Analysis of occlusal stresses transmitted to the inferior alveolar nerve by multiple threaded implants.

BACKGROUND: Potential nerve injury or loss of sensation can occur after mandibular implant placement or loading. To avoid this type of damage, it is critical to determine the proper distance from implants to the mandibular nerve. Hence, the purpose of this study is to use biomechanical analyses to determine the safe distance from multiple implants to the inferior alveolar nerve. METHODS: Using the boundary element method, a numerical mandibular model was designed to simulate a mandibular segment containing multiple threaded fixtures. This model allows assessment of the pressure, as induced by occlusal loads, on the trigeminal nerve. Such pressure distribution was evaluated against different distances from the fixtures to the mandibular canal, against the possible lack of the central fixture in a three-abutment configuration, and against different levels of implant osseointegration. All the simulations considered a canal that is orthogonal to the implant axis. RESULTS: Nerve pressure increased quickly when the implant-canal distance decreased in the range studied. Lack of the central implant to support the central abutment caused major increases in nerve pressure. CONCLUSIONS: This study suggests a minimal implant-canal distance of 1 mm to prevent inferior alveolar nerve damage caused by three connected implants. For clinical safety, an additional 0.5 mm is recommended as a cushion, so a 1.5-mm minimal distance should be planned to avoid potential nerve injury.

Analysis of the occlusal stress transmitted to the inferior alveolar nerve by an osseointegrated threaded fixture.

BACKGROUND: Altered sensation can occur after the placement or loading of mandibular implants. Limited evidence exists with regard to the proper distance between the implant and the mandibular nerve to ensure the nerve's integrity and physiologic activity. The proper distance should come from evaluation of clinical data as well as from biomechanical analyses. METHODS: A numeric mandibular model based on the boundary element method was created to simulate a mandibular segment containing a threaded fixture so that the pressure on the trigeminal nerve, as induced by the occlusal loads, could be assessed. Such pressure distributions were evaluated with different distances of the fixture from the mandibular canal and considering different bone densities. Although all simulations considered a canal that was orthogonal to the implant axis, in one case, the effects of an inclined canal were analyzed. RESULTS: The nerve pressure increased rapidly with a bone density decrease. A low mandibular cortical bone density caused a major nerve pressure increase. CONCLUSION: Our study suggested a distance of 1.5 mm to prevent implant damage to the underlying inferior alveolar nerve when biomechanical loading was taken into consideration.

Monovalent modified-live vaccine against bluetongue virus serotype 2: immunity studies in cows.

A challenge study was conducted to determine the efficacy of vaccination against bluetongue (BT) virus (BTV) serotype 2 in protecting cattle against infection. A group of 30 cows, vaccinated seven months previously with monovalent BTV-2 modified-live vaccine produced by Onderstepoort Biological Products in South Africa, were challenged subcutaneously with 2x 10(5.8)TCID50/ml of BTV-2 field isolate. All cattle originated from the same population in the Sardinian province of Oristano. Eight unvaccinated calves from a BTV-free herd also participated in this study; four were inoculated with BTV-2 and used as positive controls whilst the remaining four were used as negative controls to confirm that no BTV was circulating locally. Blood samples were taken from all animals three times a week for two months. Serum samples were tested for antibody against BTV using the competitive enzyme-linked immunosorbent assay (c-ELISA) and the virus neutralisation (VN) test. Virus isolation was attempted on the blood samples by intravenous egg inoculation followed by two blind passages in Vero cells. Virus titres following challenge were determined also. Of the 30 cows vaccinated, 29 were positive in the c-ELISA and demonstrated neutralising antibodies. At the time of challenge, 11 cows had no virus neutralising antibody while the remainder had low titres ranging from 1:10 (11 cows) to 1:20 (6 cows); two cows showed titres of 1:40 and 1:80, respectively. None of the cows showed signs of disease after challenge and no BTV was isolated from the blood of the 29 cows that had developed antibodies after vaccination. Commencing on day 9 post challenge, BTV-2 was isolated from the blood of the single cow that had not seroconverted following vaccination and from the blood of the unvaccinated controls. Viraemia lasted until day 21 post challenge. Neither BTV nor antibody was detected in the blood samples taken from the negative control group. These observations indicate that the monovalent BTV-2 modified-live vaccine protects most animals when challenged with field virus seven months post vaccination.

Virological and serological response of cattle following field vaccination with bivalent modified-live vaccine against bluetongue virus serotypes 2 and 9.

Following the bluetongue (BT) epidemic in Italy, the government initiated a vaccination campaign involving all domestic ruminants (cattle, sheep and goats) in the affected and adjacent areas to create a resistant population and to reduce virus circulation. Based on the serotypes circulating in the affected areas, monovalent BT virus (BTV) serotype 2 (BTV-2), or bivalent BTV-2 and BTV-9, modified-live vaccines were used. These are manufactured by Onderstepoort Biological Products in South Africa and, because they are recommended for use in sheep only, very little data exists on their use in cattle under field conditions. To evaluate duration and levels of viraemia and the antibody response following vaccination, 30 cattle in various stages of pregnancy were selected and vaccinated with a bivalent BTV-2/BTV-9 vaccine. Blood samples were taken from the animals three times a week for two months and screened using the competitive enzyme-linked immunosorbent assay (c-ELISA) and the virus neutralisation (VN) test. Intravenous egg inoculation, followed by two blind passages in Vero cells, was used to isolate BTV-2 and BTV-9 from ethylene-diaminetetra-acetic acid (EDTA) blood samples, and virus titres determined in viraemic animals. Titres against BTV were detected in 27 animals between days 4 and 35 post vaccination (pv). Viraemia peaked on day 9 pv with average viral titres of 10(4.5)TCID50/ml. From day 9 pv, the c-ELISA detected antibodies in all animals while low VN titres were observed commencing on day 18 pv. Furthermore, VN antibody to BTV-2 was detected in only 17 of the animals vaccinated and to BTV-9 in 27 animals.

Novel multidrug resistance reversal agents.

A series of 59 alpha-aryl-alpha-thioether-alkyl, -alkanenitrile, and -alkanecarboxylic acid methyl ester tetrahydroisoquinoline and isoindoline derivatives (15a-48) were synthesized and evaluated as multidrug resistance (MDR) reversal agents. The compounds were tested on S1-B1-20 human colon carcinoma cells selected for resistance to bisantrene. Both the cytotoxicity of the reversal agents and their ability to resensitize the cells to bisantrene were determined. All but two of these compounds (15q, 40) were more effective MDR reversal agents in vitro than verapamil (VRP), a calcium channel antagonist which also has been shown to possess MDR modulating activity. Several showed good activity in this assay (IC50's < 0.5 microM), the most potent being isoindolines 44 (IC50 0.26 microM) and 46 (IC50 0.26 microM) and tetrahydroisoquinolines 47 (IC50 0.29 microM) and 15m (IC50 0.30 microM). A number of compounds were evaluated in vivo against vincristine (VCR)-resistant murine P388 leukemia, as well as against human epidermoid carcinoma KB/8.5 implanted sc in athymic mice. The reversal agents which consistently showed the highest activity, together with low toxicity, were alpha-aryl-alpha-thiotolylalkanenitrile tetrahydroisoquinoline derivatives with electron-rich alkoxy substituents on the aromatic rings. Of the tested compounds, the most effective reversal agents for both tumor lines were 15h (33% increased life span at 12.5 mg/kg, 0.2 mg/kg VCR versus VCR alone in the VCR-resistant P388 leukemia model and 59% relative tumor growth at 50 mg/kg, 8 mg/kg doxorubicin versus doxorubicin alone in the KB/8.5 model) and 39a (48% increased life span at 50 mg/kg, 0.2 mg/kg VCR versus VCR alone in the VCR-resistant P388 leukemia model and 46% relative tumor growth at 25 mg/kg, 8 mg/kg doxorubicin versus doxorubicin alone in the KB/8.5 model). The mechanism of action of these compounds is believed to involve blocking the drug efflux pump, P-glycoprotein.

Tumor inhibitory activity of anti-ras ribozymes delivered by retroviral gene transfer.

This study reports the successful introduction into tumor cells of a ribozyme directed against an oncogene using a retroviral gene delivery system. A hammerhead ribozyme that selectively targets and cleaves the activated Ha-ras (V12Ras) oncogene was delivered using a retroviral vector and expressed under the control of a transfer RNA promoter in V12Ras-transformed 3T3 murine fibroblast and rat colon epithelial cells. The expression of V12Ras messenger RNA and V12Ras P21 protein was reduced by up to 100% and 75%, respectively, in cells transduced with functional ribozyme. Reductions in V12Ras expression correlated with the stable expression of the functional ribozyme in vivo and decreased tumorigenicity in nude mice. Ribozyme constructs containing identical antisense flanking regions, but a mutant catalytic center, did not attenuate V12Ras expression or decrease the tumorigenicity of transduced tumor cells. These data support the potential therapeutic role of antioncogene ribozymes.

N-phosphoryl derivatives of bisantrene. Antitumor prodrugs with enhanced solubility and reduced potential for toxicity.

The selective phosphorylation of bisantrene (1) affords bis(phosphonoguanidinic acid) 6, a prodrug with enhanced aqueous solubility (as sodium salt 7) at physiological pH. Unlike 1, in a rat tail vein model, no precipitation was observed when bis(phosphonoguanidinic acid) 6 was injected. While in rats 6 hydrolyzed to monophosphonoguanidinic acid 9 with a half-life of ca. 12 min., complete hydrolysis to bisantrene required several hours. The corresponding monophosphonoguanidinic acid 9 was synthesized from bisantrene and also showed good solubility and antitumor activity. While the antitumor activities of 6 in mice were comparable to bisantrene against B-16 melanoma and P-388 and L-1210 leukemias, it was inactive in vitro vs several tumor cell types. Thus, its activity in vivo resulted from its ability to serve as a prodrug for bisantrene.

A new family of water-soluble, third generation antitumor platinum complexes.

[1,1-Cyclobutanedicarboxylato(2)-O,O'](1,3-dioxane-5,5-dimethan amine- N,N')platinum(II), 3a, a third generation, very water-soluble platinum complex, has been synthesized along with several of its analogues. All members of the new family contain a 1,3-dioxane or 1,3-dioxolane-1,3-diamine as their basic ligand, a moiety which contributes to their increased water solubility, and a bidentate acid ligand, which is responsible for their good stability. They were all easily crystallized and characterized by 1H NMR and elemental analysis, and the parent complex 3a was further characterized by 13C NMR. Their very desirable physical properties combined with their broad spectrum of antitumor activity and reduced toxicity make them good candidates of further development.

Water-soluble third generation antitumor platinum complexes, [2,2-bis (aminomethyl)-1,3-propanediol-N,N']-[1,1-cyclobutanedicarboxylato (2-)-O,O']platinum(II) and [1,1-cyclobutanedicarboxylato(2-)-O,O'] [tetrahydro-4H-pyran-4,4-dimethanamine-N,N']platinum(II).

The synthesis, stability, and antitumor activity of a series of water-soluble third generation platinum(II) complexes have been described. Among these complexes, [2,2-bis(aminomethyl)-1,3- propanediol-N,N'] [1,1-cyclobutanedicarboxylato(2-)-O,O']platinum(II) and [1,1-cyclobutanedicarboxylate(2-)-O,O'](tetrahydro-4H-pyran-4,4- dimethanamine-N,N'-)platinum(II) have shown the greatest promise for further investigation and are currently under clinical evaluation.

Tissue deposition of bisantrene in B16 melanoma tumor-bearing mice.

Single doses of 14C-labeled bisantrene, a new antitumor agent, were administered intravenously at 10 and 100 mg/kg to mice bearing intraperitoneally implanted B16 melanoma. At 24 hr after dosing, the tumors contained relatively high drug concentrations as compared to most of the other tissues. The concentrations averaged 2.4 and 28.3 micrograms/g tumor and the tumor/blood concentration ratios were 40/1 and 29/1 for the low and high doses, respectively.

Modulation of the immune response to tumors by a novel synthetic compound, N-[4-[(4-fluorophenyl)sulfonyl]phenyl] acetamide (CL 259,763).

CL 259,763, N-[4-[(4-fluorophenyl)sulfonyl]phenyl] acetamide, is an orally active compound capable of modifying the reactivity of certain lymphoid cell populations affected by the growth of a tumor. The compound augmented the response of lymphocytes from tumor-primed animals to syngeneic tumor cells, resulting in a marked increase in tumor cell destruction. Likewise, it enhanced macrophage inhibitory effects on the growth of tumor cells in vitro. These "activated" macrophages were detectable in peritoneal exudates of treated mice 4 to 12 days after receiving a single oral dose of CL 259,763, with peak activity being demonstrable by day 7. The compound also restored the alloreactivity of lymphocytes from immunodepressed mice bearing the Lieberman plasma cell tumor, possibly by interfering with suppressor cells. Macrophages and lymphocytes from treated mice released significantly more IL-1 and IL-2-like factors in culture than did the control counterparts. Sera from treated mice also possessed more colony stimulating factor than those from normal mice. Immunoadjuvant effects were evident when the compound was administered with an inactivated L1210 leukemia vaccine and it enhanced the effectiveness of cytotoxic chemotherapy when given to mice challenged with P388 murine leukemia. These immunomodulating effects of CL 259,763 may hopefully be exploited in efforts to augment the immune response of the host to a progressively growing tumor.

Molecular and biochemical pharmacology of mitoxantrone.

Evidence has been presented which indicates that Nv: intercalates DNA and additionally causes inter- and intra-strand crosslinking possibly associated with its charged side arms; there is an apparent preference for G-C base pairs; induces single strand and double strand breaks in DNA; strongly inhibits DNA and RNA synthesis; causes nuclear aberrations and chromosomal scattering; induces a block in the G2 phase of the cell cycle with an increase in cellular RNA and polyploidy; is not cell cycle phase-specific with respect to cell kill; does not induce free-radical formation; does not induce lipid peroxidation or superoxide formation; rather it may inhibit ADR-stimulated lipid peroxidation and microsomal superoxide production; does not appear to have a strong potential for cardiotoxicity on the basis of currently postulated mechanisms of action; is capable of inducing cellular resistance in vitro; resistance is associated with an apparent alteration in the cell membrane impairing drug transport into the cell. Although the precise mechanism(s) of tumor cell killing has not been fully defined it is most likely associated with an interaction by Nv with chromosomes resulting in DNA damage, which if not efficiently repaired, will lead to inhibition of nucleic acid synthesis and eventual cell death.

Synthesis of 3,6-bis(aminoalkoxy)acridines and their effect on the immune System.

A series of 3,6-bis(aminoalkoxy)acridines (2) was prepared and shown to have a protective antiviral effect against an interferon-sensitive virus (Columbia SK) and to partially restore an antibody response to a T-cell-dependent antigen in leukemic immunosuppressed mice. The presence of circulating interferon and the stimulation of natural killer cell activity in mice was observed for 21.

Antitumor agents. 2. Bisguanylhydrazones of anthracene-9,10-dicarboxaldehydes.

9,10-Anthracenedicarboxaldehyde bis[(4,5-dihydro-1H-imidazol-2-yl)hydrazone] (bisantrene, VI-1) showed anticancer activity in mice vs. both leukemias and solid tumors. Increases in life span vs. the following neoplasms were: P-388 leukemia, 137%; B-16 melanoma, 122%; Lieberman plasma cell tumor, greater than 85%; colon tumor 26, 150%; Ridgway osteogenic sarcoma, 85%. There were significant numbers of long-term survivors. Both DNA and RNA synthesis were strongly inhibited. The drug was resistant to biodegradation and was bound strongly to tissues; in monkeys the half-life for disappearance from serum was 6 days. Related hydrazones were synthesized, and structure-activity relationships are discussed. Two routes to ring-substituted anthracene-9,10-dicarboxaldehyde intermediates were developed.

Activity of a novel anthracenyl bishydrazone, 9,10-anthracenedicarboxyaldehyde Bis[(4,5-dihydro-1H-imidazol-2-yl)hydrazone] dihydrochloride, against experimental tumors in mice.

9,10-Anthracenedicarboxaldehyde bis[(4,5-dihydro-1H-imidazol-2-yl)hydrazone] dihydrochloride (CL 216942; bisantrene hydrochloride; NSC 337766), a member of a new chemical class of compounds with antineoplastic properties, has been evaluated for antitumor activity in experimental murine tumor systems. The compound produced significant increases in life span (LS) and long-term survivors among mice bearing transplantable leukemias and solid tumors. Optimal treatment regimens resulted in an ILS of greater than 173 and 151% in mice with P388 and L1210 leukemia, respectively, an ILS of greater than 85% in mice with Lieberman plasma cell tumor, and an ILS of greater than 200, 150, and 63%, respectively, in mice with B16 melanoma, colon tumor 26, and Ridgway osteogenic sarcoma. An adriamycin-resistant subline of P388 leukemia showed complete cross-resistance to CL of 216942. The compound was active when administered by the i.p., i.v., and s.c. routes, but p.o. activity was not observed. Significant schedule dependency was not observed when the drug was administered once daily for 9 days, once every 4 days, or as a single dose, but single doses typically produced the best effects. CL 216942 was a potent inhibitor of DNA and RNA synthesis in L5178Y lymphoma cells cultured in vitro, and preliminary studies indicated the drug was a DNA-intercalating agent. The drug was cytotoxic for rapidly proliferating and nonproliferating (G0) human colon carcinoma WiDR cells in vitro.U

Antitumor agents. 1. 1,4-Bis[(aminoalkyl)amino]-9,10-anthracenediones.

The condensation of alkylenediamines with quinizarin or with 2,3-dihydro-1,4,5,8-tetrahydroxy-9,10-anthracenedione, followed by oxidation, gave 1,4-bis[aminoalkyl)amino]-9,10-anthracenediones. Some of these compounds and their 2,3-dihydro derivatives were markedly active against both leukemias and solid tumors in mice. Activity was maximal with 5,8-dihydroxylation and 1,4-bis[(2-aminoethyl)amino] substitution, in which the terminal nitrogen atoms were either unsubstituted (compound 50) or carried 2-hydroxyethyl groups (compound 40), indicating the importance of hydrophilicity. Against B-16 melanoma, 50 gave greater than 433% increase in median life span (ILS) with 7/10 80-day survivors. Against P-388 leukemia, 40 gave greater than 500% ILS with 4/5.60-day survivors; its efficacy and therapeutic index equaled or surpassed those of adriamycin, cyclophosphamide, daunorubicin, methotrexate, or 5-fluorouracil. Against L-1210 leukemia, B-16 melanoma, and colon tumor 26, 40 was generally as effective or more effective than adriamycin and is now undergoing preclinical toxicological evaluation.

Studies on vaccinia virus-directed deoxyribonucleic acid polymerase.

A vaccinia-directed deoxyribonucleic acid (DNA) polymerase has been partially purified from the cytoplasmic fractions of virus-infected HeLa cells. The utilization of natural and synthetic templates by this enzyme resembles that of the host cell DNA-dependent DNA polymerases. The vaccinia DNA polymerase cannot copy ribopolymers or ribonucleic acid but is very effective with an "activated" DNA as template. An exonuclease preferring single-stranded DNA as substrate is found in the most highly purified preparations of the enzyme. The molecular weight of the vaccinia DNA polymerase seems to be about 110,000. The viral DNA polymerase is also found to be associated with purified, infected cell nuclei, and this association may be due, at least in part, to nonspecific adsorption of the vaccinia DNA polymerase by nuclei.

Polyphasic taxonomy of the genus Vibrio: polynucleotide sequence relationships among selected Vibrio species.

Polynucleotide relationships among selected Vibrio species were examined by means of deoxyribonucleic acid (DNA) reassociation reactions and chromatography on hydroxyapatite. Relative levels of intraspecific DNA duplex formation (V. cholerae-V. cholerae and V. parahaemolyticus-V. parahaemolyticus) were found to be high at 60 C (>80%), and only minimally reduced at 75 C. Interspecific DNA duplexes between V. cholerae DNA and that of the non-cholera vibrios also exhibited high relative levels of formation at 60 C (>80%) and, with one exception, were only slightly reduced at 75 C. The thermal stability of these duplexes formed at 60 or 75 C was virtually identical to that of homologous V. cholerae DNA duplexes. The degree of reassociation and the thermal stability of V. cholerae-non-cholera vibrio DNA duplexes suggests relatively little evolutionary divergence in these organisms. In all other interspecific DNA reassociation reactions, only low levels of DNA duplex formation were noted at 60 C (<25%), and these were drastically reduced (>50%) at 75 C. The degree of nucleotide sequence divergence indicated by these reactions suggests that these Vibrio species are not significantly related to V. cholerae or V. parahaemolyticus. Reassociation reactions between V. cholerae DNA and the DNA of V. parahaemolyticus indicated these species were not significantly related to each other.