Intratumoral differences in methotrexate levels within human osteosarcoma xenografts studied by microdialysis

A central tenet in oncology is the assumed relationship between drug concentration and cytotoxicity. Determinations of drug levels in tumor tissues are, however, generally not undertaken. Microdialysis is a method where continuous drug monitoring may be achieved by sampling of low molecular weight substances from the extracellular space. By employing this technique it is possible to observe variable drug levels within tissues, including tumors, over time. Herein, we present results from a nude rat model where subcutaneous human osteosarcoma xenografts were established prior to the administration of the antifolate methotrexate as an intravenous infusion. Significant differences in drug exposure within single tumors were evident. Generally, peak drug concentrations were lower and drug efflux slower from the center of the tumors as compared to the periphery. The use of microdialysis could be an important tool for optimizing current strategies in anticancer chemotherapy.     

Real-time enzyme kinetics monitored by dual-color fluorescence cross-correlation spectroscopy

A method for sensitively monitoring enzyme kinetics and activities by using dual-color fluorescence cross-correlation spectroscopy is described. This universal method enables the development of highly sensitive and precise assays for real-time kinetic analyses of any catalyzed cleavage or addition reaction, where a chemical linkage is formed or cleaved through an enzyme's action between two fluorophores that can be discriminated spectrally. In this work, a homogeneous assay with restriction endonuclease EcoRI and a 66-bp double-stranded DNA containing the GAATTC recognition site and fluorophores at each 5' end is described. The enzyme activity can be quantified down to the low picomolar range (>1.6 pM) where the rate constants are linearly dependent on the enzyme concentrations over two orders of magnitude. Furthermore, the reactions were monitored on-line at various initial substrate concentrations in the nanomolar range, and the reaction rates were clearly represented by the Michaelis-Menten equation with a KM of 14 +/- 1 nM and a kcat of 4.6 +/- 0.2 min-1. In addition to kinetic studies and activity determinations, it is proposed that enzyme assays based on the dual-color fluorescence cross-correlation spectroscopy will be very useful for high-throughput screening and evolutionary biotechnology.     

Augmenting the immunogenicity of synthetic MUC1 peptide vaccines in mice

Human mucin MUC1 is abundantly expressed in some cancers of epithelia] origin and is largely restricted to the apical surface of secretory cells in normal tissues. It is, therefore, a potential target for cancer immunotherapy. In preparation for clinical trials, vaccines containing synthetic MUC1 peptides of different lengths and sequences mixed with various adjuvants or covalently attached, using different linker methods, to protein carrier keyhole limpet hemocyanin (KLH) were studied in mice. MUC1 peptides (containing 30 amino acids), plus adjuvants QS-21 or Bacillus Calmette-Guerin, were incapable of inducing antibody. However, MUC1 peptides conjugated to KLH (MUCl-KLH), plus QS-21, induced high titer antibody against the immunizing peptides and against MUC1-expressing tumor cells. Although T-cell responses, including delayed-type hypersensitivity, lymphocyte proliferation, and CTL, were not observed in mice immunized with these vaccines, significant protection from MUC1-expressing tumor cell challenge in mice immunized with MUC1-KLH was observed. Based on these studies, a vaccine containing MUC1-KLH conjugate prepared with m-maleimidobenzoyl-N-hydroxysuccinimide ester linker, plus QS-21, has been constructed for testing in clinical trials.     

Relationship between prophylactic effect of lithium therapy and family history of affective disorders

Lithium therapy response and age of onset (AOO) were studied in 98 patients with bipolar affective disorder (BPAD) who were divided into subgroups depending on type of family history of affective disorders. The highest (33.0 years) and lowest (25.5 years) age of onset were found in nonfamilial patients and in familial patients with a first-degree relative of BPAD, respectively. Nonfamilial patients showed the best response to lithium. There were 0.9 episodes/year off lithium compared to 0.3 episodes/year on lithium (an 88% decrease). A poorer response (a 71% decrease; a reduction from 1.39 episodes per year off lithium to 0.65 on lithium) was found in familial patients with a first-degree relative of BPAD. Differences in serum lithium values between the groups could not explain the observed differences. Thus, familial patients showed a more severe manifestation of the disease with an earlier AOO and a lower prophylactic effect of lithium.     

Diagnosis of phenylketonuria in a 35-year-old mother in relation to prenatal diagnosis of intrauterine growth retardation with microcephaly

BACKGROUND: Some French pregnant women with phenylketonuria (PKU), born before 1978, have not been tested with the Guthrie method during the neonatal period. They have a risk of spontaneous abortion and their infants are often mentally retarded with microcephaly and/or congenital heart anomaly. CASE REPORT: A woman with a moderate mental retardation became pregnant at the age of 31 years. Her newborn had a severe intrauterine growth retardation with microcephaly and developed mental retardation. This mother became pregnant again 4 years later. Repeated fetal ultrasonography showed progressive growth retardation. Maternal blood phenylalanine concentration was 18 mg/100 mL. Therapeutic abortion at 27 weeks of gestational age showed a fetus with several abnormalities, particularly cardiovascular and cerebral. CONCLUSION: It is still possible to meet women with unrecognized atypical PKU. Fetus or infant born with unexplained growth retardation and microcephaly requires search for maternal PKU.     

Pneumatic dilatation is effective long-term treatment for achalasia

Although pneumatic dilatation (PD) has been an established treatment for achalasia for decades, there is limited information on its long-term clinical efficacy. We have followed up the clinical status of patients having PD with a 30- or 35-mm balloon by one of us (D.O.C.) over a 25-year period. Of 144 patients whose initial records were available for review, 31 could not be contacted. Of the remaining 113 patients, 72 (64%) responded to a questionnaire assessing swallowing status and patient satisfaction, and this forms the basis of this report. There were 32 men and 40 women, with mean age 46 years (range: 17-78); mean length of follow-up since PD was 6.5 years (range: 10 months to 25 years). Success was primarily defined by the need for no additional therapy for achalasia other than one or two PD's. PD was effective long-term treatment in 61/72 patients (85%); only four of these required a second PD over this time interval. There was no significant difference in any of the following parameters between patients with a treatment success or failure: age, sex, size of pneumatic dilator, and duration of symptoms prior to PD. Response was significantly better (P < 0.05) in patients having no prior dilatation (43/47; 91%) than in those in whom another physician had performed prior dilatation (18/25; 72%). In response to the question of whether they would select PD again, 68 patients (94%) responded positively. In conclusion, pneumatic dilatation performed using a consistent technique by an experienced physician is effective long-term therapy for achalasia patients of all ages. Most patients require only one dilatation.     

Characterization of a mouse monoclonal IgG3 antibody to the tumor-associated globo H structure produced by immunization with a synthetic glycoconjugate

OBJECTIVE: The objective of this study was to assess the degree of BP Tracking from childhood to adulthood and to evaluate whether high BP levels persist over time and progress to adult hypertension. PATIENTS AND METHODS: Two hundred and twenty-two healthy schoolchildren living in the North of Portugal were assessed at 17 year intervals, starting in 1979 (cohort 1) aged 5 to 18 years, and again in 1996 (cohort 2). Tracking indices (Ti) were calculated as follows: Ti = (2x + y-z) /N/0.89, where x, y and z refer to the total number in the same, adjacent and remote trisections, respectively, and N = x + y + z. If Ti > 1 there is positive tracking. RESULTS AND CONCLUSIONS: For systolic and diastolic blood pressure, all Ti were greater than 1.0. All individuals that remained in the 3rd tertil, 17 years later, weigh more and are more obese than those of the 1st tertil. 56.6% of the individuals that belong to the 3rd tertil are now hypertensive, which means that a significant percentage of the children with high blood pressure in the first survey will be hypertensive in the future.