Periventricular leucomalacia (PVL)-like lesions in two neonatal cynomolgus monkeys (Macaca fascicularis).

Periventricular leucomalacia (PVL) is a lesion of immature cerebral white matter that occurs in the perinatal period. In man, PVL is the predominant form of brain injury and a cause of cerebral palsy and cognitive deficits in premature infants. PVL affects fetuses and newborns, particularly those who have undergone oxygen deprivation as may occur in premature birth. Many clinical and pathological studies of PVL have been performed in man, but there is no clear definition of PVL in animals. A few spontaneous PVL-like cases in puppies or experimental cases in other animal species have been reported. The present study reports the histopathological and immunohistochemical features of PVL-like lesions in two neonatal cynomolgus monkeys. In both cases, there was cerebral white matter necrosis with marked infiltration of lipid-laden phagocytes and a reduction of neurons in the cerebral cortex. In case 1 there was extensive cavitation of the cerebral white matter. In case 2 there was reactive astrocytosis associated with a decrease in oligodendroglial cells and a decrease in cerebral white matter myelin. To our knowledge, this is the first report of PVL-like leucoencephalomalacia in non-human primates.

Acute megakaryocytic leukaemia (AMKL)-like disease in a cynomolgus monkey (Macaca fascicularis).

A 5-year-old male cynomolgus monkey (Macaca fascicularis) with a clinical history of bleeding tendency, severe anaemia, thrombocytopenia and elevated serum concentration of liver-related enzymes was examined post mortem. Ecchymotic haemorrhages were present on the left eyelid and forehead. The liver, kidney and spleen were markedly enlarged and the kidneys had capsular petechiae. Microscopically, numerous atypical cells resembling myeloid cells were observed in the bone marrow, and myelofibrosis was present. Atypical cells were also present in the blood vessels of the liver, kidney, spleen, lymph nodes, lung, heart, bladder, adrenal gland and brain. Some neoplastic cells had oval or pleomorphic macronuclei and others were multinucleated. Immunohistochemically, the majority of the neoplastic cells had granular cytoplasmic expression of the megakaryocyte-associated antigens Von Willebrand Factor and CD61-IIIa, but were negative for myeloperoxidase. A diagnosis of acute megakaryocytic leukaemia (AMKL)-like disease was made. This would appear to be the first report of AMKL-like disease in non-human primates. This monkey was infected with simian retrovirus type D and it is possible that this viral infection was associated with the development of neoplasia.

Congenital cystic adenomatoid-like malformation in a cynomolgus monkey (Macaca fascicularis).

Congenital cystic adenomatoid malformation (CCAM) is a developmental lung abnormality characterized by abnormal proliferation of mesenchymal elements and failure of bronchiolar structures to mature, ultimately resulting in the compression of normal pulmonary tissue and mediastinal shift with rapid expansion of cysts. Although various clinical and pathologic studies of CCAM in humans exist, CCAM has yet to be reported in animals, even in nonhuman primates. In the present study, histopathologic analyses of a neonatal cynomolgus monkey that died 17 days after birth revealed that normal lung architecture was replaced by disorganized overgrowths of cysts lined with simple cuboidal epithelium. The epithelium projected a few ciliates into the air spaces and produced mucus. To our knowledge, this is the first case study describing CCAM or a CCAM-like lesion in nonhuman primates.

Chloroplastic ascorbate peroxidase is the primary target of methylviologen-induced photooxidative stress in spinach leaves: its relevance to monodehydroascorbate radical detected with in vivo ESR.

Methylviologen (MV) induces oxidative damages in leaves. In order to understand its mechanism we studied initial biochemical events under light in MV-fed spinach leaves. When isolated chloroplasts were illuminated in the presence of MV, both stromal and thylakoid-bound ascorbate peroxidases (APX) were inactivated rapidly at the same rates, and their inactivation was retarded by ascorbate (AsA) at higher concentrations. Since MV accelerates the photoproduction of O2- in Photosystem (PS) I and simultaneously inhibits the photoreduction of monodehydroascorbate (MDA) to AsA, the inactivation of APX was attributed to the loss of AsA and accumulation of H2O2 in the stroma. Following APX, superoxide dismutase and NADP(+)-glyceraldehyde 3-phosphate dehydrogenase, both of which are vulnerable to H2O2, were inactivated by MV plus light. Dehydroascorbate reductase, monodehydroascorbate reductase, PS II, PS I and ferredoxin-NADP(+) reductase were far less sensitive to the treatment. In the treated leaves, cytosolic APX and guaiacol-specific peroxidase were also inactivated, but slower than chloroplastic APXs were. Catalase was not inactivated. Thus the MV-induced photooxidative damages of leaves are initiated with the inactivation of chloroplastic APXs and develop via the inactivation of other H2O2-sensitive targets. The decay half-life of the MDA signal after a short illumination in the leaves, as determined by in vivo electron spin resonance spectrometry (ESR), was prolonged when the H2O2-scavenging capacity of the leaf cells was abolished by the inactivation of chloroplastic and cytosolic APXs. The measurement of MDA in leaves by ESR, therefore, allows to estimate in vivo cellular capacity to scavenge the photoproduced H2O2.

Characteristics of chemiluminescence observed in the horseradish peroxidase-hydrogen peroxide-tyrosine system.

Electrolysis or horseradish peroxidase (HRP)-catalyzed oxidation of tyrosine and bityrosine in aqueous solution at pH 7.4 resulted in light emission in the visible region. Electrolysis of tyrosine emitted light which peaked at 490 nm and was almost completely quenched by superoxide dismutase (SOD), while emission by bityrosine peaked at 530 nm. In the HRP-H(2)O(2)-tyrosine system the oxidation-reduction of tyrosine emitted light with two prominent peaks, 490 and 530 nm, and was not quenched by SOD. The phenoxyl neutral radical of the tyrosine in HRP-H(2)O(2)-tyrosine system was detected by electron spin resonance (ESR) spectrometry using tert-nitrosobutane as a spin trap; the spin adduct was found to adhere to the HRP molecule during the enzymatic reaction. Further, bityrosine was detected in the HRP-H(2)O(2)-tyrosine reaction system. Changes in absorption spectra of HRP and chemiluminescence intensities during HRP-catalyzed oxidation of tyrosine suggest that for photon emission compound III is a candidate superoxide donor to the phenoxyl cation radical of tyrosine on the enzyme molecule. The luminescence observed in this study might be originated from at least two exciplexes involved with the tyrosine cation radical (Tyr(*+)) and the bityrosine cation radical (BT(*+))

New retinoid X receptor subtypes in zebra fish (Danio rerio) differentially modulate transcription and do not bind 9-cis retinoic acid.

Retinoid X receptors (RXRs), along with retinoic acid (RA) receptors (RARs), mediate the effects of RA on gene expression. Three subtypes of RXRs (alpha, beta, and gamma) which bind to and are activated by the 9-cis stereoisomer of RA have been characterized. They activate gene transcription by binding to specific sites on DNA as homodimers or as heterodimers with RARs and other related nuclear receptors, including the vitamin D receptor, thyroid hormone receptors (TRs), and peroxisome proliferator-activated receptors. Two additional RXR subtypes (delta and epsilon) isolated from zebra fish cDNA libraries are described here; although both subtypes form DNA-binding heterodimers with RARs and TR, neither binds 9-cis RA, and both are transcriptionally inactive on RXR response elements. In cotransfection studies with TR, the delta subtype was found to function in a dominant negative manner, while the epsilon subtype had a slight stimulatory effect on thyroid hormone (T3)-dependent transcriptional activity. The discovery of these two novel receptors in zebra fish expands the functional repertoire of RXRs to include ligand-independent and dominant negative modulation of type II receptor function.

[Quality assurance of pathology laboratories: analysis of histopathological diagnosis of endoscopical biopsies from lower alimentary tract based on specimen types].

As a part of the internal quality assurance in the pathology laboratories, we focused on the diagnosis of biopsies from the lower alimentary tract, and investigated the number of independent lesions contributing the specimens, biopsy pieces, and paraffin blocks prepared in each pathological examination, from the standpoint of the types of biopsy procedures employed. In addition, we compared those of pre- and post-operative diagnoses, and examined the adequacy in the cases with repeated biopsies. Regarding the procedures, polypectomy and EMR (endoscopical mucosal resection) accounted for more than 60% of all the specimens handled during the study period. Most of the specimens taken by polypectomy or EMR were derived from neoplastic conditions, and roughly 40% of them were from malignancies including borderline lesions in a narrow sense. The examination of the surgically resected specimens confirmed a hundred percent accuracy ratio of the diagnosis on the biopsy specimens. Three cases (7.1%) out of 42 malignancies resected needed repeated biopsies in preoperative confirmation of malignancy, while the remainder (92.9%) had been diagnosed at the initial biopsy. The result disclosed that biopsies possessing both diagnostic and therapeutic value (polypectomy and EMR) are apparently increased in the field of lower alimentary tract, and the difficulty and efforts required for diagnosis are quite different and depend on the biopsy procedures employed.

The Drosophila nuclear receptors FTZ-F1 alpha and FTZ-F1 beta compete as monomers for binding to a site in the fushi tarazu gene.

The striped pattern of fushi tarazu (ftz) expression found in the blastoderm of the Drosophila melanogaster embryo is generated largely through complex interactions between multiple transcription factors that bind to the zebra element of the ftz gene. A motif in the zebra element, the FTZ-F1 recognition element (F1RE), has been shown to bind a transcription factor, FTZ-F1 alpha, that is a member of the nuclear receptor family. We recently identified a second, related member of this family, FTZ-F1 beta, that also binds to this motif. To investigate the possibility that FTZ-F1 alpha and FTZ-F1 beta coregulate ftz transcription through the F1RE, we have studied the DNA binding properties of FTZ-F1 alpha and FTZ-F1 beta. We demonstrate that recombinant FTZ-F1 alpha and FTZ-F1 beta proteins produce similar in vitro DNase I footprint patterns on a 14-nucleotide region of the zebra element and bind to this site with similar affinities and sequence specificities. Using wild-type and N-terminally truncated receptors, we have determined that FTZ-F1 alpha and FTZ-F1 beta both bind as monomers to the 9-bp F1RE in the zebra element, as well as to an imperfect inverted F1RE repeat present in the Drosophila alcohol dehydrogenase gene. A polyclonal antibody raised against FTZ-F1 beta identifies a predominant F1RE-binding component in embryonic nuclear extracts. Although FTZ-F1 alpha is also present in these extracts, FTZ-F1 alpha and FTZ-F1 beta do not appear to form heterodimers with each other. Cotransfection assays in mammalian cell culture indicate that both receptors contribute to the net transcriptional activity of a reporter gene through their direct interaction with the F1RE. These data suggest that FTZ-F1 alpha and FTZ-F1 beta likely coregulate common target genes by competition for binding to a 9-bp recognition element.

FTZ-F1 beta, a novel member of the Drosophila nuclear receptor family.

The Drosophila melanogaster gene FTZ-F1 beta, encoding a novel member of the steroid/thyroid hormone receptor gene superfamily, was isolated by cross-hybridization with a complementary DNA for the Drosophila nuclear receptor, FTZ-F1 (Lavorgna et al., 1991). The cDNA deduced protein sequence for FTZ-F1 beta displays significant amino acid identity with other vertebrate and invertebrate nuclear receptors, most notably with FTZ-F1. Also, bacterially expressed FTZ-F1 beta protein binds to a FTZ-F1 binding site found in the zebra stripe promoter element of the segmentation gene fushi tarazu (ftz). Northern blot analysis detected FTZ-F1 beta expression at all stages of the Drosophila life cycle including a possible maternal component. In situ hybridization in whole-mounted embryos localized transcripts for FTZ-F1 beta evenly expressed throughout the blastodermal layer in early embryos. At later stages of development strong FTZ-F1 beta expression is observed in both the brain and ventral chord structures as well as in the hindgut. Temporal and spatial expression patterns of the FTZ-F1 beta gene suggest that it may have multiple roles in early embryogenesis, neurogenesis, and in the adult. Furthermore, the identification of FTZ-F1 beta as a nuclear receptor family member suggests that an as yet undiscovered FTZ-F1 beta specific ligand is involved in Drosophila development.