A study of CR-39 plastic charged-particle detector replacement by consumer imaging sensors.

Consumer imaging sensors (CIS) are examined for real-time charged-particle detection and CR-39 plastic detector replacement. Removing cover glass from CIS is hard if not impossible, in particular for the latest inexpensive webcam models. We show that $10-class CIS are sensitive to MeV and higher energy protons and α-particles by using a 90Sr ß-source with its cover glass in place. Indirect, real-time, high-resolution detection is also feasible when combining CIS with a ZnS:Ag phosphor screen and optics. Noise reduction in CIS is nevertheless important for the indirect approach.

Qualitative comparison of bremsstrahlung X-rays and 800 MeV protons for tomography of urania fuel pellets.

We present an assessment of x-rays and proton tomography as tools for studying the time dependence of the development of damage in fuel rods. We also show data taken with existing facilities at Los Alamos National Laboratory that support this assessment. Data on surrogate fuel rods have been taken using the 800 MeV proton radiography (pRad) facility at the Los Alamos Neutron Science Center (LANSCE), and with a 450 keV bremsstrahlung X-ray tomography facility. The proton radiography pRad facility at LANSCE can provide good position resolution (<70 µm has been demonstrate, 20 µm seems feasible with minor changes) for tomography on activated fuel rods. Bremsstrahlung x-rays may be able to provide better than 100 µm resolution but further development of sources, collimation, and detectors is necessary for x-rays to deal with the background radiation for tomography of activated fuel rods.

The neutron imaging diagnostic at NIF (invited).

A neutron imaging diagnostic has recently been commissioned at the National Ignition Facility (NIF). This new system is an important diagnostic tool for inertial fusion studies at the NIF for measuring the size and shape of the burning DT plasma during the ignition stage of Inertial Confinement Fusion (ICF) implosions. The imaging technique utilizes a pinhole neutron aperture, placed between the neutron source and a neutron detector. The detection system measures the two dimensional distribution of neutrons passing through the pinhole. This diagnostic has been designed to collect two images at two times. The long flight path for this diagnostic, 28 m, results in a chromatic separation of the neutrons, allowing the independently timed images to measure the source distribution for two neutron energies. Typically the first image measures the distribution of the 14 MeV neutrons and the second image of the 6-12 MeV neutrons. The combination of these two images has provided data on the size and shape of the burning plasma within the compressed capsule, as well as a measure of the quantity and spatial distribution of the cold fuel surrounding this core.

Adhesion of polymorphonuclear leukocytes to protein-coated and platelet adherent surfaces.

Since protein adsorption and platelet adhesion are likely to precede significant contact of leukocytes with the surfaces of artificial organs, we have chosen to study polymorphonuclear leukocyte (PMN) adhesion in a sequential manner. The work presented here deals with the effects of flow and surface type on PMN adhesion to fibrinogen- and albumin-coated glass. We compared direct adhesion to adsorbed protein with adhesion to adsorbed protein having adherent platelets. These experiments were designed to see if PMN's might preferentially adhere to albumin or fibrinogen and whether a particular morphological form of adherent platelet could promote PMN adhesion. The adhesion of PMN's to spread platelets on albumin or fibrinogen occurs to a greater extent than in the absence of platelets. Adhesion of PMN's to spread platelets may be an important mechanism for their depletion from the circulation during artificial organ use.

Amylase expression in human parotid neoplasms: evidence by in situ hybridization for lack of transcription of the amylase gene.

Salivary alpha-amylase (EC 3.2.1.1) is the major protein component of human parotid gland secretion. We studied amylase gene structure and expression in tissue from a series of normal and neoplastic parotid glands by Southern blot analysis, in situ hybridization, and immunohistochemistry. Thirty-two tumors were examined. Southern blot analysis of DNA extracted from a Warthin tumor, an adenoid cystic carcinoma, and a mucoepidermoid carcinoma showed no evidence of structural rearrangement of amylase genes. Eleven parotid Warthin tumors were negative for amylase protein and mRNA by immunocytochemistry and in situ hybridization. One pleomorphic adenoma in the group of 10 examined showed focal staining for amylase protein, although amylase mRNA could not be demonstrated in the same population of cells by in situ hybridization in serial tissue sections. Five mucoepidermoid carcinomas and three acinar cell carcinomas were devoid of amylase protein and mRNA. Normal parotid tissue obtained from all patients studied revealed abundant acinar cell amylase mRNA and protein. In situ hybridization, in conjunction with immunocytochemistry, allows precise cellular localization of mRNA and protein, thereby establishing the site of production of specific transcripts. We conclude that the interruption in amylase gene expression in parotid gland neoplasms occurs at the transcriptional level.

Expression of human salivary protein genes.

Human proline-rich proteins (PRPs) are polymorphic, homologous in sequence, and linked in a cluster called the human salivary protein complex (SPC). Recently this complex was localized to human chromosome band 12p13.2 (Mamula et al., Cytogenet. Cell Genet. 39:279, 1985). We have isolated a PRP cDNA, EO27, from a human parotid gland library, identified it by DNA sequencing, and used it to study the molecular and cellular biology of PRP production. Cell-free translation and mRNA characterization with EO27 indicate that the numerous PRPs seen in saliva are produced from relatively few, large precursors, probably by posttranslational cleavage. This supports an hypothesis originally proposed by Friedman and Karn in 1977 (Am. J. Hum. Genet. 29:44 A; Biochem. Genet. 15:549) and later supported by biochemical studies (Karn et al., Biochem Genet. 17:1061, 1979) and molecular studies (Mamula et al., Fed. Proc. 43:1522, 1984; Maeda et al., J. Biol. Chem. 260:1123, 1985). EO27 was also used in this study to localize PRP mRNA production to the acinar cells of the parotid gland by in situ hybridization.

In situ localization of amylase mRNA and protein. An investigation of amylase gene activity in normal human parotid gland.

The distribution of human salivary amylase mRNA was studied by in situ hybridization to a [32P]-labeled amylase cDNA probe. Amylase mRNA was localized to the apical portion of acinar cells in frozen sections of human parotid salivary gland. No hybridization was noted in ductal cells, skeletal muscle, or in connective tissue. These results were consistent with immunohistochemical localization of amylase. The technique of in situ hybridization was modified to permit localization of amylase mRNA in variously fixed, paraffin-embedded parotid glands. Although the hybridization signal decreased with all fixatives, the pattern of localization paralleled that obtained with frozen sections. No advantage was noted in fixation with ethanol-acetic acid or Bouin solution over routine fixation with formalin. These results have important implications for researchers interested in studies of gene expression. We have demonstrated that routinely fixed paraffin blocks of human tissue can be used for cellular localization of specific mRNA. In coordination with immunocytochemistry, in situ hybridization offers a powerful tool for studies of mRNA and protein expression in individual cells.

Distribution of phosphodiesterase I in normal human tissues.

Phosphodiesterase I (PDE I) is an exonuclease capable of hydrolyzing a variety of phosphate ester and pyrophosphate bonds. Cell fractionation and histochemical studies in animal tissues have localized PDE I in the plasma membrane of various epithelia. This suggests a role for the enzyme in active transport. Distribution of PDE I in human tissues has not previously been studied. We have produced a polyclonal antiserum to bovine intestinal PDE I and have demonstrated crossreactivity with the human intestinal enzyme. This polyclonal antiserum was used in PAP immunocytochemistry to localize immunoreactive PDE I in a variety of human tissues. Localization was prominent in the gastrointestinal tract, including the cytoplasm of gastric mucosa parietal cells, cytoplasm of surface epithelium and isolated crypt cells in small intestine, and the colonic epithelial cytoplasm and brush border. Parotid gland acinar cells and scattered ductal cells showed positive cytoplasmic staining. Acinar and scattered pancreatic islet cells contained immunoreactive PDE I, as did Kupffer cells of the liver sinusoids. Immunoreactive PDE I was found in all vascular endothelia. The epithelium of the urinary tract showed extensive immunoreactivity. This included the distal convoluted and collecting tubules of the kidney, and ureteral and bladder urothelium. In previous histochemical studies of animal tissues, no evidence of PDE I activity was noted in male or female reproductive tract. In this study, immunoreactive PDE I was localized to human Sertoli cells and to basal epithelium of the epididymis and prostate acini. Fallopian tube epithelium of female reproductive tract also demonstrated immunoreactive PDI I, as did several cell types in term placenta. Our immunocytochemical results with human tissues differ significantly from previous histochemical studies in animal tissues, principally in the genitourinary system. This may be due in part to the different detection systems employed as well as the higher sensitivity of the immunoperoxidase technique. This underscores the importance of adjunct techniques in tissue surveys. The widespread epithelial distribution of immunoreactive PDE I detected by this polyclonal antibody implies an integral role in cell function, probably in active transport.

An amylase-producing serous cystadenocarcinoma of the ovary.

A patient with an amylase-producing serous cystadenocarcinoma of the ovary had elevated serum and urine amylase levels and high levels of amylase in pleural and ascitic fluids. Serum and urine amylase levels reflected both surgical removal of tumor mass and response to chemotherapy. Tumor homogenates had pronounced amylase activity. Salivary type amylase isozyme patterns were found in electrophoresis of samples from all sources. Ascites tumor cells were successfully cultured and salivary type amylase was found in the culture media throughout 5 passages. The tumor was classified by light microscopy as poorly differentiated serous cystadenocarcinoma. Ultrastructural studies on the tumor were consistent with that diagnosis. Amylase was detected in the cells of the tumor examined by the immunoperoxidase technique.

Immunohistochemical demonstration of ribonuclease and amylase in normal and neoplastic parotid glands.

Antibodies specific for bovine ribonuclease A (antiRNase A) were raised in rabbits, and immunologic cross-reactivity between bovine RNase A and human salivary gland RNase was demonstrated. The antiRNase A served as the primary antibody in the peroxidase-antiperoxidase immunohistochemical technique. Paraffin blocks of five normal human parotids and 20 parotid tumors were examined. In normal parotid and in cases of cystadenoma lymphomatosum, immunoreactive RNase was localized in the ductal epithelium, evidence of the ductal cell origin of these benign tumors. RNase immunoreactivity was noted in the adenomatous structures and in cells isolated in the myxoid matrix of pleomorphic adenomas, which supports recent evidence of an epithelial origin of these tumors. Malignant acinar cells of acinic cell carcinoma were strongly positive for immunoreactive RNase, while acinar cells of normal parotid were uniformly negative. This expression of the gene for RNase A probably represents a loss of differentiation (i.e., control) of the neoplastic acinar cells. Further evidence for this hypothesis was obtained by treating these tumors with an antihuman salivary amylase antibody, which is localized in normal acinar cells. No immunoreactive amylase was observed. The results support the idea that immunoreactivity need not accompany enzyme activity, as the presence of immunoreactive RNase was noted in all neoplastic tissues examined. Immunohistochemical localization of two antigens in the same tissue demonstrates the varied biochemical changes associated with parotid neoplasia.

Papillary serous carcinoma of the retroperitoneum.

We describe a retroperitoneal neoplasm in an 11-year-old girl which had a light microscopic appearance identical to that of papillary serous carcinoma of the ovary. There was no evidence of ovarian involvement. Immunohistochemical staining for amylase was positive within the cytoplasm of tumor cells. Since amylase is a marker for serous ovarian tumors, this finding supports the belief that "ovarian-type" neoplasms that occur at ectopic locations are essentially identical to their ovarian counterparts. We believe they originate from metaplasia of mesothelium. Our findings support the concept that these tumors should exhibit a biologic behavior and therapeutic response which are similar to those of an ovarian tumor of the same grade and comparable stage. The demonstration of intracytoplasmic amylase also may prove useful in differentiating peritoneal serous tumors from non-metaplastic mesothelial proliferations. We are unaware of a prior report of an extra-ovarian serous carcinoma in a child.

Hyperexplexia: an inherited disorder of the startle response.

A family is presented with hyperexplexia, a rare autosomal dominant neurological disorder. Affected individuals manifest flexor hypertonia and hypokinesia during infancy. Later and throughout life, the condition is characterized by exaggerated involuntary myoclonic startle reactions, which on occasion result in falling. There are also marked nocturnal myoclonic jerks. Many family members have had congenital hip dislocations and inguinal hernias. Pre- and postnatal hypertonia is proposed as the cause for these problems. The nature and location of central nervous system dysfunction in hyperexplexia was investigated using electroencephalographic and brainstem-evoked response techniques. A dysfunction of cortical inhibition of the brainstem-mediated startle response is discussed as a possible pathogenic mechanism. Accurate diagnosis of this disorder is important in order to provide appropriate counseling and to initiate effective treatment.

Fluorescent antibody study of the post-cysticercoid development of Moniezia expansa.

The strobila of Moniezia expansa was separated into developmental areas, and these were compared using immunological techniques. Agar double diffusion plates and immunoelectrophoresis showed differing antigenic composition or concentration between the strobilar regions studied. Conjugation of the antisera with rhodamine lissamine-200 aided in localization of common antigens on tissue sections of the various developmental stages. What appeared to be unique localizations were observed.