Immunoglobulin E antibodies from pancreatic cancer patients mediate antibody-dependent cell-mediated cytotoxicity against pancreatic cancer cells.

In addition to allergy and parasitic infections, immunoglobulin E (IgE) has been shown recently to possess anti-viral and anti-cancer effects. We investigated serum levels of IgE, its low-affinity receptor, soluble CD23 (sCD23) in patients with pancreatic cancer and the effect of IgE against pancreatic cancer cells. Twelve patients were evaluated for pancreatic cancer by imaging and confirmed by biopsy. Fifteen healthy volunteers served as controls. Serum Igs (IgG, IgM, IgA, IgE) and sCD23 levels were determined (enzyme-linked immunosorbent assay, nephelometry) and the presence of cancer-specific IgE was assessed (fluorescence microscopy, Western blot). IgE anti-cancer activity was determined by antibody-dependent cell-mediated cytotoxicity (ADCC). Serum levels of IgE and sCD23 were elevated significantly in patients with pancreatic cancer versus controls, whereas no differences were observed in other Ig isotypes (IgG, IgM, IgA). Flow cytometry and immunofluorescence microscopy demonstrated similar presence of IgG and IgE pancreatic cancer Igs. However, Western blot analysis indicated differences in IgG and IgE antigen-specific antibodies; IgE antibody recognized a 50 kD protein. ADCC studies demonstrated that serum and purified IgE-mediated cytotoxicity against pancreatic cancer cells, effects which were reversed with anti-IgE neutralizing antibody and IgE depletion (immunoaffinity); greater cytotoxicity was observed in patient serum when compared with healthy controls. These data suggest that IgE and sCD23 may serve as useful biomarkers for patients with pancreatic cancer and may be important in the immune response to this disease in that IgE-directed therapy may help to direct treatment.

Evaluation of graft-host response for various tissue sources and animal models.

The efficacy of pancreatic islet transplants in correcting hyperglycemia and slowing the progression of complications in diabetics has been confirmed by many experimental and clinical studies. Unfortunately, the availability of human islets is extremely limited and, therefore, treatment of large numbers of human diabetic patients will almost certainly require either the use of islets harvested from animals (xenografts) or the use of insulin-secreting genetically modified cells of either human or animal origin. There is currently no effective regimen which will allow long-term survival of xenogeneic islets from widely unrelated donor-recipient combinations, such as pig-to-rodent, pig-to-dog, or pig-to-primate. There is considerable interest in the development of immunoisolation techniques for protection of donor islets. However, most materials used in immunoisolation devices are relatively bio-incompatible. Poly-L-lysine-alginate microcapsules are biocompatible and provide an optimal geometry for transmembrane diffusion of insulin and nutrients. Microcapsules allow long-term survival of xenogeneic islets in diabetic rodents or dogs with induced diabetes. However, mice and rats with spontaneous diabetes destroy encapsulated islet grafts within 2 to 3 weeks. Biopsies reveal large numbers of macrophages, immunoglobulins and limited numbers of helper and cytotoxic T-cells in the peri-microcapsule environment of the peritoneal cavity. Cytokines have been identified in peritoneal fluid from mice with islet grafts and may play a role in encapsulated islet destruction. Targeted immunomodulation by treatment of recipients with either anti-helper T-cell antibodies, or fusion proteins which block costimulatory interactions between antigen presenting cells and host T-cells have demonstrated synergy in significant prolongation of encapsulated islet xenograft survival in NOD mice with spontaneous diabetes. Technical improvements in microcapsule design also have contributed to prolonged graft survival. "Double-wall" microencapsulation provides a more durable microcapsule and islet pretreatment prior to encapsulation reduces the frequency of defective capsules with islets entrapped in the membrane. Long-term durability of encapsulated islet grafts remains a concern and further improvements in microcapsule design are a prerequisite to clinical trials.

Parathyroid hormone content distinguishes true normal parathyroids from parathyroids of patients with primary hyperparathyroidism.

The purpose of this study was to clarify the pathophysiology of primary hyperparathyroidism by looking for differences between parathyroids from eucalcemic patients and patients with primary hyperparathyroidism (HPT) with respect to the following parameters: intracellular parathyroid content of parathyroid hormone (PTH) and parathyroid hormone messenger RNA (PTH mRNA); and serum PTH and calcium levels of patients and patient age. Coded samples of human parathyroid biopsies were assayed for PTH content with a C-terminal-specific radioimmunoassay. Total cellular RNA was extracted, and PTH mRNA was quantified by dot-blot analysis. These results were tabulated along with associated data on patient age, preoperative serum PTH, and preoperative calcium levels. The content of PTH was significantly higher in true normal (TN) parathyroids than in parathyroids from patients with hyperparathyroidism. PTH content of adenomas and hyperplasias were similar. PTH content of normal parathyroids biopsied from patients with parathyroid adenomas (NA) was statistically higher than that of adenomas but statistically lower than that of TN parathyroids. PTH mRNA and PTH content were correlated (p < 0.001) for TN parathyroid glands; however, it was not true for glands (grossly normal or otherwise) in patients with HPT. Patient groups were similar with regard to mean patient age and intracellular PTH mRNA levels. Hypercalcemic patients were similar with regard to preoperative serum calcium and PTH levels. NA parathyroids, adenomas, and hyperplasias are different from TN parathyroids with regard to their PTH content. PTH mRNA was similar across all groups. The relation between intracellular PTH mRNA and PTH was significantly absent in patients with HPT compared with TN glands. Furthermore, we have found that PTH content of normal parathyroid in patients with adenoma is similar to that of hyperplastic and adenoma tissues. These data suggest that the PTH content of parathyroid tissues may be of use in differentiating normal from abnormal parathyroids.

Effects of neonatally administered iprindole on adult behaviors of rats.

In past studies, administration of the antidepressant drugs clorimipramine, zimeldine, or desipramine to neonatal rats produced abnormalities in adult rats that modeled some behavioral and/or REM sleep features of human endogenous depression. Although these three drugs affected different neurotransmitter systems, all caused REM sleep deprivation (RSD). This suggested the hypothesis that RSD of neonatal rats caused their adult depression. One prediction of this hypothesis is that neonatally administered iprindole, an antidepressant drug that does not produce RSD, will not produce adult rats that model depression. The present study tested this hypothesis. Iprindole was administered to neonatal experimental rats and saline was administered to neonatal control rats. When the rats matured, compared with control rats, experimental rats were not significantly different in aggressive behavior (shock induced fighting), sexual behaviors, open field locomotion, and REM sleep. In our previous studies on rats, all these adult behaviors were affected in a depressive-like way by neonatally administered clorimipramine. Because iprindole does not decrease REM sleep, the present results support the hypothesis that in rats neonatal RSD causes adult depression.

Dose-dependent decrements in adult male rat sexual behavior after neonatal clorimipramine treatment.

Previous studies found that months after clorimipramine (CLI) treatment of male neonatal rats, the mature animals developed behavioral deficits and REM sleep abnormalities that modeled human endogenous depression. In the initial studies neonatal rats received CLI 30 mg/kg/IP daily from age 8 through 21 days. Diminished sexual activity of the adult rats treated neonatally in this manner was a behavioral deficit that supported the depression model. However, in subsequent studies in our laboratory, the same neonatal treatment occasionally failed to produce adult sexual deficiencies found in the initial studies. The inconsistency raised the possibility that neonatal CLI treatment was not a reliable method to produce an animal model of depression. An alternative hypothesis was that the CLI dose was too low. The present study tested this hypothesis. Placebo or one of four CLI doses was administered daily to male neonatal rats (n = 12/treatment group) from age 8 days through 21 days: 30 (the original dose), 40, 50, and 60 mg/kg/IP. Six components of adult sexual behavior were measured at age 5 months. Deficiency of each sexual behavior was found to be dose-dependent (r = 0.5, p < 0.001). The 30 mg/kg/day dose caused deficiencies in some, but not all, sexual behavior measures. Higher doses caused deficiencies in all measures of sexual behavior. The results support the hypothesis that neonatal CLI treatment at doses higher than the original 30 mg/kg/day caused reliable impairments in adult male rat sexual behavior, and therby support the reliability of neonatal CLI treatment to produce an animal model of endogenous depression.

DNA index and ploidy distinguish normal human parathyroids from parathyroid adenomas and primary hyperplastic parathyroids.

BACKGROUND: The goal of this study was to identify factors that might aid in diagnosis and intraoperative management of hyperparathyroidism. METHODS: We analyzed biopsy specimens of 242 parathyroids from 159 patients by use of flow cytometry and image cytometry (ICM) for DNA index (DI), defined as the content of nuclear DNA compared with that expected for a DNA diploid standard, for proliferative index (PI), and for ploidy (diploid versus aneuploid or tetraploid). RESULTS: True normal and normal parathyroids from patients with solitary adenomas were uniformly diploid. Abnormal ploidy (aneuploidy or tetraploidy) was identified frequently in adenomas and occasionally in hyperplasias with the exception that multiple endocrine neoplasia (MEN) biopsy specimens were uniformly diploid. DI for adenomas was similar to that for hyperplasias, and DI of both was higher than for normal glands. ICM-DI correlated positively with flow cytometry-DI and patient age and inversely with serum parathyroid hormone. PI was relatively low in all groups but was higher for hyperplasias versus normal parathyroids from patients with solitary adenomas and MEN versus non-MEN. PI correlated inversely with patient age. CONCLUSIONS: DI by ICM differentiates normal from abnormal parathyroids. DI might influence extent of resection in two- and three-gland hyperplasia and selection of the most appropriate gland for autografting and cryopreservation in patients with four-gland hyperplasia.

Auditory brain-stem responses in neonates receiving extracorporeal membrane oxygenation.

Auditory brain-stem responses from 25 neonates treated with extracorporeal membrane oxygenation were compared with those of 11 control subjects. Results revealed no statistically significant differences for recorded responses, either between ears or between groups. We conclude that infants who receive extracorporeal membrane oxygenation, with or without carotid artery repair, are not at greater risk for auditory brain-stem dysfunction than similar infants who do not receive extracorporeal membrane oxygenation.

Procedure- and age-dependent hyperactivity in a new animal model of endogenous depression.

We have replicated the findings of Mirmiran and colleagues that neonatal administration of the antidepressant clomipramine (CLI) to male rats results in hyperactivity in open-field tests in adulthood. We report that this effect does not reliably occur in a "Digiscan" activity device. The difference in effect between the two activity measuring devices may be due to more stress being present in the open-field test, and we propose that the CLI-treated rats may be more reactive to stress. This hypothesized enhanced reactivity to stress may be similar to the proposed vulnerability of depressed humans to stress. In addition, we have found that the open-field effect does not occur until the rats are at least 4 months old; this delayed effect may be analogous to the progressive onset of endogenous depression in humans.

Decreased intracranial self-stimulation in a new animal model of endogenous depression.

Neonatal treatment of rats with clomipramine may produce adult animals which model endogenous depression. We report here that a major factor of depression in humans, the diminished capacity for pleasure, appears present in these rats. At age 7 months, bar-press responding for rewarding hypothalamic stimulation is reduced across a range of intensities. At age 4 or 5 months this effect is not seen, although other behavioral abnormalities are present at the younger age. The delayed onset of diminished intracranial self-stimulation may relate to the gradual insidious onset of endogenous depression in humans.

Diminished sexual activity in a new animal model of endogenous depression.

Our laboratory has proposed a new animal model of endogenous depression. The proposal is that in rats neonatal clomipramine (CLI) produces adult animals that model endogenous depression. Diminished sexual activity is a salient behavioral abnormality found in endogenous depression. This suggests that an animal model of endogenous depression should show diminished sexual activity. We report here a test of the prediction that after neonatal treatment with CLI, adult male rats show decreased sexual activity. We found that after neonatal CLI, adult male Long-Evans rats had a pervasive diminution of sexual activities including decreased mounts, intromissions, ejaculations, and increased mount latencies and postejaculatory pause. Sprague-Dawley and Wistar strains also tended to show decreased intromissions and ejaculations, but their baseline sexual activity was too low to give interpretable data. The results with the sexually active Long-Evans strain are consistent with the hypothesis that neonatal CLI produces adult rats that model human endogenous depression.

REM sleep abnormalities in a new animal model of endogenous depression.

Endogenous depression has reliable REM sleep abnormalities. These include a short REM latency, frequent sleep onset REM periods, and after REM sleep deprivation (RSD), an abnormal temporal course of REM rebound in the presence of a normal total REM rebound. The reliability of these abnormalities suggests that they ought to be present in an animal model of endogenous depression. In 1982, we proposed a new animal model of endogenous depression. Our hypothesis is that in rats neonatal clomipramine (CLI) will produce adult animals that model endogenous depression. In this study we tested the prediction that after neonatal treatment with CLI, adult rats will show the above three REM sleep abnormalities of human endogenous depression. We found that neonatal treatment with CLI produced rats that at age 6 months had shorter REM latency, more sleep onset REM periods than control rats, and after RSD, had an abnormal temporal course of REM rebound in the presence of a normal total REM rebound. The finding of these REM sleep abnormalities supported the validity of the animal model of endogenous depression.

A new animal model of endogenous depression: a summary of present findings.

In 1982 our laboratory proposed a new animal model of endogenous depression. The proposal was that in rats, neonatally administered clomipramine (CLI) will produce adult animals that model endogenous depression. We summarize here several tests of the validity of the model. Results were that after neonatal CLI, adult male rats showed behavioral abnormalities of the human disorder: decreased sexual, aggressive, and intracranial self-stimulation activities, as well as motor hyperactivity in a stressful situation. Preliminary evidence suggested that behavioral abnormalities in rats (sexual, aggressive, and motor) briefly treated with antidepressant treatments (imipramine, REM sleep deprivation) begin to normalize. Lastly, after neonatal CLI, the adult rats showed REM sleep abnormalities of endogenous depression, viz, low REM latency, frequent sleep onset REM periods, and abnormal temporal course of REM rebound after REM sleep deprivation. These results supported the hypothesis that in rats neonatal CLI produced adult animals that modelled endogenous depression.

Animal depression model by neonatal clomipramine: reduction of shock induced aggression.

Clomipramine, administered to neonatal rats, has been reported to produce adult behavioral and REM sleep abnormalities. They include decreased sexual behavior, increased ambulation in the outer part of an open-field arena, increased REM sleep % of total sleep time, and in descriptive data, short REM latency, and increased REM phasic events. Since these abnormalities resemble some found in human endogenous depression, we have hypothesized that the adult rats represent an animal model of depression. Diminished aggressive behavior is a common characteristic of endogenous depression. This study tested the validity of the animal depression model by determining in rats the effect of neonatal clomipramine on adult shock-induced fighting. Experimental rats were treated neonatally with clomipramine and control rats were treated neonatally with saline. When they matured, compared with control rats, experimental rats had significantly fewer offensive fighting responses, and significantly more defensive fighting responses. The findings add some support to the validity of the animal depression model produced by neonatal clomipramine.

Correlation properties of random phase diffusers for multiplex holography.

Random phase diffusers used to individually code the reference beams employed in multiplexing a set of holograms are best characterized by their autocorrelation and cross-correlation properties. In this paper multilevel (n-level) phase diffusers and ground-glass diffuser models based on a spatial random telegraph wave are employed to investigate the performance of such diffusers for both plane wave and spherical wave illumination. The advantage of using balanced phase diffusers is indicated, and it is shown that a binary phase diffuser can, in principle, perform as well as a multilevel (n > 2) diffuser, and even as well as ground glass if the spatial fineness of the diffusers is comparable. Two signal-to-noise ratio measures of the performance of the various diffuser systems in a multiplex holography application are evaluated and discussed.

Multiplex hologram representations of space-variant optical systems using ground-glass encoded reference beams.

Multiplex holograms can accurately represent 2-D space-variant optical systems if hologram-to-hologram cross talk can be reduced to an acceptable level. In the experiments reported here the combination of ground-glass diffusers with chirped wave front illumination proved to be an effective means of suppressing this cross talk. The residual cross talk distribution and intensity depend both on the signal and the optical system. The experiments also show that overlapping outputs from multiplexed holograms add coherently in amplitude. In addition, an accurate holographic representation of an extremely space-variant system is presented.