Antibody responses to Bacillus Calmette-Guérin during immunotherapy in bladder cancer patients.

Levels of serum antibody to Bacillus Calmette-Guérin (BCG) were determined by solid-phase radioimmunoassays in 48 normal donors and 60 patients with bladder cancer. Of 57 patients enrolled in a randomized prospective controlled trial of BCG immunotherapy, 47 were followed for up to 30 months, thus permitting comparisons of tumor recurrence, delayed cutaneous hypersensitivity responses to purified protein derivative (PPD), and serum BCG antibody levels at specific intervals during the clinical course. Sera from normal donors and cancer patients prior to BCG therapy had equally low levels of BCG antibody. AFter administration of intravesical and percutaneous BCG, significant rises of serum BCG antibody levels were detected in 23 of 24 randomized BCG immunotherapy patients. Skin test responses to PPD and serum BCG antibody levels had a close correlation as immune response indicators in 14 of 24 BCG therapy patients, while rises in serum BCG antibody levels were a better response indicator than PPD skin test reactions in the other 10 patients. Eleven of the 23 patients randomized into the non-BCG treatment group had tumor recurrence, although tumors recurred in only six of the 24 randomized patients in the BCG therapy group. Two additional nonrandomized BCG-treated patients had tumor recurrence. All eight BCG-treated patients with tumor recurrence had documented increases in serum BCG antibody levels after BCG therapy. Only three of these eight patients had conversion of PPD skin test responses from negative to positive; three were positive before immunotherapy and two remained negative after BCG therapy. Levels of antibodies reactive with human adenovirus type 5 and with Escherichia coli antigens were similar in sera from normal donors and from the randomized bladder cancer patients in both the BCG and non-BCG treatment groups. These results suggest that serum BCG antibody responses are as useful as PPD skin tests in identifying immunological responses to the immunoadjuvant BCG during immunotherapy trials in cancer patients.

Transferrin binding to two human colon carcinoma cell lines: characterization and effect of 60-Hz electromagnetic fields.

125I-Labeled human transferrin was used to study the binding of transferrin to Colo 320 DM and Colo 205 human cell lines derived from adenocarcinomas of the colon. Although transferrin uptake was greater in both cases at 37 degrees than at 4 degrees it was found that slightly greater than two-thirds of the transferrin associated with the cells at 37 degrees was not bound to surface receptors but rather had been internalized by the cells. Subsequent analysis of true surface binding at 4 degrees by Scatchard analysis allowed determination of the number of transferrin receptors as well as association constants for the interaction. The number of transferrin receptors per cell was found to be inversely related to the cell density of the cultures from which cells were removed for study. Association constants were unaffected by cell density, with average values of 1.2 and 5.4 X 10(8) M-1 obtained for Colo 320 DM and Colo 205, respectively. Additionally, maximum theoretical numbers of receptors of 1.05 X 10(5)/cell for Colo 320 DM and 1.39 X 10(5)/cell for Colo 205 were determined. Furthermore, exposure of Colo 205 cells to three different experimental situations, i.e., 60 Hz-generated electric field only (E+, 300 mA/m2rms), magnetic field only (M+, 1.0 gauss rms), and combined electric + magnetic fields at these intensities (E+M+), altered the expression of transferrin receptors as compared to a concurrently run unexposed control population of cells (E-M-). In three separate experiments the number of transferrin receptors quantitated on both M+ and E+M+ cells was independent of cell culture density and was close to or exceeded the maximum theoretical number of receptors determined for this cell line. In contrast, E+ cells expressed fewer transferrin receptors than was predicted on the basis of cell culture density.

Comparison of isomers of ketamine on catalepsy in the rat and electrical activity of the brain and behavior in the cat.

The present study compared the relative potency and efficacy of the two isomers of ketamine on the duration of catalepsy (loss of righting reflex) in female rats and on the behavior and electroencephalogram of cats. In the rat, at small doses, the S(+) isomer was more potent than the R(-) isomer or racemic ketamine, while at larger doses, the S(+) isomer and the racemate were equipotent and the R(-) isomer was significantly less potent. Tolerance developed rapidly to the effects of either isomer and both were equally cross-tolerant to racemic ketamine. Sub-effective doses of morphine significantly increased the potency of S(+), R(-) and racemic ketamine on the duration of catalepsy. Sub-effective doses of either isomer augmented the duration of catalepsy, induced by small doses of morphine, but reduced that of large doses. In cats, there was a parallel time course and progression of behavioral and electroencephalographic states in response to equal total doses of either racemic ketamine, an artificial 50:50 mixture of S(+) and R(-) isomers, or the S(+) isomer alone; approximately equivalent effects required twice the dose of the R(-) isomer. It is concluded that there is a common site of action for the two isomers, but there is also a stereospecific difference in potency, as regards the induction of catalepsy in the rat and behavioral and electroencephalographic effects in the cat. Stereospecificity was not apparent in the development of tolerance, cross-tolerance or the augmentation of the response to morphine.

Catalepsy induced by combinations of ketamine and morphine: potentiation, antagonism, tolerance and cross-tolerance in the rat.

Previous studies demonstrated that both ketamine and morphine induced analgesia and catalepsy in the rat. Pre-treatment with ketamine produced cross-tolerance to morphine, whereas pretreatment with morphine did not induce cross-tolerance to ketamine but rather augmented the cataleptic response; this augmentation was attributed to residual morphine in the brain. The present studies explored the duration of the loss of righting reflex induced by sub-effective doses of ketamine and morphine, administered simultaneously. There was mutual potentiation between sub-effective doses of ketamine and morphine, but sub-effective doses of ketamine partly antagonized fully-effective doses of morphine. Latency to the loss of righting reflex, rigidity and behavior on recovery, reflected the relative predominance of ketamine or morphine in each combination. Naloxone inhibited the induced cataleptic effects. The degree and time course of development of tolerance to daily administration of sub-effective dose combinations of ketamine and morphine were similar. Rats, tolerant to ketamine-dominant combinations, were cross-tolerant to both drugs, while those tolerant to morphine-dominant combinations were cross-tolerant to morphine but showed either no cross-tolerance or an augmented response to ketamine. While the mutual potentiation, antagonism and tolerance suggest common mechanisms for the induced catalepsy, differences in latency, rigidity and behavior, asymmetry of cross-tolerance and a widely-different ID50 for naloxone would argue against an action at a single opioid site.

Synthetic opioids compared with morphine and ketamine: catalepsy, cross-tolerance and interactions in the rat.

Previously it has been shown in rats that both ketamine and morphine induced analgesia and, at larger doses, catalepsy and loss of the righting reflex, all of which were reversed by naloxone at widely different doses. Tolerance developed rapidly to either ketamine or morphine and there was cross-tolerance from ketamine to morphine. However, morphine potentiated the cataleptic effect of ketamine, whether fully-effective doses of morphine were given before ketamine or subeffective doses of both were given concurrently. The present study extends these observations to three specific mu-receptor agonists (sufentanil, fentanyl and alfentanil) and two mu- and kappa-agonist, mu-antagonist opioids (nalbuphine and butorphanol). All five of these opioids potentiated the cataleptic effect of ketamine. Each of the three specific mu agonists showed rapid development of tolerance. Fentanyl and alfentanil showed mutual cross-tolerance with ketamine, but sufentanil did not. This lack of sufentanil-ketamine cross-tolerance may reflect separation of the sites of agonist action and the sites of development of tolerance for the opioids and for ketamine. The potentiating effects of nalbuphine and butorphanol suggest that they potentiate ketamine-induced catalepsy, either by kappa-receptor interactions or by a mu agonist effect. It is suggested that the cataleptic effect of a combination of individually-subeffective doses of ketamine and morphine, rather than ketamine and one of the synthetic opioids, might be of more potential clinical usefulness.

Seasonal and sex influences on ketamine-induced analgesia and catalepsy in the rat; a possible role for melatonin.

Data from this laboratory on ketamine-induced analgesia and catalepsy in rats revealed that factors other than dose modified the difference in the latency of the tail flick response (TFLD), a measure of analgesia, and the duration of the loss of the righting reflex (DLRR), a measure of catalepsy. Untreated female rats showed a longer latency than males in their response to a noxious stimulus at midnight, but not at noon. Females also showed a longer loss of righting reflex response to ketamine than did males, whether at noon or midnight; the loss of righting reflex at night was augmented in both. Although females showed analgesia with administration of ketamine at doses smaller than those which induced catatonia, males showed no analgesia without catatonia and comparable loss of the righting reflex occurred at doses much larger than for females. There was a 3-fold increase in the latency of the tail flick response and loss of righting reflex during the winter, as compared with summer, for females treated with ketamine; males showed a similar variation in the loss of righting reflex. Since analgesia is produced by both melatonin and ketamine, and since ketamine appears to share opiate receptors with an endogenous ligand, beta-endorphin, a role was sought for the pineal and melatonin in the variation of responsiveness to ketamine. Pinealectomized rats failed to show augmentation of the loss of righting reflex induced by ketamine at night and mice showed a seasonal variation in the analgesia induced by melatonin.(ABSTRACT TRUNCATED AT 250 WORDS)

A new perspective of microbial survival and dissemination in a prospectively contaminated air-fluidized bed model.

A major concern of users of air-fluidized beds has been the possibility that such beds might be a source of microbial contamination. The purpose of this series of prospective, controlled experiments was to measure quantitatively the dissemination and survival of Bacillus subtilis, a species of bacterium that forms desiccation-resistant spores, as it was associated with circulating and clumped microbeads after challenge in an air-fluidized bed operating under decontamination conditions of heating at 48 degrees C and microbead agitation by an air flow of 100% at 110 cu ft/min. Microbead samples collected after B. subtilis challenge from predesignated depths and locations within the air-fluidized bed at 0.25, 1, 2, 4, 24, and 48 hours were assayed for colony-forming units (CFU) of challenge bacteria by end point dilution and streak-plate assays. Results of three experiments with an average challenge of 1110 CFU of B. subtilis per gram of microbeads indicated that no more than 0.46 CFU of B. subtilis per gram of circulating microbeads and 0.78 CFU per gram of clumped microbeads were detected after 24 to 48 hours at decontamination conditions. Few microbes were detected during three control (sterile water) challenges of the air-fluidized bed operating at 33 degrees C with 95% to 100% air flow. This study has demonstrated that an air-fluidized bed operating at the described decontamination conditions for 24 to 48 hours caused significant decreases, averaging a thousandfold per gram, in levels of microbead-associated challenge bacteria.(ABSTRACT TRUNCATED AT 250 WORDS)

A prospective study of femoral catheter-related thrombosis in children.

OBJECTIVES: To establish the incidence and correlate clinical findings of femoral venous catheter-related thrombus formation in critically ill children. RESEARCH DESIGN: Observational prospective blinded study. SETTING: University-affiliated pediatric hospital intensive care unit. PATIENTS: Twenty children admitted to the pediatric intensive care unit who had percutaneous femoral venous catheters placed while in the pediatric intensive care unit. INTERVENTIONS: None. MEASUREMENTS: Duplex Doppler ultrasonography evaluation of femoral vein catheters at 1 to 2, 3 to 5, and 7 to 10 days after placement was used to detect the presence of thrombus formation and venous occlusion. Demographic patient data, pediatric risk of mortality scores, and clinical findings, including leg swelling and whether catheters would aspirate blood, were also recorded. Continuous data were analyzed using the Mann-Whitney U Test, and categorical data were compared with Fisher's Exact Test. Statistical significance was assigned at a P value of .05 or less. CONCLUSIONS: The overall incidence of catheter-related femoral vein thrombus formation was 35% (7/20). Ipsilateral leg swelling and the inability to aspirate blood from the catheter were significantly associated with thrombus formation. Patients who developed thrombi were younger and smaller than those who did not. In six of seven patients, thrombus formation was clinically occult when first demonstrated by ultrasonography.

Sensitivity of human oral tumor cells to human adenovirus.

Human cells in early passage cultures of benign oral tumors, normal oral tissues, normal lung tissues and, especially, in long-term established oral carcinoma cultures were highly susceptible to infection by human adenovirus types 5, 21, and 31. In contrast, replication of each adenovirus type was markedly limited in inoculated cells of newly-established oral squamous cell carcinoma cultures.

Humoral immune responses to adenoviruses, herpes virus type 1, and Candida albicans in sera of dental patients with oral neoplastic and periodontal diseases.

Levels of immunoglobulin class-specific antibodies as determined by solid phase radioimmunoassays to herpes simplex virus type 1 (HSV-1), human adenovirus types 5, 21, and 31 and to Candida albicans in sera from untreated healthy dental patients were not significantly different from levels of these antibodies in sera from untreated dental patients with benign oral tumors, oral carcinoma, or periodontal disease. These results show that higher levels of immunoglobulin class-specific antibodies to HSV-1, the three adenoviruses, or Candida albicans are not a consistent finding in sera from patients with oral cancer when comparisons are made with healthy patients and patients with other oral diseases.

Congenital masses of the lung: prenatal and postnatal imaging evaluation.

Congenital masses of the lung are a spectrum of interrelated abnormalities that includes congenital lobar overinflation, bronchogenic cyst, congenital cystic adenomatoid malformation (CCAM) and sequestration. The prenatal and postnatal imaging features of these lesions are reviewed, emphasizing the importance of serial prenatal sonograms and postnatal imaging studies, including radiography and computed tomography. Masses that become inconspicuous, or disappear on serial prenatal sonograms are discussed, as well as the importance of postnatal imaging studies in the evaluation of these lesions. Finally, the management of congenital masses of the lung is reviewed, emphasizing the importance of imaging studies in the preoperative evaluation.

Beneficial effect of Aloe on wound healing in an excisional wound model.

Recent evidence from in vitro and in vivo experiments suggests that topical antimicrobials may be toxic to fibroblasts and keratinocytes and retard wound healing. The purpose of this study was to determine the effects of Aloe, a potential wound-healing agent, on wound contraction in excisional wounds treated with topical antimicrobials. Sprague-Dawley rats were prepared with four 1.5 cm2 dorsal defects through the skin and panniculus. The animals were divided into five groups (n = 10 per group): (1) Aloe, (2) NaOCl solution (0.025%), (3) mafenide acetate, (4) mafenide acetate + Aloe, and (5) control. Wounds were treated topically for 14 days 3 times a day. Serial standard photographs and serial wound planimetry were performed weekly. Following healing, the breaking strength of each resultant scar was determined using an Instron tensiometer. Kruskal-Wallis, ANOVA, and multiple comparison methods were used for data analysis. Aloe and NaOCl solution significantly accelerated wound contraction (p < 0.05). In the mafenide acetate + Aloe group, contraction was similar to the control, whereas the mafenide acetate alone retarded wound healing. The addition of Aloe in combination and alone in wounds increased the breaking energy when compared to controls (p < 0.05). Aloe appears to expedite wound contraction and neutralize the wound retardant effect seen with the topical mafenide acetate alone. This effect appears to be due to an increased collagen activity, which is enhanced by a lectin, consequently improving the collagen matrix and enhancing the breaking strength.

In vitro immune monitoring of antibody response in dogs given chemoimmunotherapy for lymphoma.

Clinical remission in 30 dogs with lymphoma was induced with a combination of vincristine, L-asparaginase, cyclophosphamide, and doxorubicin HCl, administered sequentially, and then an autochthonous tumor cell vaccine, given intralymphatically, as maintenance therapy. Humoral antibody amounts were monitored in 11 dogs, using a solid-phase bead-type radioimmunoassay. The median survival of the 30 dogs was 13 months from the start of chemotherapy (range, 7 to 25 months; mean, 13.8). The median remission duration was 16 weeks (range, 9 to 98 weeks; mean, 26.8). Correlation between increase in amount of humoral antibody was significant (P = 0.0001 to 0.012), before and after chemoimmunotherapy, in dogs responding to therapy, compared with that in dogs not responding to therapy.

Human adenovirus antibody in sera of normal and tumour-bearing dogs.

Serum specimens from 42 normal dogs and 42 with untreated malignant tumours were assayed for the presence of antibodies to human adenovirus types 5, 21 and 31 and to infectious canine hepatitis (ICH) virus. Radioimmunoassays using human adenovirus antigens showed that 71 per cent (30/42) of all dogs with tumours, but only 19 per cent (8/42) of all normal dogs, were positive for human adenovirus antibody. Canine sera reactive with antigens of one human adenovirus type in radioimmunoassays were also reactive with antigens of the other two types. Dogs bearing malignant lymphoma or squamous cell carcinoma tumours had higher levels of antibody against adenovirus type 5 antigens. Human adenovirus type 5 was neutralised by sera from four tumour-bearing and two normal dogs, while sera from two normal and five tumour-bearing dogs were positive in immunodiffusion tests with human adenovirus antigens. Levels of ICH antibody in sera of normal adult dogs and adult dogs with tumours were not markedly different when measured by radioimmunoassays. Likewise, sera from these two groups of dogs had similar ranges of ICH neutralising antibody titres. In contrast, levels of ICH antibody detected by the serological assays in sera from non-pet, non-vaccinated pups were either markedly low or absent. Possible explanations for the observed increased levels of human adenovirus antibody in sera of tumour-bearing canine pets are discussed.

Time dependent decreases of maternal canine virus antibodies in newborn pups.

Levels of passively transferred maternal antibodies to three canine viruses, rabies virus (RV), canine distemper virus (CDV) and infectious canine hepatitis (ICH) virus, in serum specimens from 14 fetal pups and in serial serum specimens collected up to 45 days after whelping from 14 neonate pups were compared with levels of antibodies to these viruses in milk and sera collected concurrently from their respective dams. Radioimmunoassays using RV-, CDV- and ICH virus-specific antigens showed that sera from all fetal pups had detectable levels of antibodies to all three canine viruses and ICH neutralising antibodies were detected in sera from 10 of the 14 fetal pups. As the time after whelping increased, titres of RV-, CDV- and ICH virus antibodies measured by radioimmunoassay and ICH virus neutralising antibody tests in serially collected specimens of milk from dams rapidly decreased, while titres of the antibodies in serum specimens from newborn pups in their litters steadily decreased. Individual fetal and newborn pups with a high titre of antibody to one virus also had high titres to the other two viruses, although a wide range of titres was observed among pups in each of the litters studied. Markedly higher titres of antibody to all three viruses were observed in serially collected specimens of sera from dams than in sera from fetal and newborn pups in their litters. Results show that maternal RV, CDV and ICH virus antibodies are transferred from dams to pups in utero and by nursing. Levels of these maternal virus-specific antibodies in newborn pup sera decreased at similar rates as time after whelping increased.

Retardation of embryogenesis by extremely low frequency 60 Hz electromagnetic fields.

Fertilized Medaka fish eggs were used to determine if electromagnetic fields, designed to simulate those beneath a high voltage power line, have biological effects on vertebrate embryo development. The newly fertilized eggs were exposed to a 60 Hz electrical field of 300 mA/m2 current density, a 60 Hz magnetic field of 1.0 gauss RMS, or to the combined electric plus magnetic fields for 48 hours. No gross abnormalities were observed in any of the embryos as they developed, but significant development delays were seen in those embryos exposed to either the magnetic or to the combined electromagnetic fields; delays were not seen in the embryos exposed to the electrical field. Thus, a 60 Hz magnetic field like that encountered in a man made powerline environment was shown to retard development of fish embryos.

Effect of radiofrequency radiation on mRNA expression in cultured rodent cells.

Four rodent cell lines were exposed to 2450 MHz microwave radiation at a Specific Absorption Rate (SAR) of 103.5 +/- 4.2 W/kg for varying lengths of time at 37 degrees, 40 degrees, 42 degrees and 45 degrees C. mRNA was extracted from microwave-exposed and sham-exposed cells and dot blotted or Northern blotted to nitrocellulose. Radioisotope labelled DNA probes of oncogenes, heat shock protein or long terminal repeat sequences were hybridized to the mRNA, and the resulting autoradiographs analyzed for differences in levels of mRNA expression between exposed and nonexposed samples. With the cell lines and probes used in this study no significant differences in mRNA expression were observed after microwave exposure.