Mutants from V79 fibroblasts exhibiting hypersensitivity to aphidicolin and 3′-azido-3′-deoxythymidine

One variant, aph^hs-3 was previously isolated based on a hypersensitivity to nontoxic concentrations of aphidicolin, a specific inhibitor of DNA polymerases-alpha and delta. This variant was found to be more sensitive to temperatures above 35°C and to 10 µM of 3-azido-3-deoxythymidine (zidovudine, azidothymidine, or AZT) than the parental 743x cells. DNA polymerase activities in the cell extract or in the partially purified fraction by DEAE-cellulose (DE52) anion exchange column from aph^hs-3 were active at 40°C. No significant differences in deoxynucleoside triphosphate pools were observed at 34°C for both the parental 743x and aph^hs-3 cells. Revertants were isolated at 39°C: six revertants (aph^hs-3-tr1 through aph^hs-3-tr6) were obtained without aphidicolin; one revertant aph^hs-3-tar (the tar clone) was selected in aphidicolin (0.12 µM). The hypersensitivity to aphidicolin (Aph^hs) and AZT (AZT^hs) was cosegregated in the revertant aph^hs-3-tr5 (the tr5 clone), while the tar clone was not AZT^hs. There was a similar increase in the specific activity of³H-labeled DNA in all cell lines after additions of [³H]AZT or [³H]thymidine. Additions of purine or pyrimidine arabinosides (araT, araC, and araA) to all cell lines resulted in a similar cytotoxicity, suggesting the anabolism of dTTP was not defective in the tr5 clone. The spontaneous mutation rate at the hypoxanthine-guanine phosphoryltransferase locus using replating techniques and 6-thioguanine resistance selection was 5×10^–7, 2.2×10^–6, or 1.3×10^–6 per generation for the tr5, 743x, or tar cell lines, respectively. Most importantly, DNA polymerase activities in the cell extract of the revertant tr5 clone were inhibited by 0.5 µM AZTTP. In contrast, no inhibition was observed in those of the parental 743x and revertant tar cells. The cosegregation of both Aph^hs and AZT^hs in the tr5 revertant suggests that these two phenotypes may be a result of the same mutational event.     

Alkaline phosphatase and peptidase levels in invertebrate cartilage

Summary Cartilage is encountered in the skeletons of many advanced invertebrates, yet it never calcifies or is replaced by bone. In an attempt to account for the absence of bone in invertebrates, we tested a hypothesis proposing that absence or inadequate quantities of several enzymes associated with vertebrate osteogenesis may underlie the failure of the invertebrates to evolve bone. The enzymes examined were alkaline phosphatase, alanyl -naphthylamidase, and neutral protease. Their activities were measured in the gill cartilage of the Atlantic horseshoe crab, Limulus polyphemus, and the odontophore cartilage of the marine whelk, Busycon canaliculatum. Animals were collected from the Cape Cod area. Samples of cartilage of Limulus perichondrium, various nonskeletal tissues, and neonatal rat calvaria, the latter as a reference standard, were homogenized in 0.1 M phosphate buffer (pH 7.1) and analyzed for protein content and the above-mentioned enzyme activities. Alkaline phosphatase specific activity was readily detected in most tissues except the invertebrate cartilage specimens in which it was present only at near-trace levels. Naphthylamidase and protease activities were present in all tissues. In a single experiment, higher phosphatase values were recorded for Limulus cartilage retaining perichondrium, but in a subsequent trial assaying cartilage retaining perichondrium, denuded cartilage, and isolated perichondrium separately, it was demonstrated that phosphatase activity resided primarily within the perichondrium. Exposure of thick cryostat sections to p-nitrophenyl phosphate confirmed the suspicion that alkaline phosphatase activity was present principally in the perichondrium. In view of the strong association between alkaline phosphatase and vertebrate cartilage mineralization it is proposed that extremely low levels of this enzyme may contribute to the failure of invertebrate cartilage to be replaced by bone.     

Hepatic abscess

Hepatic abscess—amebic or pyogenic—can be diagnosed with great accuracy by either ultrasonography or computed tomographic (CT) scanning. Ultrasound is the modality of choice and will detect almost 100% of abscesses. Confirmation of a diagnosis of amebic liver abscess is made by the indirect hemagglutination test that should be positive in almost 100% of cases. Cultures of pus from the abscess and from the blood must be obtained in cases of pyogenic liver abscess. A positive culture of pus from the abscess has been achieved in 90% of cases. Ultrasound or CT guidance is utilized in aspiration of a hepatic abscess. In the treatment of an amebic liver abscess, metronidazole is the amebicide of choice. Open drainage is contraindicated. For cases that fail to respond to therapy with amebicides, closed drainage guided by CT or ultrasound is performed. Secondary bacterial infection of an amebic liver abscess is an extremely rare event. The identification and determination of the antibiotic sensitivity of organisms responsible for pyogenic liver abscess is a crucially important step. Unless a celiotomy is necessary to correct an intraabdominal process or the abscess is extremely large, the initial treatment of pyogenic liver abscess is a 2 week course of appropriate antibiotics followed by a 1 month course of oral antibiotics. The majority of pyogenic liver abscesses will respond to such treatment. If drainage of a pyogenic abscess is required, the preferable technique is with a percutaneous CT- or ultrasound-directed catheter. Open surgical drainage should be reserved for those cases in which a celiotomy is required for other purposes or for the patient who has failed a course of appropriate antibiotic therapy and closed percutaneous drainage is not feasible. The mortality for treatment of amebic liver abscess should be approximately zero and for pyogenic liver abscess should be less than 10%.

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Resumen El absceso hepático—amibiano o piogénico—puede ser diagnositicado con gran precisión mediante la ultrasonografía (US) o la tomografía computadorizada (TC). La ultrasonografía es la modalidad de escogencia; détecta casí el 100% de los abscesos. La confirmación del diagnóstico de absceso amibiano del hígado se hace por la prueba de hemaglutinación indirecta, la cual debe resultar positiva en prácticamente el 100% de los casos. Cultivos del pus y de la sangre deben ser realizados en los pacientes con abscesos piógenos. Se logran cultivos positivos del pus del absceso en 90% de los casos. Se utiliza la guía ultrasonográfica o de tomografía computadorizada para la aspiración del absceso.El metronidazol es el agente amebicida de preferencia en el tratamiento del absceso amibiano del hígado. El drenaje abierto está contraindicado. En los casos en que falla la terapia con amibicidos, se realiza el drenaje cerrado guiado por US o por TC. La infección secundaria de un absceso amibiano del hígado es un fenómeno extraordinariamente raro.La identificatión y determinatión de la sensibilidad antibiótica de los microorganismos responsables del absceso piógeno representa un paso crucial en su manejo. A menos que se haga necesario realizar una laparotomía para la correción del algún proceso intraabdominal o porque el absceso es excesivamente grande, el tratamiento inicial del absceso piógeno es un ciclo de antibióticos propiados de 2 semanas, seguidos de tratamiento con antibióticos orales por un mes. La mayoría de los abscesos piógenos del hígado responde a este tipo de tratamiento. Si se requiere drenaje de un absceso piógeno, la técnica de preferencia es la punción percutánea por medio de un catéter guiado por US o TC. El drenaje quirúrgico abierto debe reservarse para aquellos casos en que la laparatomía es necesaria por razones diferentes o en que hay falla en la respuesta a un ciclo de terapia antibiótica adecuada y el drenaje percutáneo no es factible.La mortalidad en el manejo del absceso amibiano del hígado debe ser nula, y para el absceso piógeno de menos de 10%.

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Résumé L'abcès hépatique amibien ou à pyogènes peut être diagnostiqué avec une grande précision soit par l'échographie, soit par la tomodensitométrie. L'échographie est la méthode de choix et détecte presque 100% des abcès. On obtient la confirmation du diagnostic d'abcès amibien par le test d'hémagglutination indirecte qui est positive dans presque 100% des cas. On doit faire des cultures de pus provenan de l'abcès et des hémocultures en cas d'abcès à pyogènes du foie. Ces cultures ont été positives dans 90% des cas. L'échographie et la tomodensitométrie aident à guider le drainage de abcès.Dans le traitement de l'abcès amibien du foie, le métronidazole est l'amibicide de choix. Le drainage à ciel ouvert est contreindiqué. Pour les cas qui ne répondent pas aux amibicides, un drainage percutané guidé par la tomodensitométrie ou l'échographie est indiqué. La surinfection d'un abcès amibien du foie est extrêmement rare.L'identification et la détermination de la sensibilité aux antibiotiques des organismes responsables de l'abcès à pyogènes est une étape extrêmement importante. A moins qu'une laparotomie soit nécessaire pour traiter une infection intraabdominale associée ou que le volume de l'abcès soit extrêmement important, le traitement initial d'un abcès à pyogènes comprend 2 semaines d'antibiotiques adaptés par voie générale suivies d'un mois d'antibiotiques par voie orale. La plupart des abcès à pyogènes répondront bien à ce traitement. Si le drainage d'un abcès à pyogènes s'avère nécessaire, la meilleure technique est percutanée avec un cathéter inséré sous contrôle tomodensitométrique ou échographique. On réservera le drainage chirurgical à ciel ouvert aux cas où une laparotomie est nécessaire pour d'autres raisons et où le malade n'a pas répondu à l'antibiothérapie adaptée et chez qui le drainage percutané est impossible à faire.La mortalité de l'abcès amibien traité devrait approcher 0% et atteindre pour l'abcès à pyogènes moins de 10%.

     

The impact of the “cisplatin era” of treatment on survival in testicular cancer

Summary The records of all testicular cancer patients evaluated and treated at our medical center during two consecutive 9-year periods were reviewed and analyzed for prognostic factors, particularly the impact of cisplatin-based combination chemotherapy. The data base of 244 patients was divided into two eras: 1970–1978, defined as the pre-cisplatin era (n=101) and 1979–1987, the cisplatin era (n=143). Statistically improved survival (P=0.024) was noted for the 165 nonseminoma patients and for a grouping of 143 patients treated with combination chemotherapy (P=0.004) during the cisplatin era. Stratification by stage revealed that stage II patients had the most significant survival advantage (P=0.001) during the cisplatin era; cancer mortality improved from 48% to 9%. Cancer death rates for stage III patients decreased from 58% to 39% which is clinically but not statistically significant (P=0.497). Stage I patients and the seminoma population did well during both eras, and the impact of cisplatin could not be statistically confirmed in this study for these subgroups. Multivariate statistical analysis confirmed the importance of the era of treatment for the nonseminoma population.     

Using neural networks to diagnose cancer

While artificial brains are in the realm of science fiction, artificial neural networks (ANNs) are scientific facts. An artificial neural network is a computational structure modeled somewhat on the neural structure of the brain; both have many highly interconnected processing elements. These biologically inspired processing elements are taught by feeding examples until the results are acceptable. In the past 5 years, neural networks have become successful in providing meaningful second opinions in clinical diagnosis. In our research, a prototype artificial neural network was trained on numeral ultrasound data of 52 actual cases and then correctly identified renal cell carcinoma from renal cysts and other conditions without diagnostic errors. Our nonlinear artificial neural network was trained on software using the standard backpropagation paradigm on a 80386 microcomputer. Our ANN learned from ultrasound data in 52 cases (17 malignant, 30 cysts, and 5 other) at a Memphis hospital. The trained prototype performed without error on 47 cases which were not in the data used for training. This prototype must be validated by extending this study to more cases.     

Cardiac sensitization induced by phenobarbitone and prolonged by CS2

The arrhythmogenic effect of 8 g/kg noradrenaline given i.v. was increased in male rats pretreated 1–2 day s earlier with phenobarbitone and starved from the time of the first phenobarbitone injection (80 mg/kg followed by 50 mg/kg 6 h later). Daily exposure to 4.0 mg/l CS₂ (first exposure 24 h after the first phenobarbitone injection) for 4 h prevented the decline in susceptibility on the 3rd and 4th days after phenobarbitone, when the reaction of unexposed rats to noradrenaline returned to normal.     

A Phase II trial of flavopiridol (NSC #649890) in patients with previously untreated metastatic androgen-independent prostate cancer.

PURPOSE: Flavopiridol is a cyclin-dependent kinase inhibitor with preclinical activity against prostate cancer cell lines. A Phase II trial was conducted to determine the activity of flavopiridol in patients with metastatic hormone-refractory prostate cancer. EXPERIMENTAL DESIGN: A total of 36 patients was enrolled from several institutions and treated with a 72-h continuous infusion of flavopiridol every 14 days at the eventual starting dose of 40 mg/m(2)/day. Dose escalation up to 60 mg/m(2)/day was permitted if no significant toxicity was observed. Responses were assessed every 12 weeks. Only those patients completing four courses of the 72-h infusion were considered evaluable for response because the primary objective was to determine progression-free survival at 6 months given the cytostatic nature of the agent. RESULTS: This study was conducted in a two-stage fashion. During the first stage, at least 20 evaluable patients needed to be enrolled to assess response. There were 22 of 36 patients evaluable for response. No objective responses were observed. Only 4 patients had stable disease for 16, 26, 29, and 48 weeks, respectively, stopping the trial by design as only 3 of 22 (14%) of the patients met the 6-month progression-free survival end point. The most common toxicities were diarrhea (grade 1 and 2) and nausea, although some grade 3 and 4 diarrhea (11 and 6%, respectively) were evident. CONCLUSIONS: Flavopiridol has disappointing single-agent activity in hormone-refractory prostate cancer when administered at this dose and schedule. Its use in prostate cancer should be reserved for evaluation in combination therapies or alternative schedules.     

Lamivudine for the prevention of hepatitis B virus reactivation in hepatitis B s-antigen seropositive cancer patients undergoing cytotoxic chemotherapy.

PURPOSE: For cancer patients receiving cytotoxic chemotherapy, hepatitis B virus (HBV) reactivation is a well described complication resulting in varying degrees of liver damage. The objectives of this study were to assess the efficacy of the antiviral agent lamivudine in reducing the incidence of HBV reactivation and diminishing morbidity and mortality of cancer patients with chronic HBV infection during chemotherapy. PATIENTS AND METHODS: Two groups were compared in this nonrandomized study. The prophylactic lamivudine group consisted of 65 patients in a phase II study who were treated with lamivudine before and until 8 weeks after discontinuing chemotherapy. The historical controls consisted of 193 consecutive patients who underwent chemotherapy without prophylactic lamivudine. Significant prognosticators for the development of HBV reactivation were determined based on data from the controls. Potential confounding factors were identified between the two groups. The outcomes were compared. RESULTS: In the controls, lymphoma and anthracycline usage were factors identified to be associated with reactivation. The two groups were comparable in most baseline characteristics, although in the prophylactic lamivudine group, there were significantly more patients with lymphoma and receiving anthracyclines. In the prophylactic lamivudine group, there was significantly less HBV reactivation (4.6% v 24.4% in the controls; P <.001), fewer incidences of hepatitis (17.5% v 44.6%; P <.0001) that were less severe (4.8% v 18.7%; P =.0005), and less disruption of chemotherapy (15.4% v 34.6%; P =.0029). The reduction in overall mortality was not statistically different. CONCLUSION: Prophylactic lamivudine significantly reduced the incidence of HBV reactivation and the overall morbidity of cancer patients undergoing chemotherapy.