Microvasculature, blood flow, and vasoreactivity in rabbit sinus mucosa after surgery.

OBJECTIVES: To study the circulatory integrity of the sinus mucosa following surgery, the vascular anatomy, blood flow, and vasoconstrictor response of the regenerated microcirculatory network were analyzed. STUDY DESIGN: Forty-six New Zealand White rabbits were operated on unilaterally with either a modified radical operation (MRO) or middle meatal antrostomy (MMA), using the nonoperated sinus for control purposes. After surgery, the animals were left to heal for 1 month. METHODS: Vascular casts were prepared with a low-viscosity methyl methacrylate resin and studied by scanning electron microscopy. Blood flow was measured by means of radiolabeled microspheres (tin 113 [113Sn]). The vasoconstrictor response to oxymetazoline at increasing concentrations was measured with laser Doppler flowmetry. RESULTS: The number of vessels increased significantly in the regenerated mucosa. The vascular casts displayed a rich microcirculation with local signs of angiogenesis. However, there was no difference in blood flow between the operated cavities and their control sides. Following MRO the regenerated mucosa was more sensitive to the vasoconstrictor effect of oxymetazoline, compared with the control side. This difference was not evident in the MMA-operated sinuses. CONCLUSIONS: In this model, surgery does not seriously interfere with the sinus blood flow, although the regenerated mucosa did display an altered vasoreactivity. These findings should be considered in relation to the effects of surgery intended to limit local inflammation and to ensure the ventilation of the sinuses.

Constructing biological knowledge bases by extracting information from text sources.

Recently, there has been much effort in making databases for molecular biology more accessible and interoperable. However, information in text form, such as MEDLINE records, remains a greatly underutilized source of biological information. We have begun a research effort aimed at automatically mapping information from text sources into structured representations, such as knowledge bases. Our approach to this task is to use machine-learning methods to induce routines for extracting facts from text. We describe two learning methods that we have applied to this task--a statistical text classification method, and a relational learning method--and our initial experiments in learning such information-extraction routines. We also present an approach to decreasing the cost of learning information-extraction routines by learning from "weakly" labeled training data.

A radiation hybrid map spanning the entire human X chromosome integrating YACs, genes, and STS markers.

We present a radiation hybrid (RH) map of human Chromosome (Chr) X, using 50 markers on 72 radiation hybrids. The markers, obtained from the consensus map, form a grid spanning the entire chromosome. To check the RH map, the marker order was determined by analysis of presence or absence of retained human DNA fragments in the RHs; the comparison with the consensus showed a similar order. Any STSs, microsatellites, genes, and clones can be positioned and ordered relative to the marker grid. This approach integrates genetic, physical, and large-scale clone mapping and is used to link YAC contigs containing data from various experimental sources.

An integrated YAC map of the human X chromosome.

The human X chromosome is associated with a large number of disease phenotypes, principally because of its unique mode of inheritance that tends to reveal all recessive disorders in males. With the longer term goal of identifying and characterizing most of these genes, we have adopted a chromosome-wide strategy to establish a YAC contig map. We have performed > 3250 inter Alu-PCR product hybridizations to identify overlaps between YAC clones. Positional information associated with many of these YAC clones has been derived from our Reference Library Database and a variety of other public sources. We have constructed a YAC contig map of the X chromosome covering 125 Mb of DNA in 25 contigs and containing 906 YAC clones. These contigs have been verified extensively by FISH and by gel and hybridization fingerprinting techniques. This independently derived map exceeds the coverage of recently reported X chromosome maps built as part of whole-genome YAC maps.

A 13-year report on childhood sinusitis: clinical presentations, predisposing factors and possible means of prevention.

Two hundred and nineteen children with sinusitis treated as in-patients at Huddinge University Hospital during the period 1980-1992 have been reviewed. Epidemiological data, the clinical picture, treatment and complications are described. The prevalence of significant predisposing conditions (such as upper airway allergy, asthma, and immunoglobulin deficiency) has been estimated. Serious sinusitis complications are few, surgery is only rarely required, and previously-recognized important predisposing paediatric conditions are not significantly more common than in the general juvenile population. Improved medication and prevention may have reduced the incidence of serious sinus infections in risk groups today. Children with cystic fibrosis have been reviewed with regard to the necessity of both sinus and nasal polyp surgery. Aggressive medical therapy appears to have reduced their need for sinus surgery as well as polypectomy.

Endoscopic and Caldwell-Luc approaches in chronic maxillary sinusitis: a comparative histopathologic study on preoperative and postoperative mucosal morphology.

The aim of the present investigation was to study the histopathologic mucosal changes occurring in chronic maxillary sinusitis both preoperative and postoperative to functional endoscopic sinus (FES) surgery and the Caldwell-Luc (C-L) operation. Correlations were also sought between the histopathologic parameters and endoscopic findings, as well as patient symptoms. Sixty sinuses with the FES surgery and 55 sinuses with the C-L procedure were studied. The histologic parameters were graded semiquantitatively and compared preoperatively and postoperatively. The C-L operation reduced almost all parameters, whereas after the FES operation only edema and inflammatory cells were significantly reduced. Fibrosis increased postoperatively with both methods. The number of inflammatory cells was closely correlated to a thickened antral mucosa and to purulent secretion. No valid correlations were found when comparing histology with patient symptoms. All in all, histologic considerations suggest that asthmatic patients with severe sinonasal polyposis might benefit from the C-L procedure.

Efficient identification and regional positioning of YAC and cosmid clones to human chromosome 21 by radiation fusion hybrids.

The use of integrated mapping strategies involving bacterial, yeast, and rodent cells as hosts simplifies the construction of maps, which combine long-range order, high resolution, and easy access to the cloned DNA. Radiation-fusion hybrids offer a specially powerful long-range mapping system for human chromosomes. We describe here techniques for establishing a radiation-fusion hybrid map of Chromosome (Chr) 21q and its integration with local information on YAC and cosmid positions.

Physical map of small cell lung cancer deletion region on short arm of human chromosome 3 (3p13-22) based on radiation fusion hybrid analysis.

Deletion of DNA sequences from various regions of the short arm of human chromosome 3 (3p13-14, 3p21, and 3p25) has been observed during the development of a variety of solid tumors, including lung and renal cell carcinomas. In this study we have used a set of radiation fusion hybrids to generate a physical map of chromosome 3p to orient the search for putative tumor suppressor genes. Eighty-six human-hamster radiation fusion hybrids were screened on Southern blots for the retention of 55 human chromosome 3p DNA markers. The high marker density enabled us to identify a set of successively overlapping chromosome fragments in the 3p13-22 area guided by eight markers with previously known order. Twenty-four map intervals were suggested using breakpoints determined by partial fragment overlaps. The final order between the markers derived is consistent with previous information about localizations for 26 of the markers to three larger cytogenetic intervals.

Identification of human chromosome region 3p14.2-21.3-specific YAC clones using Alu-PCR products from a radiation hybrid.

Deletion of DNA sequences from at least three different regions on the short arm of human chromosome 3 (3p13-14, 3p21 and 3p25) are frequently observed during the development of many solid tumors, including lung cancers and renal cell carcinomas. In order to physically characterize the 3p21 region, we previously identified a radiation fusion hybrid that contained about 20 megabases of DNA from chromosome region 3p14.2-p21.3. In this study total Alu-PCR products from this hybrid were used as a probe to isolate 86 yeast artificial chromosomes (YAC) clones from a 620-kb average insert YAC library (ICRF). Sixty-nine Alu-PCR markers, generated from the YACs, and seven PCR primers were used to screen for overlaps between individual clones. Seven contigs were identified encompassing 32 YAC clones. Based on previous information about localization of the PCR primers, the three largest contigs could be assigned to smaller subregions between 3p14.2 and 3p21.3. By this work a large proportion of the 3p14.2-21.3 region is covered with large-insert YAC clones.

Regeneration of maxillary sinus mucosa following surgical removal. Experimental study in rabbits.

A rapid regeneration of the epithelium takes place in the maxillary sinus in rabbits after experimental operative removal of the mucosa. Two weeks postoperatively the previously denuded areas have reepithelialized. The subepithelial glands, however, do not seem to regenerate. The normal sinus mucosa contains numerous serous glands in the lamina propria, but in the regenerated mucosa these glands are replaced by dense connective tissue. Atypical glands and polyp formations are sometimes encountered, but goblet cells are sparse. Furthermore, the sinus cavity on the operated side is reduced in size compared with the nonoperated side because of fibrosis and periosteal reactions including bone degradation and neogenesis. This study indicates that although the mucosa is reepithelialized within 2 weeks, the regeneration of the lamina propria is incomplete, and reactive cellular processes such as bone remodeling, fibroblast proliferation, and formation of polyps and "atypical glands" are characteristic of regenerating mucosa.

Radiation fusion hybrids for human chromosomes 3 and X generated at various irradiation doses.

We have used a gamma-irradiation (2.5-25 krads) cell fusion procedure to generate human-hamster somatic cell hybrids (IHB, irradiated human fragments in B14-150 cells), retaining small fragments derived from human chromosomes 3 and X. By using Alu-element mediated PCR amplification and dot-blot hybridization with human alphoid or total human DNA as probes, 86 positive hybrids were identified and selected for further analysis. Nonisotopic fluorescence in situ hybridization (FISH) with human DNA in a set of eight hybrids demonstrated the presence of from one to eight human fragments per cell independent of irradiation dose. In contrast, a significant dose-dependent variation of fragment sizes was shown in the analysis of the 86 hybrids with markers previously mapped to 3p (seven markers) and to Xq (21 markers). Using the Xq27-28 region as a model, 40% of the hybrids generated at 5 krads or less were found to have retained fragments in the range of 3-30 Mb, 10% retained the whole chromosome arm, and the remaining 50% retained fragments of less than 2-3 Mb. The proportion of fragments of 3 Mb or larger decreased rapidly at higher irradiation doses and was very low (less than 6%) in hybrids generated at 25 krads. Upon further characterization, the 86 hybrids analyzed here will provide a mapping panel for the entire chromosomes 3 and X with an estimated resolution in the range of 1-2 Mb on average, a size range amenable to PFGE and YAC contig mapping.

Cellular regeneration and recovery of the maxillary sinus mucosa. An experimental study in rabbits.

Unilateral maxillary sinusitis was induced in 30 New Zealand White rabbits with Streptococcus pneumoniae or Bacteroides fragilis. In another group of 15 rabbits without infection, the sinus mucosa was surgically removed in defined areas. In both series, the sinuses were serially sectioned for histological analysis of the cellular regenerative capacity. In maxillary sinusitis induced by Bacteroides fragilis, an inflammatory and also reparative process involving all mucosal layers including the underlying periosteum was seen. The more superficial trauma as found in pneumococcal sinusitis eventually led to restitution ad integrum. Following surgical removal, the denuded sinus-lining was reepithelized by a flattened ciliated epithelium on a lamina propria displaying fibrosis and lacking serous glands. The restoration of the rabbit maxillary sinus mucosa after surgical trauma thus leads to structural abnormalities of the epithelium as well as the lamina propria, and these changes are likely to interfere with the normal function of the sinus mucosa.

Experimental sinusitis in rabbits induced by aerobic and anaerobic bacteria: models for research in sinusitis.

Experimental acute sinusitis can be induced in New Zealand white rabbits by Streptococcus pneumoniae serotype 3, or the anaerobe Bacteroides fragilis, after blocking of the sinus ostium. Histological examination of the sinus mucosa in both models reveals edema, dilated venules, leukocytic infiltration, goblet cell formation, as well as localized epithelial lesions. In comparison the bacteroides sinusitis enhances a more pronounced and long-lasting tissue reaction, including periosteal thickening and new bone formation. The sinus mucosal blood flow as measured with microspheres Sn113 is increased as compared to control side in pneumococcal sinusitis. An increased lactate concentration of sinus secretions as well as in the pathological sinus mucosa indicates an anaerobic metabolism. Furthermore, a decreased ATP-content of the sinus mucosa suggests an energy depletion which could impair epithelial function. The anaerobic milieu of the sinus secretion is probably created by the leukocytes as analyzed by the separation of the D- and L-form of lactate.

Autoradiographic findings in experimental sinusitis in rabbits.

In order to study the effect of a purulent infection in the sinus cavity on the uptake of radionuclide in the surrounding bone, two series of experiments were performed; one using 45Ca-chloride (T1/2 = 153 d), that was given intravenously in a dose of 50 MBq to the previously infected animal, and in the other 99Tcm-DP was used in a dose of 530-1600 MBq. The animals were killed 4-6 hours later and analyzed using an autoradiographic technique and histological examination. Chronic unilateral infection of the maxillary sinus was induced experimentally according to the technique described by Kumlien and Schiratzki (1985). An acute unilateral maxillary sinusitis was induced experimentally according to Johansson et al. (1988). An increased uptake of 99Tcm-DP in the bone surrounding the sinus cavity with an infected mucosa could be seen. This uptake could be seen four days after induction of an acute, purulent, pneumococcal sinusitis, and five months after the induction of an purulent, chronic sinusitis. No increase in uptake could be seen in the mucosa of the sinus with purulent infection. The beta-emitting nuclide gave better resolution in autoradiography than did the gamma-emitting nuclide, and would give better possibilities in identifying structures; however, the specificity to bone of 45Ca was not high enough to be suitable.

Analysis of a glucose-containing tetrasaccharide by high-performance liquid affinity chromatography.

In the present work we have explored conditions for using a pulsed amperometric detector for on-line analysis of oligosaccharides eluted from a high-performance liquid affinity chromatography column. A monoclonal antibody that specifically binds a glucose-containing oligosaccharide is coupled to a SelectiSphere-10-activated tresyl column. The system is eluted isocratically and easily detects 10 ng of the oligosaccharide with a linear response up to 250 ng. Analysis of both serum and urine samples from normal individuals and patients with acute pancreatitis gives a single retarded peak with a retention time identical to that of authentic (Glc)4. Retarded material pooled from several analyses of urine was positively identified as (Glc)4 by GC-MS analysis. As this method requires little cleanup and no chemical derivitization of the sample and is performed rapidly (less than 20 min) at sensitivities of at least 10 micrograms/liter in biological fluids, it represents a substantial improvement over previous GC-MS, radioimmunoassay, and enzyme-linked immunoadsorbent assay methods used to determine (Glc)4.

Determination of a glucose-containing tetrasaccharide in urine of patients with acute pancreatitis.

The oligosaccharide Glc alpha 1-6Glc alpha 1-4Glc alpha 1-4Glc (Glc4), formed by amylolytic degradation of starch and glycogen, is excreted at extremely high levels in the urine of patients with acute pancreatitis. To determine if Glc4 is a useful indicator for this disease, we studied 55 patients admitted to the hospital because of acute abdominal pain (possibly acute pancreatitis). The highest values of Glc4 excretion in urine was found in a group of 11 patients with acute pancreatitis. The Glc4 values, but not serum and urine amylase values obtained at admission, correlated significantly with the severity of the disease. In two patients, followed through convalescence, the Glc4 excretion remained elevated for 7 and 2 wk in contrast to serum and urine amylase that returned to normal levels within a few days. It seems probable that glycogen, released from the liver and subsequently degraded by amylase originating from the pancreas, can be measured as Glc4 in the urine. This might be valuable for diagnosing acute pancreatitis and gives indications of the prognosis.