraquinone moiety with 1 delta g molecular oxygen. The complexes can easily undergo one-electron reduction, whose product dissociates into the unchanged drug molecule and the superoxide anion radical. The present study reports quantum mechanical calculations of the structure and the energies of the possible oxygen complexes of the most simplified model compound: 1,4-benzenediol (1,4-hydroquinone); 2,3-dihydro-2,3-epidioxy-, 2,5-dihydro-2,5-epidioxy- and 1,4-epidioxy-1,4-benzenediol (the 2,3-, 2,5- and 1,4-peroxide). Calculations were carried out with the use of ab initio (STO-3G, 4-31G, and 6-31G) and semiempirical MNDO methods with total geometry optimization. The optimized geometry parameters were found to be in a reasonable agreement with the available crystal data. During the oxygen complex formation with hydroquinone, charge transfer occurs from hydroquinone to the oxygen molecule. Supplementary MNDO calculations have shown that the stability of 2,3-peroxide is increased substantially upon the ionization of one of the hydroxyl groups of hydroquinone prior to oxygen binding, which increases the electron density of the benzene ring. These findings result in a prediction that the anthracycline derivatives with electron-withdrawing substituents in the II-electron moiety should exhibit diminished affinity towards oxygen and, consequently, diminished ability to peroxidation. It has also been found that the relative energies of different peroxides are well represented even in the STO-3G ab initio calculations which will enable the further extension of the study to the complete II-electron moiety of the actual anthracycline derivatives.


An adriamycin-resistant subline is more sensitive than the parent human small cell lung cancer cell line to lonidamine. Lonidamine, a unique new type of anticancer agent, has been shown to exert marginal activity in patients with resistant small cell lung cancer. We have assessed cytotoxic activity using seven human small cell lung cancer cell lines and the drug resistant small cell lung cancer sublines, Adriamycin-resistant SBC-3/ADM100, etoposide-resistant SBC-3/ETP and cisplatin-resistant SBC-3/CDDP. Although lonidamine had only a minimal activity in seven human small cell lung cancer cell lines, SBC-3/ADM100 cells were more sensitive to lonidamine than the parent SBC-3 cells to a 1.6-fold extent in terms of IC50. SBC-3/CDDP and SBC-3/ETP had no cross-resistance to lonidamine at all. These observations may be of potential clinical significance in treating small cell lung cancer.


Post-train depression of ganglionic transmission in the presence of d-amphetamine. Repetitive stimulation (30 Hz, 20 sec) of the preganglionic nerve to the hamster isolated stellate ganglion in a control solution had only small effects on the compound action potentials of the postganglionic nerve (0.2 Hz) after the train. When 10(-5) M d-amphetamine was added to the superfusing solution, the repetitive stimulation induced a post-train depression of the postganglionic compound action potential. The mean reduction of the compound action potential during the post-train depression in d-amphetamine was 37 +/- 15% and the duration was from 60 to 120 sec. Post-train depression only occurred after long conditioning trains and was never observed with less than 100 stimuli in the conditioning train. A post-train depression (45 +/- 8%) was also observed in the presence of 10(-5) M cocaine. The post-train depression in d-amphetamine was significantly reduced by the alpha-adrenoceptor antagonist, phentolamine (10(-5) M) and by the alpha 2-adrenoceptor antagonist, yohimbine (10(-6) M), but not by the alpha 1-adrenoceptor antagonist, prazosin (10(-5) M). There was no post-train depression in the presence of d-amphetamine in ganglia from five hamsters that were pretreated with reserpine (6 mg/kg, i.p., 4 hr). Atropine (10(-6) M) increased and prolonged the post-train depression in the presence of d-amphetamine. These results indicate that repetitive stimulation in the presence of d-amphetamine leads to a post-train depression of the postganglionic compound action potential that is mediated by an action of endogenous catecholamines at alpha 2-adrenoceptors. Muscarinic cholinoceptors do not appear to be involved in the generation of the post-train depression.


Further examination of 9-alkyl- and sugar-modified anthracyclines in the circumvention of multidrug resistance. Anthracyclines possessing either a 9-alkyl modification in the A-ring of the tetracyclic aglycone and/or specific changes to the amino sugar moiety retain effective cytotoxic activity against multidrug resistant (MDR) cell lines. To obtain a better understanding of the structural features responsible for this potentially valuable behaviour, we used the MTT tetrazolium dye reduction assay to calculate resistance factors (RF = the ratio of ID50 for the drug-resistant line to that for the parental line) for the EMT6/P mouse mammary tumour and its MDR variant EMT6/AR1.0, and the H69/P human small cell lung cancer line and its MDR counterpart H69/LX4. Both MDR lines exhibit marked resistance to doxorubicin, MDR 1 gene amplification, hyperexpression of the membrane P-glycoprotein and reduced drug accumulation. RF values for doxorubicin were 34 and 131 in the EMT6 and H69 cell line pairs, respectively. The 9-alkyl-substituted anthracyclines were confirmed as having RF values 9- to 15-fold lower than those for doxorubicin. The 9-ethyl analogues Ro 31-1966 (RF for EMT6 2.2, RF for H69 4.7) and Ro 31-1749 (RF for EMT6 3.9, RF for H69 9.5) were superior to the previously studied 9-methyl analogue Ro 31-1215 (RF for EMT6 8.1 RF for H69 12.4). A clear trend for RF values to decrease with increasing 9-alkyl chain length was also noted in the structurally more complex aclacinomycin series. For example, 13-methyl-aclacinomycin (RF for EMT6 1.0, RF for H69 2.2) featuring a 9-isopropyl moiety was superior to the 9-alkyl-containing aclacinomycin A (RF for EMT6 4.7, RF for H69 5.8), and this was in turn more effective than the 9-methyl analogue sulfurmycin A (RF for EMT6 6.4, RF for H69 14.2). The trisaccharide moiety was not an essential feature for activity against MDR lines in the aclacinomycins, as shown by the low RF value with aklavine (RF for EMT6 2.1, RF for H69 2.5). However, a small change in one of the sugar moieties of aclacinomycin A, as in marcellomycin, resulted in a considerable increase in RF values (RF for EMT6 18.5, RF for H69 25.3). The complex anthracyclines AD 32 (RF for EMT6 6.5, RF for H69 11.7) and particularly tetrahydropyranyl-doxorubicin (RF for EMT6 1.4, RF for H69 3.2) were effective against MDR lines.(ABSTRACT TRUNCATED AT 400 WORDS)


Effects in vitro of two marine substances, chlorolissoclimide and dichlorolissoclimide, on a non-small-cell bronchopulmonary carcinoma line (NSCLC-N6). The antiproliferative activity of two nitrogenous labdane cytotoxic substances from Lissoclinum voeltzkowi Michaelson (Urochordata), dichlorolissoclimide (P2) and chlorolissoclimide (P1), was studied in vitro on a continuous human non-small-cell bronchopulmonary carcinoma line (NSCLC-N6) at the cell cycle level. This antiproliferative effect resulted from a blockade of G1 phase cells. Mortality occurred, regardless of the degree of cell ploidy, with cell transition to an out-of-cycle situation characteristic of a G1D terminal maturation state.


Different effect of dGTP on 2'-deoxyadenosine metabolism in mitochondria and cytosol. Two enzymes participating in 2'-deoxyadenosine (dAdo) metabolism: dAdo kinase (dAdoK EC 2.7.1.76) and adenosine deaminase (ADA, EC 3.5.4.4) were partially purified from rat liver mitochondria and cytosol and influence of nucleosides and nucleotides on the activity of these enzymes were investigated. Mitochondrial and cytosol dAdoK are separate proteins, while ADA from both subcellular fractions possesses similar physical properties. dGTP, a competitive inhibitor of mitochondrial dAdoK, inhibits cytosol ADA in a mixed way but activates mitochondrial ADA and cytosol dAdoK. A possible effect of dGTP on dAdo metabolism in mitochondria and cytosol is discussed.


Mechanisms of action of sodium channel blocker: gate-related receptor hypothesis. A new hypothesis on mechanisms of action of the sodium channel blocker, the gate-related receptor hypothesis, has been proposed in this paper. Furthermore, a mathematic model and parameter estimation procedure for the kinetics analysis have also been given. Using the model and procedure, modeling and analyzing of cardiac sodium channel blockade by lidocaine and dauricine have been performed. The model-estimated results were not only coincident with those documented, but also provided some new information which may be helpful for understanding mechanism of blocking action of both drugs on sodium channels. The studies suggested that the gate-related receptor hypothesis is good for the elucidation of mechanisms of action of the sodium channel blocker.


Immunosuppression by human seminal plasma--extracellular organelles (prostasomes) modulate activity of phagocytic cells. Prostasomes are trilamellar to multilamellar vesicles produced by the acinar cells of the human prostate and are present in appreciable amounts in normal human semen. The aim of this work was to study the effect of prostasomes on human polymorphonuclear cell and monocyte function. Functional activity of human neutrophils and monocytes was studied after incubation with prostasomes isolated from normal human seminal plasma. The following functional tests were employed: ability to ingest latex particles and opsonized bacteria (St. aureus) and ability to generate superoxide anion in response to PMA and FMLP. The latter was determined by measuring the superoxide-dismutase inhibitable reduction of ferricytochrome c to ferrocytochrome c at 550 nm. Expression of cell surface markers and interactions of prostasomes with cells were studied by cytofluorimetry. We show that prostasomes bind rapidly to the leukocyte cell membrane followed by internalization of adsorbed material. Interactions of prostasomes with neutrophils and monocytes inhibits their ability to phagocytose latex particles. The ability to ingest opsonized bacteria is, however, not impaired. Our results also show that incubation of leukocytes with prostasomes effectively inhibits superoxide anion generation in response to activation by PMA and FMLP. Prostasomes may play a complementary role to other immunosuppressive factors contained in the human semen. They may protect sperm cells from deleterious effects of phagocytosing cells, prolong their life, and consequently enhance the chance of conception. At the same time prostasomes may have a permissive effect on sexually transmitted diseases.


Regulation of GABAA receptor channels by anticonvulsant and convulsant drugs and by phosphorylation. The GABAA receptor channel is a highly regulated receptor. The function of the receptor may be modified by drugs which alter the rates of binding of GABA, modify the gating of the channel or block the channel. It is also likely that phosphorylation of the receptor subunits modifies the biophysical properties, stability or assembly of the receptor. While GABAergic inhibition plays a major role in the regulation of neuronal excitability, a role for altered GABAergic inhibition in the pathogenesis of epilepsy remains to be proven. The demonstration that GABAA receptors are composed of multiple subunits and that the properties and pharmacology of GABAA receptors are different for different subunit combinations suggests that GABAA receptor heterogeneity may be of importance in determining the properties of GABAergic inhibition in different regions of the nervous system. While it is clear that GABAA receptor heterogeneity is present in the nervous system, a role for receptor heterogeneity in the pathogenesis of epilepsy remains uncertain. GABAA receptor heterogeneity may have implications for the treatment of epilepsy. It is quite possible that drugs which regulate GABAergic function may have variable efficacy in different regions of the nervous system due to expression of receptors with subunits that have different sensitivity to allosteric regulators. Furthermore, it is likely that there are developmental changes in the stoichiometry or subunit composition of GABAA receptors rendering the developing nervous system more or less sensitive to the effects of GABAergic anticonvulsant drugs. In addition to the heterogeneous expression of GABAA receptors, other issues concerning the regulation of GABAergic function are of potential importance. The regulatory events that control the expression of specific receptor subtypes and levels of GABA receptors are unknown. The post-translational events that regulate GABAA receptor function are uncertain. It is possible that post-translational regulation of GABAA receptors by phosphorylation may contribute to altered GABAA receptor function in epilepsy. To understand the role of GABAA receptor heterogeneity in the pathogenesis of epilepsy will require the combination of biophysical and molecular biological techniques. It will be important to determine not only whether the properties of GABAA receptors have been altered in a specific form of epilepsy, but also whether gene expression has been altered.


The effects of AMPA receptor antagonists on kindled seizures and on reflex epilepsy in rodents and primates. Two potent glutamate antagonists, NBQX and GYKI 52466, that act selectively on non-NMDA receptors, have been tested for anticonvulsant activity in 3 models of reflex epilepsy (sound-induced seizures in DBA/2 mice and in genetically epilepsy-prone rats and photically-induced myoclonus in Papio papio) and in amygdala kindled rats. Both compounds potently but transiently suppress reflexly-induced epileptic responses. GYKI 52466 also reduces behavioral seizures and afterdischarge duration in amygdala kindled rats, but with a lower potency than it suppresses reflex epilepsy. These data are similar to earlier results with antagonists acting selectively on NMDA receptors; they do not support a specific involvement of enhanced AMPA receptor sensitivity as a major factor in the expression of kindled seizures.


Interictal behavioral disturbances: a search for molecular substrates. Postictal symptoms can be disabling in themselves, but their underlying substrates may endure, giving rise to epilepsy-induced interictal behavioral disorders. Chronic temporal lobe epilepsy is reported to be associated with a variety of interictal behavioral changes which often take the form of affective disturbances. Depression, among the more common interictal psychological dysfunctions suffered by patients with temporal lobe seizures, could reflect epilepsy-induced alterations in normal opioid peptide mechanisms. In experimental animal models, certain postictal behaviors have been shown to be opioid-mediated. Furthermore, an experimental model of interictal behavioral disturbance resembles stimulation-induced defensive rage, which can be relieved by intracerebral administration of opioid peptides. Defensive rage is a species-specific behavior encountered in cats. Its correlate in humans would be difficult to predict in view of the stronger cortical control; however, it may manifest rather as insecurity, irritability, and perhaps depression. Extrapolation of animal experiments would suggest that depression and certain other common postictal and interictal affective disturbances seen in patients with temporal lobe seizures reflect mechanisms more related to opiate withdrawal, than to direct opiate actions. The activity-induced plasticity associated with recurrent temporal lobe seizures, therefore, should result in changes in opioid function that predispose to withdrawal phenomena. Limbic seizures induce enhanced enkephalin synthesis lasting for up to 2 weeks. Recurrent seizures in experimental animals, however, cause paradoxical up-regulation of mu opiate receptors. Patients with temporal lobe epilepsy demonstrate enhanced mu receptor binding in the neocortex of the epileptogenic temporal lobe on PET. The reasons for this enduring interictal effect are not clear. Nevertheless, if animals or patients become dependent on enhanced endogenous opioid activity as a result of seizures, and also have up-regulation of mu receptors, then severe withdrawal effects, such as defensive rage in cats or depression in humans, might be expected when seizures do not recur frequently. Plotting the time course of mRNAenk and enkephalin expression after seizures, and the time course of symptoms of interictal behavioral disturbances, may demonstrate a temporal relationship that supports this hypothesis. For instance, depression or other withdrawal symptoms might only occur when the interval between seizures is greater than the duration of seizure-induced enkephalin synthesis.(ABSTRACT TRUNCATED AT 400 WORDS)


Dynamic changes in chromaffin cell cytoskeleton as prelude to exocytosis. Earlier work by us as well as others has demonstrated that filamentous actin is mainly localized in the cortical surface of chromaffin cell. This F-actin network acts as a barrier to the chromaffin granules, impeding their contact with the plasma membrane. Chromaffin granules contain alpha-actinin, an anchorage protein that mediates F-actin association with these vesicles. Consequently, chromaffin granules crosslink and stabilize F-actin networks. Stimulation of chromaffin cell produces disassembly of F-actin and removal of the barrier. This interpretation is based on: (1) Cytochemical experiments with rhodamine-labeled phalloidin indicated that in resting chromaffin cells, the F-actin network is visualized as a strong cortical fluorescent ring; (2) Nicotinic receptor stimulation produced fragmentation of this fluorescent ring, leaving chromaffin cell cortical areas devoid of fluorescence; and (3) These changes are accompanied by a decrease in F-actin, a concomitant increase in G-actin, and a decrease in the F-actin associated with the chromaffin cell cytoskeleton (DNAse I assay). We also have demonstrated the presence in chromaffin cells of gelsolin and scinderin, two Ca(2+)-dependent actin filament-severing proteins, and suggested that chromaffin cell stimulation activates scinderin with the consequent disruption of F-actin networks. Scinderin, a protein recently isolated in our laboratory, is restricted to secretory cells and is present mainly in the cortical chromaffin cell cytoplasm. Scinderin, which is structurally different from gelsolin (different pIs, amino acid composition, peptide maps, and so on), decreases the viscosity of actin gels as a result of its F-actin-severing properties, as demonstrated by electron microscopy. Stimulation of chromaffin cells either by nicotine (10 microM) or high K+ (56 mM) produces a redistribution of subplasmalemmal scinderin and actin disassembly, which preceded exocytosis. The redistribution of scinderin and exocytosis is Ca(2+)-dependent and is not mediated by muscarinic receptors. Furthermore, our cytochemical experiments demonstrate that chromaffin cell stimulation produces a concomitant and similar redistribution of scinderin (fluorescein-labeled antibody) and F-actin (rhodamine phalloidin fluorescence), suggesting a functional interaction between these two proteins. Stimulation-induced redistribution of scinderin and F-actin disassembly would produce subplasmalemmal areas of decreased cytoplasmic viscosity and increased mobility for chromaffin granules. Exocytosis sites, evaluated by antidopamine-beta-hydroxylase (anti-D beta H) surface staining, are preferentially localized in plasma membrane areas devoid of F-actin.


Benzodiazepines in the brain. Their origin and possible biological roles. Great progress has been made in the last 5 yr in demonstrating the presence of benzodiazepines (BDZs) in mammalian tissues, in beginning studies on the origin of these natural compounds, and in elucidating their possible biological roles. Many unanswered questions remain regarding the sources and biosynthetic pathways responsible for the presence of BDZs in brain and their different physiological and/or biochemical actions. This essay will focus on recent findings supporting that: (1) BDZs are of natural origin; (2) mammalian brain contains BDZs in concentrations ranging between 5 x 10(-10)-10(-8) M; (3) dietary source of BDZs might be a plausible explanation for their occurrence in animal tissues, including man; (4) the formation of BDZ-like molecules in brain is a possibility, experimentally supported; (5) BDZ-like molecules including diazepam and N-desmethyldiazepam are elevated in hepatic encephalopathy; and (6) natural BDZs in the brain are involved in the modulation of memory processes. Future studies using the full range of biochemical, physiological, behavioral, and molecular biological techniques available to the neuroscientist will hopefully continue to yield exciting and new information concerning the biological roles that BDZs might play in the normal and pathological functioning of the brain.


Erythrocyte Ca, Na/K-ATPase in long-term streptozotocin diabetic rats. Effect of good glycemic control and a Ca antagonist. This study evaluated the effect of chronic hyperglycemia on erythrocyte membrane Ca and Na/K-ATPase activities in streptozotocin-induced diabetic rats. The activity of Ca-ATPase was significantly lower in diabetic than in normal rats. Good glycemic control by insulin restored the Ca-ATPase activity to normal. By contrast, diltiazem, a calcium entry blocker, had no effect on the enzyme activity. Calmodulin stimulated Ca-ATPase activity in all groups of rats. Na/K-ATPase activity was not altered in diabetic rats, and no effects of either insulin or diltiazem treatments were observed. The results suggest that erythrocyte Ca-ATPase activity is decreased in diabetic rats and is normalized by good glycemic control.


Reduction in cardiac conduction velocity delay by angiotensin converting enzyme inhibition in hypertensive patients with left ventricular hypertrophy. Detection by signal averaged electrocardiography. Hypertensive patients with left ventricular hypertrophy (LVH) have increased prevalence of ventricular arrhythmias. Slow conduction velocity at the level of hypertrophic myocardial cells has been one of the postulated mechanisms for these arrhythmias. To assess the effects of angiotensin converting enzyme inhibition on modification in ventricular conduction velocities, we studied 25 hypertensive patients with LVH using signal averaged electrocardiography (SAECG) in a randomized double-blind placebo controlled and cross-over trial. Data were acquired at baseline and 10 min after a double-blind intravenous infusion of saline placebo or 2.5 mg enalaprilat. Sequential cross-over was done the next day. Root mean square vector was 55 +/- 5 microV at baseline, 55 +/- 5 microV after placebo and 54 +/- 4 microV after enalaprilat (P = NS). Low amplitude signal < 40 msec was 45 +/- 4 msec at baseline, 45 +/- 4 msec after placebo, and 43 +/- 4 msec after enalaprilat (P = NS). There was no change in filtered QRS (fQRS) duration between baseline (113 +/- 10 msec) and placebo (113 +/- 11 msec) measurements. However, after enalaprilat infusion, there was a significant reduction in fQRS to 106 +/- 7 msec (P = .04), and five patients (20%) with late potentials had normalization of this feature (P = .001). The data suggest that angiotensin converting enzyme inhibition with enalaprilat reduces conduction velocity delay in hypertensive patients with LVH.


Reversal of cardiovascular structural changes when treating essential hypertension. The importance of the renin-angiotensin-aldosterone system. Our study attempted to evaluate the importance of changes in the circulating renin-angiotensin-aldosterone system (RAAS) and in hemodynamics in relation to observed changes in cardiovascular structure. We studied previously untreated men (n = 28) with essential nonmalignant hypertension and a supine casual diastolic blood pressure > 95 mm Hg on three to four separate (> 1-week interval) occasions measured in triplicate. We used intraarterial blood pressure, dye-dilution technique, plethysmography (hands), eye-ground photos, M-mode echocardiography, radio immunoassays, and multiple regression analysis. Patients were randomized to 6 months of double-blind treatment with either enalapril or hydrochlorothiazide, following 4 to 6 weeks on placebo. We found that enalapril blocked the plasma angiotensin converting enzyme (ACE) with a secondary increment in plasma renin activity (PRA) and reductions in angiotensin II (AII) and aldosterone. Blood pressure was lowered through a reduction in total peripheral resistance (TPR). Hydrochlorothiazide increased PRA, AII, and aldosterone, and lowered blood pressure mainly through a reduction in cardiac output. Enalapril was significantly more effective than hydrochlorothiazide in reversing structural changes in the retinal and hand vasculature as well as in the heart. A reduction in cardiac hypertrophy was seen even in the occasional enalapril-treated patient, in whom little or no reduction in blood pressure occurred. In the stepwise regression analyses, the changes in retinal and hand vascular structure were most strongly related to various changes in the RAAS, explaining 15 to 34% of the variance. For the changes in cardiac structure, the type of therapy (enalapril or hydrochlorothiazide) appeared to be the most important factor, explaining between 29 and 50% of the variance. The changes in cardiac structure were even more strongly related to changes in the RAAS for the enalapril treated patients and explained up to 55% of the variance in cardiac structure. It can be concluded that the reversal of structural vascular changes during antihypertensive therapy was more dependent on the blockade of the RAAS than on lowering of the blood pressure.


Excitable membranes, lipid messengers, and immediate-early genes. Alteration of signal transduction in neuromodulation and neurotrauma. The physical nature of neuronal cells, particularly in the functional and morphological segregation of synapse, soma, and dendrites, imparts special importance on the integrity of their cell membranes for the localization of function, generation of intrinsic second messengers, and plasticity required for adaptation and repair. The component phospholipids of neural membranes are important sources of bioactive mediators that participate in such diverse phenomena as memory formation and cellular damage following trauma. A common role for PAF in these processes is established through the suppressive effects of its antagonists. Furthermore, being both an extracellular and intracellular agonist of phospholipase activation, in addition to being a product of phospholipase activity, PAF assumes a centralized role in the cellular metabolism following neural stimulation. The linkage of PAF to neural immediate-early gene expression, both in vitro and in vivo, suggests that its effects are initiating to long-term formative and reparative processes. Such a common link between destructive and plastic responses provides an important view of cellular and tissue maintenance in the nervous system.


Species dependent effects of dihydroergosine on [3H]TBOB binding to membranes from the human, rat, bovine and mouse brain. Dihydroergosine enhanced [3H]TBOB binding to the crude synaptosomal membranes prepared from the whole rat brain and human frontal cortex. Higher concentrations of the same drug inhibited [3H]TBOB binding in the preparations obtained from the whole mouse brain and bovine frontal cortex. Bicuculline-induced enhancement and GABA- or diazepam-induced inhibition of [3H]TBOB binding were similar in the four species examined. The results indicate that dihydroergosine modulates species-dependently GABA/benzodiazepine receptors.


Paraneoplastic paralysis in a patient with malignant fibrous histiocytoma. A patient presented with lower limb paralysis and a large malignant fibrous histiocytoma (MFH) on her back. Metastatic disease to the spine was excluded and the diagnosis of paraneoplastic paralysis was made. This may be the first described case of a neuromyopathic paraneoplastic syndrome in malignant fibrous histiocytoma. Tissue culture and electron microscopy assisted in establishing the diagnosis of the tumour. A hitherto unrecognised endocrine factor may account for the hypokalaemia which was a feature in this patient.


High-dose chemotherapy supported with autologous bone marrow transplantation (ABMT) in germ cell tumors: a phase two study. In this paper, the first Italian multicentre experience with high-dose chemotherapy and ABMT in germ cell cancer is presented. Twenty-eight patients underwent treatment with a carboplatin-etoposide w/o ifosfamide high-dose combination. Seventeen patients were in progression of disease, 9 were responsive to salvage treatments or failed to achieve CR to front line, and 2 had stable disease (both with an elevated marker level) at the time of transplantation. Five patients, all of whom were in sensitive relapse at transplantation, are alive and disease-free at > 17 months' follow-up. Two patients died 15 days after ABMT, one of veno-occlusive disease and one of rapid uncontrolled tumor progression. In highly pretreated patients this schedule seems to provide an option of cure for a cohort of patients failing to achieve CR to standard salvage regimens for germ cell cancer. Definitive conclusions may eventually be drawn with a more homogeneous group of patients. This type of approach should continue to be taken in sensitive relapse patients only, as responses in progressive cases are very transient, with virtually no cures. The question of whether high-dose programs are better than standard chemotherapy will in any case be answered only in a randomized prospective trial.


Structure of bluetongue virus particles by cryoelectron microscopy. The structure of the bluetongue virus (BTV) particle, determined by cryoelectron microscopy and image analysis, reveals a well-ordered outer shell which differs markedly from other known Reoviridae. The inner shell is known to have an icosahedral structure with 260 triangular spikes of VP7 trimers arranged on a T = 13,l lattice. The outer shell is seen to consist of 120 globular regions (possibly VP5), which sit neatly on each of the six-membered rings of VP7 trimers. "Sail"-shaped spikes located above 180 of the VP7 trimers form 60 triskelion-type motifs which cover all but 20 of the VP7 trimers. These spikes are possibly the hemagglutinating protein VP2 which contains a virus neutralization epitope. Thus, VP2 and VP5 together form a continuous layer around the inner shell except for holes on the 5-fold axis.


Extensive small-cell lung cancer. A randomized comparison of two chemotherapy programs with early crossover in instances of failure. Association pour le Traitement des Tumeurs Intra-Thoraciques (ATTIT). Two chemotherapy regimens for patients with extensive small-cell lung cancer were prospectively compared in a randomized multicentric trial with crossover. Every four weeks, 60 consecutive previously untreated patients received either DPE (doxorubicin, cisplatin and etoposide), or CIV (carboplatin, ifosfamide and vincristine) with crossover as soon as progression or end of response were observed. Pretreatment characteristics were similar in the two groups. Fifty-seven patients were evaluated for response. The response rate was higher with the DPE regimen both in first-line (for DPE: response rate 62% (18/29), including 17% (5/29) of complete response (CR), and for CIV: response rate 29% (8/28) with no CR, p < 0.02) and in second-line after crossover (for DPE: response rate 45% (9/20) including 5% (1/20) of CR, and for CIV response rate 0%, p < 0.02). Major toxicities were equally frequent in both groups. No significant difference was found between median survival times (9.7 months for the DPE group and 10.4 months for the CIV group). We conclude that: 1) DPE is a more active regimen than CIV, both in first- and second-line; 2) no alternating scheme may be considered with these two combinations. Of particular note is the similar median survival times in the two groups, contrasting with the different activities of the two regimens.


Feasibility of quality of life assessment in a randomized phase III trial of small cell lung cancer--a lesson from the real world--the Swiss Group for Clinical Cancer Research SAKK. Between 1985 and 1990 the Swiss Group for Clinical Cancer Research (SAKK) for the first time assessed quality of life (QL) variables in 188 patients in a multicenter small-cell lung cancer trial that compared two different regimens of combination chemotherapy. QL-assessment was scheduled at the beginning of each of the six treatment cycles. The self-rating QL questionnaire included an early version of the EORTC QL questionnaire, a mood adjective checklist (Bf-S) and a single linear analogue scale (LASA) measuring general well-being. Compliance with completion of the scheduled questionnaires varied between 37% and 58% over the six cycles, and between 21% and 68% among the 7 participating institutions. Mean compliance was 49%. The institution was found the only significant factor predicting compliance (p < 0.001). Patient age, sex, education and biological prognostic factors at randomization were not predictors of compliance. Although compliance was poor, the data received was of high quality. We suggest practical guidelines for improving compliance with QL data collection in multicenter clinical trials.


A phase II study of high-dose epirubicin plus cisplatinum in advanced non-small-cell lung cancer (NSCLC). Thirty-seven patients with unresectable NSCLC received epirubicin (EPI) as i.v. bolus at the dose of 120 mg/sm+cisplatinum (CP) at the dose of 60 mg/sm every 28 days up to the maximum cumulative dose of 840 mg/sm of EPI. Of 35 evaluable patients, 19 (54%) (95% confidence limits: 37%-71%) achieved PR for a median duration of 10 months (range: 2-21). The majority of responsive patients experienced improvement in performance status, related-disease symptoms and body weight. Grades 3-4 leukopenia occurred in 42% of the patients. In five patients there was a > 10% reduction in the left ventricular ejection fraction as calculated by radionuclide angiocardiography. None of these patients suffered from cardiac symptoms. The median survival was 9 months (range 2-26). This study shows that inclusion of HD-EPI in a combination regimen contributes to obtaining a high remission rate in advanced NSCLC.


Detection of human papillomavirus DNA in seborrheic keratosis by polymerase chain reaction. We searched for HPV DNA in 72 cases of seborrheic keratosis (nongenital SK 29, genital SK 43) using polymerase chain reaction. Amplification of HPV DNA was found in 24 (nongenital, 1; genital, 23). HPV DNA type 6 was present in 18 cases and type 11 in four and type 33 in two. Twenty-three (53%) cases of genital region SK and one (3%) case of nongenital region SK contained HPV DNA sequences. Histopathologically, the perinuclear vacuolization of epithelial cells, mitosis and dyskeratotic cells were more marked in HPV DNA-positive SK than those in negative SK (P < 0.01). We suppose HPV may play an important role in epidermal proliferation of SK in the genital region.


Interaction of beta-carboline with chick embryo spontaneous motility. The effect of beta-carboline (beta-CCE) on spontaneous motility and its development was studied in chick embryos between the 11th and 19th day of incubation. 1. Acutely administered beta-CCE (7.5 mg/kg e.w.) already induced significant activation of motility in 11-day-old embryos. From the 17th day of incubation activation acquired a paroxysmal character. 2. In spinal embryos (decapitated on the second day of incubation) there was no such activating effect, demonstrating that it is associated with supraspinal components of the CNS. 3. In chronic administration from the fourth day of incubation (1.55 +/- 0.24 mg/kg e.w./24h), beta-CCE led to reduced development of spontaneous motility. The effect was concentrated in the period between the fourth and eighth day of incubation. The chronic administration of beta-CCE augmented the activating effect of metrazol and weakened GABA-inhibition of spontaneous motility. 4. On the basis of their findings, the authors express the hypothesis that the benzodiazepine beta-CCE-sensitive component of the complex GABA receptor evidently already functions from the beginning of the second half of incubation of chick embryos.


The increase in cAMP and cGMP levels in the oestrogenized rat hypophysis. Male and female rats were given oestradiol benzoate (1 mg s.c. twice a week for 3 weeks) and/or sodium nitroprusside (SN), a donor of nitric oxide (NO), which was administered in their food in amounts of 0.2 or 0.6 mg/rat/day. Neither oestradiol-induced hypertrophy of the hypophysis, nor the serum prolactin (PRL) level, was affected by the simultaneous administration of SN. The PRL content of the hypophysis rose after oestradiol in the males, but the increase was again uninfluenced by the simultaneous administration of SN and the cAMP content of the hypophysis--raised after oestradiol--was likewise unaffected. The amount of cGMP in the hypophysis after oestradiol rose only in males. Both the serum and the hypophyseal prolactin level were found to be correlated to the cAMP and the cGMP content of the hypophysis. It was found that the simultaneous administration of SN together with oestradiol slightly reduced the increase in the cGMP content of the hypophysis elicited with oestradiol treatment only.


Opioid receptors in the rat spinal cord after longlasting deafferentation. In vitro binding of specific opioid ligands to their respective sites in membrane fractions and the contribution of individual receptor classes (mu, delta, kappa) was studied in rats after longlasting (up to 22 months) section of spinal dorsal roots at the cervical (C5-8) or thoracic (Th1-4) level. This procedure leads to autotomy or scratching of the skin on the operated side. The total number of receptors in the cervical and thoracic spinal cord was more than doubled in both operated and contralateral part of the cord in comparison with intact controls of the same age. In the cervical region, this increase mainly represented a rise in the number of free receptors, whilst in the thoracic region both free and saturated receptors were increased. On the deafferented side, receptor selectivity, especially in the delta and kappa types was decreased.


Complexoproductive and antiheparin properties of low density lipoproteins (LDL). I. Interaction of isolated low density lipoproteins with heparin of different molecular weight. Low density lipoproteins/LDL/ form unsoluble complexes with heparin of different molecular weight in the presence of CaCl2. Quantity of the complexes depends on molecular weight of heparin, concentration of reagents and pH of the reacting medium. The biggest quantity of complexes result from LDL-heparin of the molecular weight 16000, the quantity is smaller with heparin of the molecular weight 5100 and the quantity is smallest with heparin of the molecular weight 3700. Components of LDL--heparin complexes are bound by means of ion and hydrogen bonds.


Complexoproductive and antiheparin properties of low density lipoproteins (LDL). II. Interaction of serum low density lipoproteins (LDL) with heparin of different molecular weight. Quantity of unsoluble LDL complexes with heparin of different molecular weight in hypo-, normo-, and hyperlipemic blood serum in the presence of CaCl2 was estimated. At the same weight concentration the biggest quantity of LDL complexes are formed with heparin with heparin of the molecular weight 16000, the quantity is smaller with heparin of the molecular weight 5100, and it is the smallest with heparin of molecular weight 3700.


The effect of collagen degradation products (CDP) on the central nervous system (CNS). Rat collagen type I was digested with bacterial collagenase. The peptides obtained were submitted to gel filtration on Sephadex G-25. Two fractions differing in molecular weight (CDP I-3000 D and CDP II-1200 D) were obtained. These fractions administered to rats were found to have an inhibiting effect on the central nervous system (CNS) in Lat's test and the mobility of the animals recorded on a motimeter was found to be reduced. This effect was dependent on the peptide dose and occurred after intravenous (iv) as well as intraventricular (icv) injection.


Cellular signaling in thymocyte apoptosis. Induction of apoptosis (programmed cell death) in response to T cell receptor triggering is now thought to be involved in the process of negative selection in the thymus, and current work is therefore aimed at investigating how apoptosis is regulated within the cells. To this end, recent work has implicated several of the well-known signal transduction pathways already known to regulate T cell activation in the regulation of apoptosis in thymocytes. In particular, elevations of the cytosolic Ca2+ level or increases in cAMP can trigger thymocyte apoptosis, whereas activation of protein kinase C appears to inhibit apoptosis in response to either induction pathway. Moreover, crosslinking of Thy-1, CD4, or CD8 leads to potentiation of T cell receptor-mediated cell death, effects that appear to involve protein tyrosine kinase activation. These observations may be relevant to the question of how T cell receptor occupancy can mediate both differentiation and death during intrathymic T cell development.


The absence of genital human papillomavirus DNA in virginal women. Our aim was to determine the prevalence of human papillomavirus (HPV) infection in virginal women and to establish if other forms of sexual activity not involving penetration may contribute to the transmission of this infection. Female patients attending medical practitioners, and female high school students were recruited. Each participant answered an anonymous self answer questionnaire and collected a tampon specimen. Tampons were analysed for the presence of human papillomavirus by the polymerase chain reaction (L1 general primer and probe). Fifty-five female participants who were of a mean age of 18 years (range 13-41) were recruited. Twenty-three reported non-penetrative sexual activity. None of the 55 women had evidence of HPV infection (0%, upper 95% confidence interval 5.4%). Twelve tampons from sexually active women with normal Papanicolaou smears and a mean age of 26 years (range 19-44) attending a hospital clinic were analysed at the same time and four were found to be positive (P = 0.0006). We conclude that HPV infection prior to sexual intercourse, as determined by tampons specimens, is rare, even in those participating in other forms of sexual activity.


The apoptosis endonuclease and its regulation. Activation of an endogenous endonuclease has been observed in conjunction with the structural changes of apoptosis in a wide variety of cell types and circumstances. The endonuclease is present constitutively in some cells (e.g. rodent cortical thymocytes) in which apoptosis is readily triggered by many unrelated stimuli, but is inducible in others. Purification of this enzyme is an objective of some importance in apoptosis research, as it might act as a marker of susceptibility to apoptosis and lead to better understanding of the regulation of the process as a whole. Early data suggest that the thymocyte endonuclease is an anionic protein of molecular weight greater than 110 kDa, with a pH optimum of 7.5 and a double-strand cleavage preference. Its activity, and the induction of apoptosis as a whole, is regulated by several familiar cellular proto-oncogenes and oncosuppressor genes, including c-myc, Ha-ras, bcl-2 and p53.


The purification of nucleoside diphosphate kinase from rat brain. Nucleoside diphosphate kinase from the cytosol of rat brain was purified to electrophoretic homogeneity through a series of chromatography columns, including DE-52, AcA-34 Ultrogel, phenyl-Sepharose and Q-Sepharose. The molecular weight of the enzyme was 64,000 as estimated by gel filtration, and it consisted of 3 identical 19,000 subunits. [35S]-GTP gamma S binding and autophosphorylation of the enzyme, and inhibition of the GTPase activity of G-protein were used to screen for the enzyme. These methods were shown to be useful for purifying and assaying the enzyme.


Detection of circulating antigen by McAb-AST for evaluating the efficacy of anti-Leishmania chemotherapy. We have adapted the simple and sensitive McAb-antigen spot test (AST) for evaluating the efficacy of anti-Leishmania chemotherapy. Serum samples from 37 kala-azar patients were tested by McAb-AST, and all showed definite positive reactions before treatment. After a course of antimony treatment, 20 turned negative, coupled with the disappearance of clinical symptoms; another 12 cases responded with weak positivity accompanied by an improvement of clinical manifestations; and the remaining 5 antimony-resistant patients showed strong positive reactions, with their conditions gradually worsening. Furthermore, all 6 cases in which the diagnosis was missed by the bone marrow smear method turned McAb-AST negative after chemotherapy. These results suggest that McAb-AST can be used to evaluate the efficacy of chemotherapy as well as to avoid missed diagnosis by the bone marrow smear method.