Metallothionein (MT)-3, originally called growth inhibitory factor (GIF), was initially identified through its ability to inhibit the growth of neuronal cells in the presence of brain extract. MT-3 is the brain specific isoform of the MT family whose specific biological activity associates it with neurological disorders. Indeed, studies report that MT-3 is decreased by ~30% in brains of patients with Alzheimer disease (AD). Furthermore, many lines of evidence suggest that MT-3 engages in specific protein interactions. To address this, we conducted immunoaffinity chromatography experiments using an immobilized anti-mouse MT-3 antibody. We identified five associated proteins from the pool of sixteen recovered using mass spectrometry and tandem mass spectrometry after in-gel trypsin digestion of bands from the affinity chromatography. The proteins identified were: heat shock protein 84 (HSP84), heat shock protein 70 (HSP70), dihydropyrimidinase-like protein-2 (DRP-2), creatine kinase (CK) and beta-actin. Coimmunoprecipitation experiments, also conducted on whole mouse brain extract using the anti-mouse MT-3 antibody along with commercially available antibodies against HSP84 and CK, confirmed that these three proteins were in a single protein complex. Immunohistochemical experiments were then conducted on the perfused mouse brain that confirmed the in situ colocalization of CK and MT-3 in the hippocampus region. These data provide new insights into the involvement of MT-3 in a multiprotein complex, which will be used to understand the biological activity of MT-3 and its role in neurological disease.

Transcobalamin I (TCI) is a vitamin B12 binding protein that is found in the secondary granules of mature neutrophils. The expression of the gene for TCI (TCN1) within neutrophils has been shown to be restricted to the later stages of myeloid development and can therefore be used as a marker for granulocyte differentiation. To study transcriptional control regions important in late stage myeloid gene regulation the genomic sequence for TCN1 has been cloned. Clones were isolated from a genomic library constructed in Charon 4A using homologous full-length cDNA probes. Southern blot analysis showed the gene to reside on five EcoRI fragments totaling 14 kb in length. Two overlapping phage clones, containing the entire 14 kb, were isolated and the introns and exons were mapped using Southern blotting and dideoxy sequencing of subclones. The cDNA is represented by nine exons contained within 12 kb of genomic DNA. Comparison of the genomic structure to gastric intrinsic factor (GIF), another vitamin B12 binding protein, revealed a strikingly similar intron/exon structure, with several positionally conserved splice sites. The gene was localized to chromosome 11 using in situ hybridization.

We previously studied a conditioning paradigm to associate the proboscis extension reflex (PER) with monochromatic light (conditioned stimulus; CS) in harnessed honeybees. Here, we established a novel conditioning paradigm to associate the PER with a motion cue generated using graphics interchange format (GIF) animations with a speed of 12 mm/s speed and a frame rate of 25 Hz as the CS, which were projected onto a screen consisting of a translucent circular cone that largely covered the visual field of the harnessed bee using two liquid crystal projectors. The acquisition rate reached a plateau at approximately 40% after seven trials, indicating that the bees were successfully conditioned with the motion cue. We demonstrated four properties of the conditioning paradigm. First, the acquisition rate was enhanced by antennae deprivation, suggesting that sensory input from the antennae interferes with the visual associative learning. Second, bees conditioned with a backward-direction motion cue did not respond to the forward-direction, suggesting that bees can discriminate the two directions in this paradigm. Third, the bees can retain memory for motion cue direction for 48 h. Finally, the acquisition rate did not differ significantly between foragers and nurse bees.

We investigated if human ocular torsion (OT) and perceived roll (PR) are elicited in response to either dynamic interaural linear acceleration or dynamic roll tilt of the gravito-inertial force (GIF). We expanded a previous study [26] that measured only OT across a limited frequency-range (from 0.35 Hz to 1 Hz) by simultaneously measuring OT and PR at three very low (0.01, 0.02 and 0.05 Hz) and one high (1 Hz) frequencies. Three experimental conditions were investigated: (1) Y-Upright with acceleration along the interaural (Y) axis while upright, (2) Y-Supine with acceleration along the Y-axis while supine, and (3) Z-RED with acceleration along the rostro-caudal Z) axis with right-ear-down (RED). OT was measured by video-oculography, while PR was measured by use of a somatosensory bar. OT and PR were qualitatively different. Large OT responses were measured for Y-Upright and Y-Supine, while large perceived roll responses were observed for Y-Upright and Z-RED. OT for Z-RED was small, and PR for Y-Supine was absent. In conclusion, OT and PR appear governed by qualitatively different neural mechanisms. OT appears mostly influenced by central low-pass filtering of interaural graviceptor cues, while PR appears mostly influenced by roll tilt of the GIF.

BACKGROUND

First-degree family history of early coronary artery disease (CAD) and myocardial infarction (MI) is prognostic among disease-free individuals but may be unreliable. This study evaluated deaths caused by CAD, MI, hypertensive heart disease (HtnHD), and congestive heart failure (CHF) among close and distant relatives.

METHODS

The Utah Population Database contains >2.2 million individual records with genealogy data and 250,000 linked death certificates. Deaths caused by CAD (n = 28,469), MI (n = 26,468), HtnHD (n = 3933), and CHF (n = 11,784) were studied. Familial relative risks (FRRs) were assessed for first- and second-degree relatives. Familiality was also evaluated using the Genealogical Index of Familiality (GIF), which considers close and distant genetic relationships in the Utah Population Database.

RESULTS

Familial relative risks in first-degree (FRR = 1.25, P < .0001) and second-degree (FRR = 1.06, P = .0002) relatives were significant for early age at MI death (<65 years old). Genealogical Index of Familiality analysis demonstrated excess relatedness for deaths caused by MI (case GIF 2.93, mean control GIF 2.73, P < .001) and CHF (2.92 vs 2.66, P < .001). For early age at death, GIFs were significant for MI (3.06 vs 2.54, P < .001), HtnHD (3.22 vs 2.44, P = .003), and CHF (2.64 vs 2.23, P = .003).

CONCLUSIONS

Deaths caused by MI and CHF demonstrate a heritable component in close and distant relatives. For MI, CHF, and HtnHD, for which findings were more pronounced in early age at death, gene discovery may be most effective among early-onset clusters. Excess relatedness was not found for CAD death--perhaps because of heterogeneity within the phenotype--suggesting that this may be a suboptimal phenotype for genetic study.

Host-independent (H-I) derivatives of Bdellovibrio bacteriovorus 109 Davis could not be isolated when concentrated suspensions of host-dependent (H-D) cultures, washed free of spent medium, were plated on host-free media. However, H-I colonies did appear when spent broth was incorporated into the isolation medium, indicating the presence of a factor in the spent medium essential for the growth of H-I cells. This growth factor (GIF) was also present in cell-free extracts of Escherichia coli and a variety of other microorganisms including H-D and H-I derivatives of strain 109 Davis. GIF was heat stable, non-dialyzable, and present in both soluble and particulate fractions of extracts. Heating of extracts at 70 C for 10 min resulted in 10- to 40-fold stimulation in GIF activity, and evidence for a heat-labile inhibitor was obtained. Colonies appearing on host-free medium in these experiments were shown to be those of typical H-I derivatives by isolation and subsequent host-independent cultivation of these organisms. GIF was a conditional requirement dependent on age and size of inoculum for all H-I derivatives characterized. Although GIF stimulated the growth of washed exponential phase cells transferred to fresh medium, it was not essential for growth. However, it was essential for the initiation of growth of washed stationary phase cells from small inocula transferred to fresh medium. It is proposed that GIF is required to initiate growth of metabolically quiescent cells.

OBJECTIVE

Significant portions of the cost and complications of esophagogastroduodenoscopy (EGD) are related to sedation. This study aimed to assess the feasibility, acceptability, and accuracy of unsedated small-caliber transoral EGD (sc-EGD).

METHODS

A 4-phase study was performed in healthy volunteers and patients. Phases 1 and 2 involved assessment of the technical feasibility of sedated sc-EGD and the tolerability of unsedated sc-EGD, respectively, in volunteers. Subsequently, the technical feasibility, tolerability, and diagnostic accuracy of sedated and unsedated sc-EGD were determined by having each patient undergo sc-EGD (Pentax EG-1840) with (phase 3) and without (phase 4) sedation, followed by sedated conventional EGD (c-EGD) (Olympus GIF-100 or GIF-Q140) by a staff endoscopist blinded to the findings of the sc-EGD. The t test for paired samples was used for statistical analysis. A P value of <0.05 was considered significant.

RESULTS

Sedated and unsedated sc-EGD were technically feasible and tolerable in all volunteers. In patients, compared with sedated c-EGD, sedated and unsedated sc-EGD were 96% and 97% accurate, respectively. The overall acceptability of unsedated sc-EGD was only slightly worse than that of sedated c-EGD (median, 2 vs. 1 on a scale of 1-10). After unsedated sc-EGD, 98% of patients expressed willingness to undergo the procedure again. No complications were observed during any phase of the study.

CONCLUSIONS

Unsedated sc-EGD is technically feasible, tolerable, and accurate. It can potentially decrease the costs and complications of sedated conventional EGD.

Glycosylation-inhibiting factor (GIF) is a cytokine that is involved in the regulation of IgE synthesis. The crystal structure of recombinant human GIF was determined by the multiple isomorphous replacement method. The structure was refined to an R factor of 0.168 at 1.9 angstrom resolution. The overall structure is seen to consist of three interconnected subunits forming a barrel with three 6-stranded beta-sheets on the inside and six alpha-helices on the outside. There is a 5-angstrom-diameter "hole" through the middle of the barrel. The barrel structure of GIF in part resembles other "trefoil" cytokines such as interleukin 1 and fibroblast growth factor. Each subunit has a new class of alpha + beta sandwich structure consisting of two beta-alpha-beta motifs. These beta-alpha-beta motifs are related by a pseudo-twofold axis and resemble both interleukin 8 and the peptide binding domain of major histocompatibility complex protein, although the topology of the polypeptide chain is quite different.

The knowledge of brain metallothionein (MT) regulation and especially of MT presence in specific cell types is scarce. Therefore, the effect of several well-known MT inducers, measured by radioimmunoassays using antibodies that cross-react with MT-I and MT-II or specific for MT-I and which do not cross-react with human growth inhibitory factor (GIF or MT-III), has been studied in primary cultures of neurons or astrocytes obtained from rat cerebrum. MT-I levels in glial cells were about ten times higher than those in neuronal cells (538 +/- 194 vs. 49 +/- 16 pg MT-I/micrograms protein, mean +/- S.D. from three separate cell preparations). Increasing the concentration of Zn in the bovine serum albumin (BSA)-containing culture medium up to 50 microM significantly increased MT-I levels by up to 3.5-fold in neurons and 2.5-fold in astrocytes. In contrast, Cu up to 50 microM increased MT-I levels in a saturable manner in both neurons (up to 5-fold) and astrocytes (up to 1.5-fold), the maximum effect occurring at 5 microM Cu. In general, the combination of Zn and Cu further increased MT-I levels. The effect of the metals on MT-I appeared to reflect metal uptake, since MT-I induction was less marked when the BSA concentration in the medium was increased from 2 to 10 mg/ml. Dexamethasone increased MT-I levels in both neurons and astrocytes in vitro in a concentration-dependent manner. Endotoxin, IL-1 and IL-6 did not have a significant effect on glial MT levels at the concentrations studied. The administration of dexamethasone to rats increased MT-I levels in non-frontal cortex, cerebellum, pons+medulla, midbrain and hippocampus, but not in hypothalamus, frontal cortex and striatum. Endotoxin increased liver but not brain MT-I levels. Immunocytochemical studies in adult rat brain preparations with a polyclonal antibody that cross-reacts with MT-I and MT-II indicated that immunostaining was always nuclear in glial cells, whereas in neurons it was nuclear in the cerebral cortex, hippocampus and the granular layer of the cerebellum, and nuclear plus cytoplasmic in Purkinje cells in the cerebellum, hypothalamic nuclei and gigantocellular reticular nucleus in the brain stem. Meninges, choroidal plexus, ependymal and endothelial cells were also MT-immunoreactive.

Neuronal growth inhibitory factor (GIF), a metallothionein-like protein (metallothionein-3), impairs the survival and neurite formation of cultured neurons. Native GIF contains 4 Cu(I) and three Zn(II) ions organized in homometallic metal-thiolate clusters. However, the cluster localization is not known. In this study, the metal-thiolate clusters formed with monovalent and divalent metal ions in the C-terminal domain of human GIF [GIF(32-68)] containing 11 cysteines were investigated. The cluster formation was followed by using electronic absorption, circular dichroism (CD), and magnetic circular dichroism (MCD) spectroscopy, and in the case of Cu(I) complexes also by luminescence spectroscopy at 77 K. Spectroscopic studies on the Cu(I)-GIF(32-68) complexes showed the successive formation of two air-sensitive Cu4S8-9- and Cu6S11-clusters. With Zn(II) and Cd(II) ions, a well-defined M4S11-cluster is formed in which each metal ion is tetrahedrally coordinated by cysteine thiolates. In the 113Cd NMR spectra of 113Cd4-GIF(32-68), recorded at 293 and 323 K, all four 113Cd resonances at 672.8, 620.9, 629.6, and 564.2 ppm were observed only at 323 K. Their detection at elevated temperature indicates a conformational flexibility of this domain. Evidence for the existence of a Cd6-GIF(32-68) complex, contaning two more weakly bound Cd(II) ions, was also obtained. The formation of this complex requires the transformation of some originally terminal thiolates of the Cd4S11-cluster to bridging thiolates, suggesting a more accessible cluster structure. Such properties of Cd4-GIF(32-68) have not been observed with the Cd4S11-cluster in the isolated alpha-domain (amino acids 31-61) of metallothioneins. The significance of Cu- and Zn-clusters for the structure of native GIF is discussed.

Human growth inhibitory factor (GIF) is a new metallothionein-related molecule whose expression is markedly reduced in Alzheimer's disease (AD) brain. We have recently isolated a full-length cDNA for human GIF that has a striking homology to metallothioneins (MTs). As an initial step to better understanding of the biological functions of GIF, we have isolated a full-length cDNA for rat GIF. Comparison of the predicted amino acid sequence with that of human GIF revealed a strikingly high homology (84% identity). Rat GIF cDNA also showed a striking homology to rat MT-1 and MT-II (66% and 62% identities on amino acid sequences, respectively). All cysteine residues were conserved among rat GIF, human GIF and MTs, indicating that cysteine residues play an important function in these molecules. Compared to mammalian MTs, there were 3 bp and 12 bp insertions near the amino-terminal and carboxy-terminal regions in the rat GIF. The rat GIF mRNA was found to be expressed exclusively in the central nervous system, being expressed predominantly in rat cortical astrocytes in primary culture. Although the rat GIF mRNA expression was low during the fetal stage, a dramatic increase of the rat GIF mRNA expression was demonstrated during the postnatal period of 10-17 days. These results indicate that transcriptional regulation of the rat GIF gene is quite different from that of MTs despite its strikingly high homology with MTs.

BACKGROUND

The aim of this study was to assess the yield of antral biopsies performed via unsedated transnasal esophagogastroduodenoscopy, a technique that does not require conscious sedation with its concomitant costs and complications, for documentation of Helicobacter pylori eradication.

METHODS

Nineteen patients who were previously CLO test positive on conventional esophagogastroduodenoscopy and subsequently treated for H pylori infection were enrolled. The subjects had not received antibiotic therapy in the prior month and had no prior gastric surgery. By using a GIF-N30 fiberoptic endoscope and a tiny cup biopsy forceps (1.8 mm diameter), unsedated transnasal endoscopy was performed and antral biopsy specimens were taken for a CLO test, histologic analysis (Dieterle stain), and tissue culture. On the same day, the subjects underwent a carbon 13-labeled area urea breath test. All subjects completed a visual analog scale, rating the acceptability of the unsedated transnasal examination and the previous sedated conventional esophagogastroduodenoscopy.

RESULTS

There was no statistically significant difference between the results of the CLO tests (5/19 positive) versus the 13C-urea breath test (4/19 positive) (p = 0.96), the CLO tests versus histologic findings (5/19 positive) (p = 0.71), or the 13C-urea breath test versus histologic findings (p = 0.96). All tissue culture results were negative. The overall acceptability of unsedated transnasal esophagogastroduodenoscopy was similar to that of sedated conventional esophagogastroduodenoscopy.

CONCLUSIONS

Unsedated transnasal esophagogastroduodenoscopy, a technique that eliminates the costs and complications associated with conscious sedation, is a feasible and accurate alternative to conventional esophagogastroduodenoscopy when documentation of H pylori eradication and confirmation of gastric ulcer healing are both indicated.

Circulating interferon production, induced by Newcastle disease virus, is about seven times higher in C(57) Black mice than i Balb/c/Gif mice. A Mendelian analysis was carried out and circulating interferon production was measured in reciprocal F(1) hybrids, in the F(2) generation, in progeny of backcrosses of F(1) hybrids to either parent strain, and in second backcross progeny. The results indicate that a single, partly dominant, autosomal factor is responsible for the difference in circulating interferon production between both parent strains.

Various drugs are used for sedation prior to upper gastrointestinal endoscopy, some with undesirable side effects. In an attempt to avoid these side effects, 2000 upper diagnostic gastrointestinal endoscopies were performed in a period of 4 years between 1982 and 1986, without any sedation, using Olympus GIF-Q and GIF-P3 gastroscopes. Anxiety, ease of introduction of gastroscope, tolerance of the procedure, and the overall success of the procedure were assessed. Most patients were calm (81.2%); 94.4% had an easy introduction of the gastroscope, 80.3% tolerated the procedure well; and 94.2% of the endoscopies were completely successful. There were no complications, and only four examinations failed (0.2%). Sedation had to be used (intravenous diazepam) in 32 patients due to excessive anxiety and an inability to introduce the gastroscope and in three children under 10 years (1.6%). The average time needed to complete an endoscopy without sedation was found to be 9.5 min, nearly half of the average time needed before this study when sedation was routinely given. It is concluded that upper gastrointestinal endoscopy without sedation can be a safe, quick, well-tolerated procedure.

The phosphorylated signal transduction protein P(II) (P(II)-P) in the cyanobacterium Synechocystis sp. strain PCC 6803 is dephosphorylated by PphA, a protein phosphatase of the 2C family (PP2C). In this study, the physiological conditions of P(II)-P dephosphorylation were investigated with respect to the in vivo specificity of P(II)-P towards PphA and the cellular abundance of PphA in cells growing under different nitrogen regimes. Furthermore, the consequences of impaired P(II)-P dephosphorylation with respect to short-term inhibition of glutamine synthetase (GS) were studied. With a contribution of approximately 15 % of total Mn(2+)-dependent p-nitrophenyl phosphate hydrolysis activity, PphA has only a minor impact on the total PP2C activity in Synechocystis extracts. Nevertheless, residual P(II)-P dephosphorylation in PphA-deficient cells could only be observed after prolonged incubation in the presence of ammonium. The abundance of PphA correlates with the phosphorylation state of P(II) under nitrogen-replete conditions and is specifically enhanced by nitrite. Regulation of pphA expression operates at the post-transcriptional level. In the presence of nitrate/nitrite, PphA is present in molar excess over P(II)-P, enabling the cells to rapidly dephosphorylate P(II)-P in response to changing environmental conditions. A PphA-deficient mutant is not impaired in short-term inhibition of GS activity following ammonium treatment. Down-regulation of GS occurs by induction of gif genes (encoding GS inactivating factors 7 and 17), which is controlled by NtcA-mediated gene repression. Thus, impaired P(II)-P dephosphorylation does not affect ammonium-prompted inactivation of NtcA.

OBJECTIVE

The advent of magnification endoscopy may allow the macroscopic detection of unrecognised villous atrophy in patients with unsuspected coeliac disease. In addition, it may also be possible to use this method to assess the degree of villous atrophy. The aim of this study was to determine the accuracy of zoom endoscopy for the macroscopic evaluation of villous atrophy, in comparison with histological evaluation.

METHODS

The zoom endoscope provided a magnification capability of x 115. A scoring system (Z score) was devised for grading the appearances of villous atrophy: "Z1" for normal mucosa, "Z2" for stunted villi, "Z3" for markedly stunted villi (with ridges and pits) and "Z4" for a flat mucosa. A total of 53 consecutive patients with treated coeliac disease were followed up over almost 2 years using the Olympus GIF-Q240Z zoom endoscope; a total of 80 procedures were carried out. Four biopsies from the second part of the duodenum were taken from each patient for histological assessment. Histological assessment of villous atrophy was made by a pathologist blinded to the Z score. The correlation between the Z score and the histological score was assessed using the weighted kappa method.

RESULTS

The kappa score for the correlation between the macroscopic assessment of villous atrophy and the histology was 0.631, indicating fair to good reproducibility. Agresti's method revealed a very strong baseline association between the two methods ( P < 0.001). Zoom endoscopy had a positive predictive value of 83 % and a negative predictive value of 77 % in detecting villous atrophy.

CONCLUSIONS

Our findings suggest that zoom endoscopy may be valuable in assessing the degree of villous atrophy. However, further studies are needed to assess its efficacy in routine practice as a screening or case-finding tool.

BACKGROUND

Fibroblasts are the principal stromal cells that exist in whole organs and play vital roles in many biological processes. Although the functional diversity of fibroblasts has been estimated, a comprehensive analysis of fibroblasts from the whole body has not been performed and their transcriptional diversity has not been sufficiently explored. The aim of this study was to elucidate the transcriptional diversity of human fibroblasts within the whole body.

METHODS

Global gene expression analysis was performed on 63 human primary fibroblasts from 13 organs. Of these, 32 fibroblasts from gastrointestinal organs (gastrointestinal fibroblasts: GIFs) were obtained from a pair of 2 anatomical sites: the submucosal layer (submucosal fibroblasts: SMFs) and the subperitoneal layer (subperitoneal fibroblasts: SPFs). Using hierarchical clustering analysis, we elucidated identifiable subgroups of fibroblasts and analyzed the transcriptional character of each subgroup.

RESULTS

In unsupervised clustering, 2 major clusters that separate GIFs and non-GIFs were observed. Organ- and anatomical site-dependent clusters within GIFs were also observed. The signature genes that discriminated GIFs from non-GIFs, SMFs from SPFs, and the fibroblasts of one organ from another organ consisted of genes associated with transcriptional regulation, signaling ligands, and extracellular matrix remodeling.

CONCLUSIONS

GIFs are characteristic fibroblasts with specific gene expressions from transcriptional regulation, signaling ligands, and extracellular matrix remodeling related genes. In addition, the anatomical site- and organ-dependent diversity of GIFs was also discovered. These features of GIFs contribute to their specific physiological function and homeostatic maintenance, and create a functional diversity of the gastrointestinal tract.

Multicellular life relies on complex regulatory mechanisms ensuring proper growth and development. In plants, these mechanisms construct a body plan that is both reproducible, and highly flexible for adaptation to different environmental conditions. A crucial regulatory module - consisting of microRNA miR396, GROWTH REGULATING FACTORS (GRFs) and GRF-INTERACTING FACTORS (GIFs) - has been shown to control growth of multiple tissues and organs in a variety of species. Especially in the last few years, research has expanded our knowledge of miR396-GRF/GIF function to crops, where it affects agronomically important traits, and highlighted its role in coordinating growth with endogenous and environmental factors. Special properties make the miR396-GRF/GIF system highly efficient in growth regulation and a promising target for improving plant yield.

Neuronal growth-inhibitory factor (GIF), also named metallothionein-3, inhibits the outgrowth of neuronal cells. Recent studies on the structure of human GIF, carried out using NMR and molecular dynamics simulation techniques, have been summarized. By studying a series of protein-engineered mutants of GIF, we showed that the bioactivity of GIF is modulated by multiple factors, including the unique TCPCP motif-induced characteristic conformation, the solvent accessibility and dynamics of the metal-thiolate cluster, and the domain-domain interactions.

Three orf virus putative virulence proteins are described that exhibit immunomodulatory functions. The OVIFNR gene at the left terminus of the viral genome encodes an interferon resistance protein with homology to the E3L gene of vaccinia virus. OVIFNR functions by preventing a dsRNA-dependent kinase from inhibiting virus and cell protein synthesis as part of the interferon-induced anti-viral state within infected cells. The orf virus orthologue of the ovine interleukin-10 (vIL-10) gene is located at the right terminus of the viral genome. Both vIL-10 and host (ovine) IL-10 function in vitro as inhibitors of pro-inflammatory cytokine production by keratinocytes and macrophages, and both inhibit IFN-gamma production from activated peripheral blood lymphocytes. Both the orf virus vIL-10 and ovine IL-10 stimulate mast cell and thymocyte proliferation. In this respect the orf virus IL-10 differs from Epstein Barr virus IL-10 which does not exhibit cell proliferative activity. Finally, the orf virus GM-CSF inhibitory factor gene (GIF) at the right terminus of the viral genome encodes an inhibitor of GM-CSF that also binds IL-2. Together, these viral proteins are capable of inhibiting key components of the ovine anti-virus immune and inflammatory response.

OBJECTIVE

Endoscopic mucosal resection and submucosal dissection can provide curative endoscopic therapy for Paris type I/II adenomas and node-negative early cancer. No studies have addressed the technical feasibility of retroflexion endoscopic dissection methods for luminal "salvage" therapy in patients considered unresectable using conventional forward-viewing resection.

METHODS

Colonoscopy using an Olympus GIF-XQ240 gastroscope was carried out in 76 patients with Paris type I/II adenomas, early colorectal cancer (CRC), or laterally spreading tumors (LSTs) when the index endoscopist considered the lesion to be unresectable due to retrograde fold involvement. Endoscopic mucosal resection (EMR) and submucosal dissection were carried out using a complete retroflexion technique. Endoscopic and miniprobe 20-MHz or 12.5-MHz ultrasound follow-up data were collected prospectively up to 24 months after the index resection.

RESULTS

Cecal intubation or cannulation to the neoterminal ileum was achieved in 76 (100 %) cases. Forty lesions (53 %) were classified in accordance with the Paris criteria as Is; 16 (21 %) as type II; 10 (13.5 %) as LST-G; and 10 (13.5 %) as LST-NG. Eight lesions (10 %) were excluded from EMR on the basis of endoscopic ultrasound criteria, with 68 of the 76 lesions (89 %) meeting the criteria for endoluminal resection. The median intubation time was 16 min (range 3-32 min). The median resection times were 98 min (range 30 - 242 min), 36 min (range 10-60 min), 172 min (range 20 - 240 min), and 60 min (range 10-116 min) for Paris Is, II, LST-G, and LST-NG lesions, respectively. LST-G morphology was associated with a high median submucosal injection volume in comparison with all other Paris types ( P < 0.05) and with a prolonged resection time ( P < 0.01). Sixty-one patients (94 %) completed the surveillance protocol. Higaki recurrence criteria were met in seven patients (11 %), with six undergoing successful adjunctive endoluminal resection. After 24 months of follow-up, the "cure" rate with endoscopic resection was 60 out of 61 (98 %).

CONCLUSIONS

This is the first prospective study to address the safety and medium-term efficacy of retroflexion endoscopic resection in the colon. When appropriate exclusion criteria are applied, selected patients can receive curative resection using the retroflexion technique. "Salvage" endoluminal therapy may therefore be possible in such cases when surgical resection would otherwise have been required.

The innervation of the cornea of newborn (two day old) and adult rats was investigated using glyoxylic-acid-induced fluorescence (GIF) for catacholamines and subsequent acetylcholinesterase reaction. Fluorescent nerve were observed around the limbal vessels and in the pericorneal nerve plexus, from which they branched towards the central parts of the cornea. The fluorescent corneal nerves were either nonvaricose or had varicosities at intervals of 10 micra. When the animals had been pretreated with nialamide, noradrenaline and propranolol, some fluorescent branching nerve terminals with numerous varicosities also appeared. All fluorescent nerves disappeared two days after ipsilateral superior cervical sympathectomy. When the acetylcholinesterase (AChE) reaction was performed subsequently to the GIF reaction the following nerve types could be identified: 1. nerves containing both catecholamine (CA) fluorescence and AChE, 2. Nerves containing only AChE.

To determine its active site, growth inhibitory factor (GIF), a central nervous system-specific metallothionein-like protein, was digested with trypsin followed by Staphylococcus aureus protease V8 digestion. Of 5 peptide fragments separated from trypsin-digested GIF by reverse-phase high pressure liquid chromatography and gel filtration, only GIFGIFGIFGIFGIFGIFGIFGIFGIFGIF is required for growth inhibitory activity and that folding of the peptide via S-metal bonding is critical for biological activity.