Pharmacoresistant convulsions and visual hallucinations around two weeks after selegiline overdose: a case report.

A case of pharmacoresistant convulsions after selegiline overdose is reported. A 50-year-old male having been suffering from bipolar II disorder for 16 years attempted suicide by taking an overdose of 195 mg selegiline with other psychotropics. He developed recurrent pharmacoresistant seizure from 12th day to 19th day after selegiline overdose. He also had visual hallucinations and temporary high blood pressure. The authors suspect that the catecholamine-influenced convulsions and visual hallucinations that manifested during the period increased by the MAO-inhibiting action of selegiline which lasts about 2 weeks.

High-throughput applicable genomic sex typing of chicken by TaqMan real-time quantitative polymerase chain reaction.

Genetic sex typing of vertebrate animals is an essential technique for research on reproductive phenomena such as sex determination of embryonic tissues. Polymerase chain reaction amplification of genomic DNA segments in the Z and W sex chromosomes has been widely used as a standard laboratory method to determine genetic sex of the chicken (Gallus gallus domesticus). However, conventional protocols for PCR determination of avian sex typically involve tedious steps of genomic DNA isolation, which often require relatively large amounts of tissue samples, and the purity of genomic DNA specimens significantly affects PCR efficiency. Moreover, detection of sex chromosome-specific PCR products by gel electrophoresis is prone to misjudgment caused by amplification of contaminating genomic DNA segments derived from tissue or DNA samples as well as previously generated PCR products. Thus, the credibility of genetic sex typing by conventional PCR-based methods that measure the relative amounts of the end product DNA amplicons critically depends on several experimental steps that are potentially vulnerable to errors. Here, we describe an optimized protocol of chicken genetic sex typing by TaqMan real-time quantitative PCR amplification of markers on the sex chromosomes. This TaqMan sex typing method accurately quantifies relative amounts of the Z and W sex chromosome markers directly from only 0.5 to 2 microL of total blood lysate without nucleic acid purification. The real-time amplification curves of the quantitative PCR reaction readily distinguished truly homozygous (ZZ) and heterozygous (ZW) sex chromosomes from contamination of the sex chromosomal DNA, ensuring highly credible sex determination. Thus, the TaqMan typing of chicken genetic sex has several advantageous features for high-throughput operation compared with conventional methods.

Nodes-and-connections RNAi knockdown screening: identification of a signaling molecule network involved in fulvestrant action and breast cancer prognosis.

Although RNA interference (RNAi) knockdown screening of cancer cell cultures is an effective approach to predict drug targets or therapeutic/prognostic biomarkers, interactions among identified targets often remain obscure. Here, we introduce the nodes-and-connections RNAi knockdown screening that generates a map of target interactions through systematic iterations of in silico prediction of targets and their experimental validation. An initial RNAi knockdown screening of MCF-7 human breast cancer cells targeting 6560 proteins identified four signaling molecules required for their fulvestrant-induced apoptosis. Signaling molecules physically or functionally interacting with these four primary node targets were computationally predicted and experimentally validated, resulting in identification of four second-generation nodes. Three rounds of further iterations of the prediction-validation cycle generated third, fourth and fifth generation of nodes, completing a 19-node interaction map that contained three predicted nodes but without experimental validation because of technical limitations. The interaction map involved all three members of the death-associated protein kinases (DAPKs) as well as their upstream and downstream signaling molecules (calmodulins and myosin light chain kinases), suggesting that DAPKs play critical roles in the cytocidal action of fulvestrant. The in silico Kaplan-Meier analysis of previously reported human breast cancer cohorts demonstrated significant prognostic predictive power for five of the experimentally validated nodes and for three of the prediction-only nodes. Immunohistochemical studies on the expression of 10 nodal proteins in human breast cancer tissues not only supported their prognostic prediction power but also provided statistically significant evidence of their synchronized expression, implying functional interactions among these nodal proteins. Thus, the Nodes-and-Connections approach to RNAi knockdown screening yields biologically meaningful outcomes by taking advantage of the existing knowledge of the physical and functional interactions between the predicted target genes. The resulting interaction maps provide useful information on signaling pathways cooperatively involved in clinically important features of the malignant cells, such as drug resistance.

Oxidative stress and disturbed glutamate transport in hereditary nucleotide repair disorders.

Xeroderma pigmentosum group A (XPA) and Cockayne syndrome (CS) are hereditary DNA repair disorders complicated by progressive neurodegeneration. Here we immunohistochemically examine the in situ expression of materials that are produced by oxidative stress and glutamate transporters (which can contribute to prevention of glutamate neurotoxicity) in the brains of 5 autopsied patients each of XPA, CS, and control groups. All oxidative products, including nitrotyrosine, advanced glycation end product, and 4-hydroxy-2-nonenal-modified protein (HNE) were deposited in large amounts in the globus pallidus of CS patients compared to XPA patients. They were frequently recognized in the pseudocalcified foci and free minerals in the neuropil, and more rarely in foamy spheroids. In addition, the deposition of HNE was observed also in hippocampal and cerebellar dentate neurons of both CS and XPA patients. The expression of glial glutamate transporters, EAAT1 and GLT-1, was affected in the globus pallidus in 5 CS patients and 3 XPA patients. They were also altered in the cerebellar cortex in most of the CS patients. These data suggest that oxidative stress and disturbed glutamate transport may be involved in pallidal and/or cerebellar degeneration in hereditary nucleotide repair disorders.

Cerebellar neurodegeneration in human hereditary DNA repair disorders.

Recent findings have focused attention on the role of apoptosis in neurodegenerative diseases, however, the apoptotic process in child-onset brain disorders has been little investigated. Xeroderma pigmentosum (XP) and Cockayne syndrome (CS) are hereditary disorders characterized by impaired DNA repair and neurodegeneration. We investigated apoptotic cell death in the cerebellum of five cases of XP group A (XPA), four cases of CS, and twelve controls, using TdT-mediated DIG-dUTP nick-end labeling (TUNEL) and immunohistochemical staining for bcl-2, bcl-x, p53, bax, BDNF and Trk B. The TUNEL-positive cells were found in the granule cells of the cerebellar cortex of two patients with XPA and two patients with CS, whereas such cells were not detected in the cerebellar cortex in controls. Upregulation of bcl-2 or BDNF was not observed, and bcl-x expression was not altered. Some patients showed nuclear expression of p53 in the granule cells and/or molecular layer, bax-positive glial cells in the cerebellar white matter, and a few Trk B-positive cells in the granular layer. These findings suggest that apoptotic cell death can be involved in the cerebellar degeneration in patients with hereditary defects in DNA repair mechanisms.

Neurodegeneration in hereditary nucleotide repair disorders.

Both xeroderma pigmentosum group A (XPA) and Cockayne syndrome (CS) are rare autosomal disorders, have a genetic defect in the step of nucleotide repair, and involve various neurological abnormalities caused by progressive neurodegeneration. We performed comprehensive neuropathological analysis of five cases of XPA and four cases of CS. The XPA cases showed widespread neuronal loss throughout the central nervous system, in sharp contrast to the comparative preservation of neurons in the CS cases, who rather exhibited patchy demyelination in the cerebral and cerebellar white matter, and multifocal calcium deposition in the basal ganglia and cerebral white matter, respectively. Exceptionally in the cerebellar cortex, neuronal loss was more severe in CS than in XPA. Grumose or foamy spheroid bodies occurred in the globus pallidus and substantia nigra, and axonal torpedoes were increased in the cerebellar cortex in both disorders. Neither silver impregnation nor immunohistochemistry for ubiquitin or tau succeeded in visualizing neurofibrillary tangles, senile plaques or augmented ubiquitination in either disorder, and these findings did not support the involvement of facilitated aging in the neurodegeneration in XPA or CS.

Membranous lipodystrophy (Nasu disease): clinical and neuropathological study of a case.

We report a sibling case of Nasu disease. A 35-year-old housewife, whose parents were consanguineous and whose sister died of the same disease, developed dementia, followed by bone fracture, incontinence and convulsions. She died at age 41. Pathologically, characteristic membranocystic changes of the fat cells in bone marrow and adipose tissues were observed. Neuropathologically, there was demyelination associated with intense gliosis and numerous axonal spheroids in the cerebral white matter. At the electron microscope level, these spheroids were an accumulation of various cell organelles. In addition, some had Hirano bodies. Incontinence was correlated with reduction of the number of nerve cells in Onuf's nucleus of the sacral cord.

[Neuropathological study of severely handicapped children: relationships of cortical and subcortical destructive lesions].

Neuropathological examinations were performed on 30 autopsy cases of severely handicapped children. Among them, 11 cases showed bilateral cerebral destructive lesions. The cerebral lesions were divided into three groups; six cases with dominantly grey matter lesions, three with dominantly white matter lesions and two with combined grey and white matter lesions. The cortical lesions were found in the fronto-parieto-occipital lobes and cingulate gyri, while undersurface of the temporal lobes showed less destruction. The white matter lesions, consisting of marked gliosis and atrophy accompanied by ventricular dilatation, were remarkable in the area extending from the periventricular region to the centrum semiovale. These changes were more apparent in the occipital lobes. Cerebellar lesions were found in nine cases, which also were classified into grey matter and white matter lesions. The extent and characteristics of these lesions resembled those of the cerebral lesions. The basal ganglia showed no remarkable destruction in the cases with severe cortical and subcortical damages. It was assumed that these nuclei had survived the disconnection from the cortex. Thalamic lesions were observed in six cases, mainly restricted to the dorsal and/or lateral nuclei. No relationship was found between these thalamic lesions and the extent or intensity of cerebral destruction. It is suggested that each of the thalamic nuclei has a different characteristic vulnerability to such destructive conditions.

[A case of Klippel-Trenaunay-Weber syndrome accompanied by congenital hydrocephalus and micropolygyria].

We report a patient with the Klippel-Trenaunay-Weber syndrome accompanied by congenital hydrocephalus, which was slowly progressive and an Ommaya's reservoir was set up. However, the hydrocephalus remained stable even when the shunt was removed due to infection. Generalized tonic clonic convulsions had appeared from six months after birth and were treated with valproic acid. The electroencephalogram showed hypsarrhythmia. He died at eight months of age. Autopsy revealed extensive micropolygyria of the bilateral cerebral hemispheres and hydrocephalus. To our knowledge, it is rare for the Klippel-Trenaunay-Weber syndrome to be accompanied by congenital hydrocephalus, and there has been no previous report of its occurrence with micropolygyria.

[An autopsy case of Down syndrome in an adult patient with multiple senile plaques and systemic amyloidosis].

A rare adult autopsy case of Down syndrome was reported. The patient was a 36-year-old male, whose chromosome analysis revealed 47, XY, +21. He showed typical systemic AA-amyloidosis and numerous senile plaques in the brain. Senile plaques were diffuse or primitive. They were composed of beta-protein, but negative for Congo-red stain. There were few neurofibrillary changes in the para hippocampal gyri. Nucleus basalis Meynert showed no significant lesion. The distribution of these plaques had some characteristics different from that for Alzheimer's disease. In the brain involvement of systemic AA-amyloidosis was restricted to the regions devoid of blood-brain-barrier, such as choroid plexus and pituitary gland. Cerebral beta-amyloid and systemic amyloid A protein were segregated on each side of the blood-brain-barrier. Therefore, we suggested that each amyloid might be synthesized and deposited by its own mechanism. Electronmicroscopically Hirano's body was identified in the hippocampal gyri.

[Female autopsy case of Seitelberger's connatal form of Pelizaeus-Merzbacher disease].

We report here an autopsy case of connatal Pelizaeus-Merzbacher disease, the second from Japan. Her clinico-pathological findings were essentially the same as those of the first case that we reported previously. The clinical course of this patient was 19 years in duration. Pathologically myelin had disappeared from the entire central nervous system, whereas that of the peripheral nervous system was preserved. Axons appeared also intact except for torpedo formation in the cerebellum. Demyelinated areas showed isomorphic gliosis. Recent studies have revealed impairment of proteolipid protein synthesis in classical Pelizaeus-Merzbacher disease, the causative gene of which being located on the X chromosome. Thus, this disease is inherited as an X-linked recessive trait. However we report herein a sporadic female case in a non-consanguineous family. Therefore, we propose that this disease might have another causative gene and/or another mode of inheritance.

[A case of myeloma kidney complicated by extramedullary plasmacytoma with massive bleeding].

A 76-year-old woman, who had received hemodialysis due to chronic renal failure of unknown cause for two months, was admitted to our hospital. She was suffering from severe pain in the left thigh, rapidly progressive anemia and thrombocytopenia after receiving a contusion on her left thigh. Soon after admission, the patient died of shock. Autopsy revealed multiple myeloma(lamda type) with extramedullary plasmacytoma and systemic amyloidosis. In the kidney, there were typical tubular casts with multinucleated giant cells and interstitial fibrosis. More specific findings included an extramedullary plasmacytoma in the left iliopsoas muscle surrounded by a huge hematoma. Internal hemorrhage resulting from indirect contusion at this site was likely to have caused her shock. Since typical clinical findings of multiple myeloma, such as serum M protein and hypercalcemia, were not found in this case, it was difficult to make a diagnosis of multiple myeloma. In case of multiple myeloma, micro- or macroscopic extramedullary tumor formation is not rare, but there has been no report of a case with macroscopic tumor formed in skeletal muscle, exhibiting massive hemorrhage. We report here a case of multiple myeloma with an unusual clinical course.

[Yolk sac tumor resected by transverse sternotomy: a case report].

Transverse sternotomy was performed in a 39-year-old man having bilateral lung metastatic lesions of yolk sac tumor. This approach provided an excellent operative field. Bilateral partial lobes including parietal pleura and diaphragma were readily resected. As described in this paper, transverse sternotomy is one of the beneficial method in the thorough surgical treatment for metastatic bilateral lung lesion.