Determination of synapsin I and synaptophysin in body fluids by two-site enzyme-linked immunosorbent assays

Megalin, a 600 kDa membrane protein belonging to the IDL receptor family is highly expressed in the endocytic pathway of renal proximal tubules. In addition, this receptor is found in several other epithelia facing transcellular fluids but is also expressed in the parathyroid glands. Recent studies have established this protein as probably the most important receptor for endocytosis of macromolecules filtered in the renal glomeruli. The ligands reported to bind to megalin consist of a variety of different substances including albumin, vitamin-carrier complexes, proteinases and proteinase-inhibitor complexes, lipoprotein particles, receptor associated protein (RAP), different drugs and calcium.     

Inverse expression pattern of REST and synapsin I in human neuroblastoma cells

The zinc finger protein REST is a repressor of neuronal genes in nonneuronal tissues. We have analyzed the expression of REST, together with the expression of a REST target gene, encoding synapsin I, in human neuroblastoma cells. It was found that REST and synapsin I are coexpressed in neuroblastoma cell lines, although the expression of REST was inversely proportional to the levels of synapsin I mRNA. Thus, increased expression of synapsin I was directly correlated with decreased expression of REST. These expression data are in excellent correlation with synapsin I promoter activity measured in neuroblastoma cells showing that an increase in the REST concentration switched off synapsin I promoter activity. We conclude that the concentration of REST determines the expression level of neuronal genes such as the synapsin I gene.     

Differential ultrastructural distribution of synapsin and synaptophysin proximal to a ligation in bovine splenic nerve

Synaptophysin and synapsin, closely correlated on synaptic vesicles in terminals, may show a differential distribution at synapse formation and maturation. In order to disclose the fine structural details of these differences, synapsin and synaptophysin distribution was studied by immunocytochemistry on ligated bovine splenic axons in vitro and compared with terminals in the vas deferens. In the synaptic differentiations taking place proximally synapsin could only be detected on the accumulating elements of the axonal reticulum. Large dense granules and clusters of small synaptic vesicles were negative. Synaptophysin was restricted to these clusters. In the vas deferens, co-localization of synapsin and synaptophysin could be seen on small vesicles. From their formation small synaptic vesicles carry synaptophysin. Synapsin may be involved in the dynamic membrane changes taking place at the ligation. At a functional terminal, synapsin shifts to small synaptic vesicles.     

Comparative analysis of diagnostic techniques for localization of gastrointestinal neuroendocrine tumors

In vitro studies have shown that gastroenteropancreatic tumors, with the exception of insulinomas, have a high density of somatostatin receptors and can be imaged in vivo using somatostatin receptor scintigraphy (SRS) with either [123I-Tyr3]octreotide or [111In DTPA,DPhe1]octreotide. However, the sensitivity in relation to conventional imaging studies (ultrasound, CT, MRI, angiography) remains unclear. To address this question, we performed a prospective study of 80 patients with gastrinomas where SRS was compared with other conventional imaging techniques for detecting extrahepatic gastrinomas or liver metastases. Extrahepatic gastrinomas were identified by SRS in 58 percent of patients, whereas conventional imaging studies detected gastrinomas in 9 percent to 48 percent of patients. In detecting hepatic metastases in 24 patients with histologically-proven metastases, SRS was positive in 92 percent; ultrasound, CT or angiography in 42 percent to 62 percent; and MRI in 71 percent of patients. These results are compared with other studies in detecting gastrinomas as well as series including other PETs, excluding insulinomas, with insulinomas alone, and with carcinoid tumors. An analysis of the ability of SRS to identify gastrinomas found in different sites at surgery was performed. The role of endoscopic ultrasound (EUS) in detecting various PETs, in comparison to that of SRS, is yet to be established, particularly for extrapancreatic PETs. Therefore, the results of EUS in various studies containing patients with PETs are compared to those with SRS and conventional imaging studies. These data suggest that EUS is the first choice of localization methods for detecting insulinoma, which is an intrapancreatic tumor in almost all cases. In other PETs there still is not sufficient data to establish the relative roles of EUS and SRS.     

Cloning of human ENC-1 and evaluation of its expression and regulation in nervous system tumors

We recently identified and characterized a novel murine gene, ENC-1, that is expressed primarily in the nervous system and encodes an actin-binding protein. To gain insight into a potential role for ENC-1 gene in the processes of cell differentiation and malignant transformation in the human nervous system, we first cloned and characterized the human homologue of ENC-1. The human ENC-1 gene appeared to be highly expressed in adult brain and spinal cord, and in a number of cell lines derived from nervous system tumors we detected low steady-state levels of ENC-1 mRNA. We used a neuroblastoma differentiation model, the retinoic acid-induced neuronal differentiation of SMS-KCNR cells, to study the regulation of the ENC-1 gene during neural crest cell differentiation. We found that the expression of ENC-1 increased dramatically in the differentiated SMS-KCNR cells as compared to control undifferentiated cells. These results suggest that ENC-1 expression plays a role during differentiation of neural crest cells and may be down regulated in neuroblastoma tumors.     

Genetics of pediatric central nervous system tumors

PURPOSE: Pediatric central nervous system (CNS) tumors comprise a wide variety of histologic subtypes ranging from the benign juvenile pilocytic astrocytoma to the highly aggressive atypical teratoid/rhabdoid tumor. Although some brain tumors are seen in association with inherited genetic disorders which predispose to malignancies, most are sporadic. Current knowledge regarding the cytogenetic and molecular genetic events which have been implicated in the development or progression of common brain tumors in children in the subject of this review. METHODS: Combined cytogenetic and molecular genetic approaches, including fluorescence in situ hybridization, have been used to identify genomic alterations in different histologic types of pediatric brain tumors. RESULTS: The most frequent abnormality in primitive neuroectodermal tumor/medulloblastoma is an i(17q), present in approximately 50% of cases. This finding implicates the presence of a tumor suppressor gene on 17p, which is important in tumor development. A number of genes on 17p have been eliminated as candidates for this locus, including TP53. A tumor suppressor gene in chromosome band 22q11.2 has been hypothesized to play a role in atypical teratoid/rhabdoid tumors, and positional cloning strategies are in progress to identify a rhabdoid tumor gene. Chromosome 22 deletions are also seen in meningiomas and a small percentage of ependymomas, but it is not yet known whether the same gene is responsible for more than one malignancy. With regard to childhood astrocytomas, tumor-associated genetic changes have not yet been identified for the common juvenile pilocytic or low grade diffuse astrocytoma. In contrast, malignant anaplastic astrocytomas and glioblastoma multiforme have abnormalities similar to those seen in adults, including loss of alleles on 17p13 and TP53 mutations, trisomy 7, EGFR rearrangements, and loss of chromosomes 10 and 22. CONCLUSIONS: The presence of tumor-associated genetic abnormalities has clinical utility in a differential diagnostic setting, and has lead to the identification of genes which contribute to tumorigenesis.     

Gene therapy of central nervous system tumors

Recently, remarkable advances have been achieved in molecular and genetic researches of different kinds of general diseases, as well as in basic and clinical studies using gene therapy for central nervous system diseases. For brain tumors, clinical trials have been already started in more than 10 clinical protocols and more than 100 patients with malignant brain tumors. Nevertheless, there are still major issues that remain to be resolved for achieving better clinical results, such as delivery system of genetic material, regulatory methods of the intracellular expression of the transgene, antitumor efficacy, and tumor selectivity. In this paper, molecular genetic studies and the current state of gene therapy for neurological diseases, especially brain tumors, are described, and the future direction of this fascinating approach is discussed.     

Aggressive forms of gastric neuroendocrine tumors in multiple endocrine neoplasia type I

In recent classifications of gastric endocrine tumors, tumors arising in patients with multiple endocrine neoplasia type 1 (MEN-1) are regarded to be regulated by the concomitant hypergastrinemia resulting from to pancreatic or, most commonly, duodenal gastrinomas and to have a benign behavior. In this article, we report on two cases of MEN-1 gastric neuroendocrine tumors having a fatal course. Case 1 was a young male with hyperparathyroidism and Zollinger-Ellison syndrome and with florid development of multiple gastric carcinoids and multiple duodenal gastrinomas. Metastases occurred in the liver, of exclusive gastric origin, in periduodenal lymph nodes, of exclusive duodenal origin, and in perigastric lymph nodes, of mixed origin. The patient died 48 months after diagnosis. Case 2 was an adult female patient with hyperparathyroidism, adrenocortical disorders, and gastric tumors but no hypergastrinemia. The patient died 3 months after tumor diagnosis. At autopsy, the stomach showed multiple benign carcinoids and two independent neuroendocrine carcinomas not reported before in MEN-1 and massively metastatizing to lymph nodes, liver, and peritoneum. Multiple islet cell tumors mostly producing pancreatic polypeptide were found, whereas gastrinomas were seen in neither the pancreas nor the duodenum. Allelic losses at the MEN-1 gene locus in chromosome 11q13, the mechanism responsible for tumor development in MEN-1 syndrome, were demonstrated in the carcinoid tumors of case 1 and in the neuroendocrine carcinoma of case 2. We conclude that gastric neuroendocrine tumors in patients with MEN-1 may have a poor outcome, they have the same genetic mechanism as MEN-1 tumors in other organs, and they may be independent of the trophic effect of hypergastrinemia.     

Molecular and cell biological aspects of neuroendocrine tumors of the gastroenteropancreatic system

Neurons and neuroendocrine cells share a variety of common characteristics. Cell and molecular biological studies in recent years have improved our understanding of physiological and pathophysiological processes such as cellular growth, adhesion, and secretion of neuroendocrine cells. Here we review current findings from the area of basic research and from current clinical research relevant to improving the diagnosis and therapy of neuroendocrine tumors of the gastroenteropancreatic system.     

Recent advances in the biology of central nervous system tumors

BACKGROUND: In order to study the biosynthesis of vitamin B12, it is necessary to produce various intermediates along the biosynthetic pathway by enzymic methods. Recently, information on the organisation of the biosynthetic pathway has permitted the selection of the set of enzymes needed to biosynthesise any specific identified intermediate. The aim of the present work was to use recombinant enzymes in reconstituted multi-enzyme systems to biosynthesise particular intermediates. RESULTS: The products of the cobG and cobJ genes from Pseudomonas denitrificans were expressed heterologously in Escherichia coli to afford good levels of activity of the corresponding enzymes, CobG and CobJ. Aerobic incubation of precorrin-3A with the CobG enzyme alone yielded precorrin-3B. When CobJ and S-adenosyl-L-methionine were included in the incubation, the product was precorrin-4. Both precorrin 3B and precorrin-4 are known precursors of vitamin B12 and their availability has allowed new mechanistic studies of enzymic transformations. CONCLUSIONS: Our results show that the expression of the CobG and CobJ enzymes has been successful, thus facilitating the biosynthesis of two precursors of vitamin B12. This lays the foundation for the structure determination of CobG and CobJ as well as future enzymic experiments focusing on later steps of vitamin B12 biosynthesis.     

Primary central nervous system tumors: advances in knowledge and treatment

The ability to diagnose, monitor, and treat CNS tumors has been improved by new imaging techniques such as positron emission tomography (PET) scanning and functional MR imaging, stereotactic surgery, delivery of radiotherapy with brachytherapy and radiosurgery, and novel methods for delivering chemotherapy. These innovations combined with the new information about tumor pathogenesis and behavior revealed by molecular research give hope that more specific treatments for malignant CNS tumors will be developed in the future.     

Cytogenetic analysis of 109 pediatric central nervous system tumors

Reports of cytogenetic abnormalities in pediatric central nervous system (CNS) tumors are important for collection and comparison of large numbers of karyotypes of primary CNS neoplasms to produce statistically significant correlations. We report cytogenetic results of 119 samples of pediatric CNS tumors from 109 patients. Tumors included 33 low-grade astrocytomas, 18 high-grade astrocytomas, 14 gangliogliomas, 13 ependymomas, 17 primitive neuroectodermal tumors (PNET), three choroid plexus papillomas and carcinomas, and a miscellaneous group of 20 rare primary CNS tumors and metastases. In each group, cytogenetic results were correlated with histologic subtype and survival. The study indicated specific chromosome abnormalities in different groups of tumors. Low-grade astrocytomas showed mostly numeric abnormalities with gains of chromosome 7, high-grade astrocytomas showed differences from karyotypic changes observed in adults in lacking double minutes (dmin) and monosomy 10. The ependymoma group showed the largest proportion of abnormal karyotypes with frequent involvement of chromosome 6 and 16. Chromosome 6 was the single most common abnormal chromosome in this study, closely followed by chromosomes 1 and 11. Pediatric CNS neoplasms differ from adult tumors cytogenetically as well as histologically and biologically.     

Prognostic significance of ploidy level in human tumors. I. Carcinoma of the uterus

The 5-year survival rates of 540 patients with carcinoma of the cervix and 186 with carcinoma of the corpus uteri were assessed in relation to the modal DNA values of the tumors. Patients with squamous cell cervical carcinomas had more favorable prognoses if the modes were near-triploid or hypotetraploid; however, these high-ploidy tumors included more stage III cases than did the tumors with near-diploid modes. Patients with near-diploid endometrial carcinomas has considerably more favorable prognosis than did patients with the minority of tumors at this site, who had high modes; this prognostic difference was only partly related to a higher proportion of poorly differentiated tumors in the high-ploidy group since, among the poorly differentiated tumors, individuals with near-diploid modes again had significantly better prognoses than those with high modes.     

Induction of apoptosis in neuroendocrine tumors of the digestive system during treatment with somatostatin analogs

The frequency of antibiotic resistance among bacteria in 4 intensive care units (ICUs) at a university hospital in Sweden was investigated annually from 1993 to 1996. An increase in ampicillin-resistant enterococci from 1993 to 1995 was seen which was due to a shift from Enterococcus faecalis to Enterococcus faecium. After a special infection control programme was instituted, the rate of ampicillin resistance among enterococci and the number of E. faecium isolates declined during 1996. The oxacillin resistance rates for Staphylococcus aureus were < or = 2%, while most of the coagulase-negative staphylococci (CNS) were oxacillin resistant. No vancomycin-resistant enterococci or staphylococci were seen. The ciprofloxacin resistance rate for CNS and Enterococci spp. were high. Relatively, high levels of resistance to cefotaxime and piperacillin/tazobactam among Enterobacter spp. were also seen. During 1995 and 1996 Pseudomonas aeruginosa showed increasing resistance to ceftazidime, ciprofloxacin and piperacillin/tazobactam. This was due to an outbreak among rather few patients. The overall resistance rates for Gram-negative bacteria were low for aminoglycosides and imipenem. From 1993 to 1996 the total antibiotic consumption decreased by 27% in the whole hospital and 16.5% in the ICUs. However, the reduced antibiotic consumption was paralleled with a 23% decrease in the total number of patients treated in the hospital from 1993 to 1996. In contrast there was an 11.5% increase in the number of ICU patients treated during this period. The conclusion is that all ICUs within a hospital should have a programme for 'on-line' antibiotic resistance surveillance of drugs used in that unit in order to change the empiric treatment when there is an increase in antibiotic resistance. It is also important to survey the antibiotic consumption in the ICUs in order to avoid further selective pressure on bacteria showing increased resistance rates.     

Influence of chronic Toxoplasma infection on ethylnitrosourea-induced central nervous system tumors in rats

Because prior work with mice had revealed remarkable inhibition of central nervous system (CNS) tumor by chronic Toxoplasma infection, the effect of immunomodulation produced by this obligate intracellular parasite was studied in rats which developed CNS tumors following transplacental exposure to the chemical carcinogen ethylnitrosourea. Groups of Fischer 344 rats which had been exposed to ethylnitrosourea were either uninfected or infected at 1 month of age with a virulent strain of Toxoplasma gondii. Rats were sacrificed when morbid symptoms from tumor growth developed, and neural tissue was prepared for light microscopy. Chronic Toxoplasma infection had no effect on the survival of rats or on the amount, location, or histological type of CNS tumor which developed. Although serum antibody to Toxoplasma was present in all infected rats for the duration of the experiment, there was no histological evidence in the brain of a cellular response to infection or to the presence of tumor. When these results are compared to prior experiments of CNS tumor in mice, they suggest that mechanisms of protection against Toxoplasma infection differ in mice and rats and that an inflammatory component produced by the Toxoplasma organism in the brain is a necessary prerequisite for tumor inhibition.