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PubMed2001

Proximal bronchial extension with special reference to tumor localization in non-small cell lung cancer.

OBJECTIVE: Surgery is the optimal treatment in patients with non-small cell lung cancer (NSCLC) and tumor-negative bronchial resection margins should be maintained for a curative resection. The epidemiology of NSCLC, including the aspects of tumor localization, has been changing during the recent decades. The aim of this study was to evaluate microscopic proximal bronchial extension with special reference to the site of the tumor. METHODS: Surgical specimens of 70 NSCLC cases were examined histologically for proximal bronchial extension of the tumor. The entire bronchial tree with the tumor was extracted from the specimen and serially cut at a thickness of 5 mm in the transverse plane of the bronchus. Microscopic proximal extension of the tumor was classified as either endobronchial or peribronchial. RESULTS: Thirty-three (47.1%) tumors had central and 37 (52.9%) had peripheral localization. Among the central and peripheral tumors, 10 (30.3%) and seven (18.9%) had microscopic proximal extension, respectively. In total, the mean length of proximal extension was 10.94 +/- 7.07 mm. The mean length of extension for peripheral tumors was 15.71 +/- 8.38 mm, significantly greater than that of central tumors, which was 7.60 +/- 3.47 mm (P = 0.026). Peripheral tumors showed a significant peribronchial extension (P = 0.024). CONCLUSIONS: A greater percentage of central tumors show microscopic proximal bronchial extension, whereas the length of microscopic proximal bronchial extension is significantly greater in peripheral tumors. Peripheral tumors preferentially have a peribronchial extension pattern.

Adenocarcinoma↗
PubMedDate not supplied

Computer-assisted, operator-interactive technique for calculating pulmonary arterial taper.

We have developed a computer-assisted, operator-interactive technique which performs fast, precise computations of pulmonary artery taper. Individual 35-mm cineframes from balloon-occlusion pulmonary arteriograms are digitized into a 640 X 480 matrix in 8-bit depth and loaded into a VAX 11/780 computer for analysis. After operator identification of the arterial segment, an automated process of caliber analysis is initiated. By fitting a cubic spline function to the densitometric profiles extracted from the arterial segment, serial arterial cross-sectional diameters are calculated from the mathematically-derived points along the fitted curves. Spurious profiles, caused by sectioning at bifurcations, can be overridden by an operator-interactive subroutine. Taper is derived from the slope of the least-squares fit of vessel caliber with respect to its distance along the arterial segment. Results obtained by calculations from the computer-assisted caliber measurements were compared with those obtained by hand-tracing the same vessel segments. Correlation between computer-traced inflection points and hand-traced taper was very significant (r = .96, n = 13, P less than 0.001).

Child, Preschool↗
PubMed1991

Effect of endogenous digitalis-like factor on endothelin secretion from bovine endothelial cells.

We have recently isolated two endogenous digitalis-like factors (EDLFs) from human urine. The polar EDLF (ouabain-displacing compound 1; ODC-1) fulfills the criteria for the putative natriuretic and vasoactive digitalis-like factor. In this study, we examined the effects of ODC-1 and ouabain on endothelin-1 (ET-1) secretion from cultured endothelial cells. ODC-1 was partially purified from human urine with use of reverse-phase HPLC based on the inhibitory effects on [3H]ouabain binding to intact human erythrocytes. A serial dilution curve of the extract of the serum-free culture medium from CPAE (bovine pulmonary artery endothelium) cells was parallel to that of standard ET-1. Immunoreactive (ir-) ET-1 was secreted in a linear fashion over 5 h from CPAE cells. ODC-1 stimulated ET-1 secretion in a dose-dependent manner. In contrast, ouabain did not affect ET-1 secretion at the concentration of 10(-9)-10(-5) M. These results indicate that human urine-derived ODC-1 is distinct from plant-derived ouabain and acts as a stimulator of ET-1 secretion, and also suggest that ODC-1 may play an important role as a direct and indirect modulator of vascular tone.

Animals↗
PubMed1968

Labeling pattern of translocated stachyose in squash.

One mature blade of each squash plant was continuously labeled with (14)CO(2) for 15, 30, or 70 minutes in light. The ethanol soluble materials from serial sections of petioles were extracted and separated by paper chromatography. The ratios of label in the various components of this fraction were determined. Stachyose, which contained the major portion of the label of this fraction, was hydrolyzed and the resultant hydrolysate was separated by paper chromatography. Specific activities of the hexoses derived from stachyose were determined. It was found that the glucose and fructose moieties of stachyose became labeled at the same rates; however, the galactose moiety became labeled more rapidly.

Journal Article↗
PubMed1996

Detection of gastric cancer micrometastases in lymph nodes by amplification of keratin 19 mRNA with reverse transcriptase-polymerase chain reaction.

A sensitive method for the detection of gastric cancer micrometastases in lymph nodes was developed. The method was based on amplification of keratin 19 mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR). Keratin 19 RT-PCR showed that keratin 19 mRNA was expressed in all 12 gastric cancers, but not in any of 20 normal control lymph nodes, indicating that keratin 19 mRNA is a good target of RT-PCR for the detection of gastric cancer micrometastases in lymph nodes. Serial dilution studies of RNA extracted from gastric cancers against RNA extracted from control lymph nodes demonstrated that the detection sensitivity of the keratin 19 RT-PCR method was one cancer cell in 10(3)-10(5) lymph node cells. Detectability of lymph node metastases was compared between keratin 19 RT-PCR and conventional histological examination, using 100 lymph nodes obtained from 12 gastric cancer patients. Keratin 19 mRNA was detected in all of the seven lymph nodes which were histologically metastasis-positive. Of the 93 lymph nodes which were histologically metastasis-negative, 79 were found not to express keratin 19 mRNA but 14 were found to express keratin 19 mRNA, indicating that these lymph nodes contained micrometastases which could not be detected by histological examination. These results demonstrate that keratin 19 RT-PCR is a more sensitive method than histological examination for the detection of gastric micrometastases in lymph nodes.

Adult↗
PubMed1995

Clinical diagnosis of internal resorption: an exception to the rule.

This paper describes a case in which the radiographic appearance of a resorptive lesion in a mandibular incisor tooth posed difficulty in the diagnosis of its origin. The lesion did not conform to the normally accepted criteria for diagnosis of internal resorption. Clinical examination did not support the diagnosis of external cervical resorption. Serial histological section of the extracted tooth subsequently confirmed the diagnosis of internal resorption. The criteria for differential diagnosis of resorptive lesions with smooth outlines in the cervical regions of teeth are discussed together with the rationale for their management.

Adult↗
PubMed1999

Adaptation and population dynamics of Azotobacter vinelandii during aerobic biological treatment of olive-mill wastewater.

Olive-mill wastewater (OMW) has a high organic and polyphenol content and is resistant to biodegradation. Its disposal leads to a major environmental pollution problem in the Mediterranean basin. The detoxification of OMW following inoculation with Azotobacter vinelandii (strain A) was performed for two successive 5-day-period cycles in an aerobic, biowheel-type reactor, under non-sterile conditions. The phytotoxicity of the processed product was reduced by over 90% at the end of both cycles. To exclusively monitor the A. vinelandii population in the reactor a most probable number-PCR approach was employed and applied daily to serial dilutions of total DNA extracted from reactor samples. PCR sensitivity was independent of the presence of OMW or non-target DNA. The A. vinelandii population dynamics were successfully monitored, showing an initial adaptation period, followed by a sharp population maximum on the fourth day of both cycles (1.6x10(8) and 9.6x10(7) cells ml(-1) respectively), after a major phytotoxicity decline. N(2) fixation rates were estimated using the acetylene reduction assay and reached a peak during the first 1-2 days of each cycle (36 and 29 nmol C(2)H(2) ml(-1) h(-1) respectively). The data are consistent with an initial physiological adaptation phase, where the presence of phenolic compounds limits A. vinelandii growth but stimulates N(2) fixation, followed by a rapid growth phase as phytotoxicity declines.

Journal Article↗
PubMed1992

Morphology of neural endings in the human periodontal ligament: an electron microscopic study.

The ultrastructure of sensory nerve endings in the human periodontal ligament from 43 extracted teeth was studied using serial sections. Three types of nerve endings were found: free nerve endings (FNE), Ruffini-like endings and lamellated corpuscles. Free nerve endings stem from unmyelinated or from myelinated nerve fibers. The endings contain neurotubuli, neurofilaments and vesicles. Ruffini-like receptors were mostly found in the apical part of the periodontal ligament. In these Ruffini-like receptors a particularly abundant concentration of mitochondria appears. In some cases desmosome-like junctions are present between neurite and ensheathing cell. Lamellated corpuscles were also found in the periodontal ligament. The lamellae are extremely endocytotic and are in close contact with each other.

Adolescent↗
PubMed1994

Electrophoretic transfer from polyacrylamide gel to nitrocellulose sheets, a new method to characterize multilocus enzyme genotypes of Klebsiella strains.

A new method for multilocus enzyme electrophoresis, based on electrophoretic transfers to nitrocellulose after polyacrylamide-agarose gel electrophoresis was explored. Electrophoretic separation was performed on 1-mm-thick slab gels with 6-mul samples of bacterial extracts and was followed by serial 5-min consecutive transfers. The transferability of 19 metabolic enzymes of Klebsiella strains was studied and allowed the simultaneous examination of one enzyme in the separation gel and at least five enzymes on nitrocellulose sheets. The resolution of enzyme bands was increased on nitrocellulose; thus, well-separated bands were recorded for nucleoside phosphorylase, peptidase, and phosphoglucose isomerase whereas their mobility variants could not be clearly distinguished in the separation gel because of stain diffusion. The study of genetic relationships of 42 strains of Klebsiella pneumoniae and 24 strains of Klebsiella oxytoca demonstrated the reliability of the method, since clustering analysis of electrophoretic types, based on electrophoretic polymorphism of 10 metabolic enzymes, showed two main clusters well correlated with the two species. The 57 electrophoretic types described confirm the usefulness of the method for the study of genetic relationships between closely related strains.

Journal Article↗
PubMed2026

Long-acting reversible contraception, condom use and sexually transmitted infections in developing countries: a multi-country serial cross-sectional analysis using demographic and health surveys.

INTRODUCTION: Unintended pregnancy and sexually transmitted infections (STIs) are major public health issues in developing countries. While long-acting reversible contraception (LARC) effectively prevents unintended pregnancy, there is limited evidence from large multinational studies on its association with condom use and STI-related outcomes. This study aimed to investigate the association between LARC use, condom use and STIs among women in developing countries. METHODS: This serial cross-sectional study extracted data from Demographic and Health Surveys (DHS), a series of nationally representative household surveys conducted in developing countries. The analysis included women aged 15 to 49 years, with data on contraceptive methods and demographics. Generalised linear mixed effect models (GLMMs) were used to estimate adjusted prevalence ratios (aPRs) for condom use and self-reported STI-related outcomes, comparing LARC users with both non-LARC users and oral contraceptive users. Subgroup analyses were conducted at the individual level and at the country level. RESULTS: Data from 2 171 884 women across 31 countries were analysed. Overall, the prevalence of self-reported STI-related outcomes was 7.4%, 3.9% of participants used LARC, and 7.5% of participants reported consistent condom use. LARC users were significantly less likely to use condoms compared with non-LARC users (aPR=0.40, 95% CI 0.30 to 0.53) and compared with oral contraceptive users (aPR=0.61, 95% CI 0.48 to 0.78). LARC use was associated with a higher prevalence of STI-related outcomes compared with non-LARC users (aPR=1.19, 95% CI 1.10 to 1.28) and compared with oral contraceptives users (aPR=1.14, 95% CI 1.05 to 1.24). Associations were stronger in low-Human Development Index (HDI) countries, especially among younger women (15-19 years), but were not significant in high-HDI countries. Country-level heterogeneity was observed. CONCLUSIONS: LARC use is associated with reduced condom use and higher self-reported STI prevalence, particularly among younger women and in lower HDI developing countries. These findings support integrating STI prevention into LARC services and promoting dual-method use to prevent both unintended pregnancies and STIs.

Humans↗
PubMed1994

Fulminant hepatic failure resulting from coexistent Wilson's disease and hepatitis E.

Fulminant hepatic failure resulting from hepatitis E and coexistent Wilson's disease was diagnosed in a six year old girl six weeks after returning from a holiday in India. Wilson's disease was diagnosed on the basis of histological evidence of hepatocellular copper deposition, confirmed by biochemical estimation of liver copper concentration. Although severely damaged, the liver was non-cirrhotic. Hepatitis E virus (HEV) was diagnosed by nested polymerase chain reaction, the specificity of which was confirmed by direct sequencing of amplified DNA. Replication of HEV within the liver at the time of diagnosis was confirmed by selective amplification of the antigenomic strand of the virus obtained from total liver RNA. The patient had an orthotopic liver transplantation without recurrence of hepatitis and remains well at 19 months. Viral excretion, recorded by serial amplification of HEV RNA extracted from stool samples, persisted for 30 days after liver grafting. Severe vitiligo, present preoperatively, dramatically improved after liver grafting and institution of immunosuppressive treatment. This case suggests that viral infection may play a part in the acute decompensation seen in some cases of Wilson's disease.

Base Sequence↗
PubMed1976

Synthesis of LH-RH by rat hypothalamic tissue in vitro: I. Use of a specific antibody to LH-RH for immunoprecipitation.

Following the procedure of Jeffcoate et al. [1974] we have successfully obtained a specific antiserum to LH-RH in rabbits, and are utilizing our antibody to study the synthesis of LH-RH by rat hypothalamic tissues in vitro. Our antibody, used at a dilution of 1:20,000, binds 37% of added 125I-labelled LH-RH, and cross-reacts minimally with thyrotropin releasing factor (TRF; 4.0%), somatotropin release inhibiting factor (SRIF; 0.035%), and melanocyte inhibiting factor (MIF; 0.009%). The antiserum recovers LH-RH added to charcoal-stripped rat serum quantitatively. Serial dilutions of rat hypothalamic extract and varying aliquots of normal rat serum are parallel to the standard curve (synthetic LH-RH: 0-100 pg/tube). Hypothalamic tissue of 4 male rats, bounded by the optic chiasm, mammillary bodies and hypothalamic fissures to a depth of 2-3 mm, was pooled, minced and incubated for 1-3 h in 2 ml of Eagle's Minimum Essential Medium in atmosphere of 95% O2/5% CO2 with 10 muCi of 3H-glycine. Reactions were stopped by the addition of 1 ml 0.1 N HC1. The tissue and medium were homogenized together and boiled for 3 min. Aliquots from the incubates were neutralized, LH-RH levels measured by radioimmunoassay (RIA), and 3H-glycine incorporation determined by immunoprecipitation, using our antibody. 3H-glycine incorporation into presumptive LH-RH increased linearly over the 3 h period. Aliquots of the 3 h incubates were chromatographed on Sephadex G-25 columns (1 x 10 cm) using 0.01 M acetic acid for elution; 1 ml fractions were collected. Synthetic LH-RH was chromatographed in a similar manner, and its elution profile determined by UV absorbance at 280 mm. Aliquots of each fraction of the eluted material from the 3 h incubates were counted in a scintillation counter to determine the elution pattern of the labelled material. Additionally, aliquots from the 1 ml fractions were neutralized and used for immunoprecipitation. Coincident peaks of 3H-label, immunoprecipitable 3H-labelled presumptive LH-RH, and synthetic LH-RH were observed. These results lend strong evidence to support our conclusion that the technique of immunoprecipitation is an efficacious approach to the study of the synthesis of LH-RH by rat hypothalamic tissue in vitro.

Animals↗
PubMed1994

Impact of the validator and the validation method on the outcome of occlusal caries diagnosis.

In studies evaluating the performance of caries-diagnostic methods, a validation of the true state of disease is needed. The aim of this study was to evaluate the impact of the validator and the validation system (stereomicroscopy or radiography of tooth sections) on the outcome of diagnostic tests for occlusal caries. The material consisted of 60 extracted third molars which were serially sectioned (500-600 microns thick). Four observers examined the sections by two caries validation methods: stereomicroscopy and film radiography. The presence of caries in the occlusal surfaces of these teeth had previously been recorded by visual inspection and conventional film radiography. The kappa values for interobserver agreement within one validation method ranged from 0.44 to 0.76 for radiography and from 0.47 to 0.60 for microscopy. The intraobserver agreement with the two methods was low (range 0.31-0.49), and by cross-tabulating the data, it was found that the disagreements originated in a consistently deeper lesion score with stereomicroscopy than with radiography by all observers except 1. By use of receiver operating characteristic curve areas, little impact of the validation method was seen when visual inspection was validated (against microscopy mean area = 0.75, against radiography mean area = 0.74). The mean receiver operating characteristic area was higher when diagnostic film radiographs were validated against radiography (0.68) than against microscopy (0.63). The differences between observers within a validation method were larger with microscopy than with radiography. In conclusion, caries validation methods are subject to variability. The outcome of caries--diagnostic tests may be influenced both by the validator and the validation method.

Dental Caries↗
PubMed1984

Oxytocin in the corpus luteum of the cynomolgus monkey (Macaca fascicularis).

To determine if oxytocin (OT) is present in cynomolgus monkey corpus luteum, OT was measured by a specific and sensitive RIA in 13 corpora lutea, ovarian venous plasma on the ipsilateral side and peripheral venous plasma at different stages of the luteal phase. Serial dilution of acetic acid extract of the corpus luteum showed parallelism with standard OT in the RIA. Total content of OT in corpus luteum was 1.9 +/- 0.5 ng (mean +/- SEM) with a content of 0.4-0.8 ng in early luteal phase, 1.0-6.2 ng in midluteal phase, and 0.4-0.7 ng in late luteal phase. OT concentrations in corpus luteum were 21.0-75.2 ng/g wet wt in early luteal phase, increasing to 34.4-602.5 ng/g in midluteal phase; and declining to 3.4-117.4 ng/g in late luteal phase. OT concentrations per mg protein in the corpus luteum were 0.05-19.6 ng with peak concentrations of 14.7-19.6 ng/mg protein on day 22. Sephadex G-25 column chromatography of the corpus luteum extract revealed a single peak for binding activity similar to that of synthetic OT on the RIA. Ovarian vein blood from the same side as the corpus luteum had a significantly higher OT concentrations of 161.2 +/- 29.7 pg/ml on days 15-24 than 16.8 +/- 3.6 pg/ml on days 25-28 (P less than 0.01) and peripheral plasma OT levels of 23.2 +/- 3.4 pg/ml (P less than 0.025). Our findings indicate that OT is present and probably produced by monkey corpus luteum with peak OT concentrations found in midluteal phase. Thus OT may play a role in primate corpus luteum function.

Animals↗
PubMed1997

Characterization of the atrial natriuretic peptide system in the oviduct.

The atrial natriuretic peptide (ANP) gene is expressed in several extraatrial tissues in which ANP may be involved in the regulation of autocrine or paracrine functions. In the ovary, the synthesis of ANP, its binding sites, and a physiological role were found. The ANP system in the oviduct, however, is yet to be defined. The purpose of the present study was to determine whether the ANP system is present in the oviduct and then to define its function. The serial dilution curves of oviductal extracts in rat and rabbit were parallel to the standard curve of ANP. Molecular profiles using reverse phase HPLC indicated that the prohormone and processed circulating peptide were the main forms present. The immunoreactive ANP content of the oviduct was 27.07 +/- 4.41 pg/mg tissue wet wt (1.19 +/- 0.19 ng/oviduct; n = 10; at metestrus) in rats and 1.21 +/- 0.12 pg/mg tissue wet wt (0.15 +/- 0.01 ng/oviduct; n 12) in rabbits. In adult 4-day cycling rats, the immunoreactive ANP contents in oviducts had a cyclic change characterized by the lowest level at proestrus (14.59 +/- 3.24 pg/mg; n = 12). A distinct and strong ANP immunoreactivity was found in the mucosal layer of rat oviduct, and ANP messenger RNA was also detected in the oviduct by reverse transcriptase-PCR. Specific high affinity binding sites for iodinated rat ANP ([125I]rANP) were observed in the mucosal layer of the oviduct in rats and rabbits. Specific [125I]rANP bindings localized in the mucosal layer of rabbit oviduct showed an apparent dissociation constant (Kd) of 18.69 +/- 5.55 nM and a maximal binding capacity of 14.85 +/- 6.19 fmol/mm2. These specific [125I]rANP bindings were not reversed by des-[Gln18,Ser19,Gly20,Leu21,Gly22]ANP-(4- 23) as a selective ligand of clearance receptor. Synthetic ANP inhibited both the frequency and amplitude of basal motility of rabbit oviduct in a dose-dependent manner. These results suggest that the oviduct has its own ANP system, and the system is involved in the regulation of oviductal motility.

Animals↗
PubMed1978

Enzyme-linked immunoassay of somatostatin.

An enzyme-linked immunoassay for somatostatin using somatostatin-alkaline phosphatase conjugate as "labeled" antigen was developed. Minimal detectable dose at present was 40 pg per tube. Serial dilutions of rat hypothalamic extract gave a gradual change of antibody-bound alkaline phosphatase activity which was parallel to that with standard somatostatin. Precision and accuracy of the method were comparable to those in radioimmunoassay reported by others. This method will be a useful tool for the determination of somatostatin, especially in tissues.

Alkaline Phosphatase↗
PubMed1986

Determination of insulin-like growth factor-I in normal subjects and in patients with growth hormone disorders by radioimmunoassay using biosynthetic homologous peptide.

A highly sensitive and specific RIA for IGF-I has been developed using recombinant DNA-derived IGF-I of very high purity and specific antiserum to it. This assay system could detect IGF-I at as low concentrations as 20-30 ng/ml. The intra-assay and interassay coefficients of variation at various concentrations of IGF-I were 4.9 to 6.5% and 5.4 to 8.0%, respectively. The recovery rate of pure IGF-I added to plasma was 77.0 +/- 3.7%. The antiserum did not cross-react with porcine insulin, biosynthetic human insulin, hGH, hEGF, the synthetic C-domain of IGF-I or that of IGF-II, but reacted equally with an analog, Thr59-IGF-I. Plasma IGF-I was extracted by the acid-ethanol method before assay to separate IGF-I from its binding protein. When plasma IGF-I was assayed without extraction, the inhibition curves of serial dilution of plasma samples from several individuals were not parallel to the standard curve of IGF-I. The plasma concentration of IGF-I was 147 +/- 49 ng/ml (mean +/- SD) in 156 normal adults aged from 20-59 years. As reported by others, the IGF-I levels were low in cord plasma (41.8 +/- 23.5 ng/ml) and plasma of patients with GH deficiency (64.6 +/- 42.0 ng/ml), while its levels were high in normal children of pubertal ages (12-13 yr, 365 +/- 126 ng/ml) and in patients with active acromegaly (562 +/- 115 ng/ml). This RIA system is a simple and useful method for determining plasma IGF-I in normal and diseased states.

Acromegaly↗
PubMed2000

Coexistence of C-type natriuretic peptide and atrial natriuretic peptide systems in the bovine cornea.

PURPOSE: To determine whether the cornea synthesizes natriuretic peptides and contains their receptors. METHODS: The synthesis of the natriuretic peptides, C-type natriuretic peptide (CNP) and atrial natriuretic peptide (ANP), in the bovine cornea was determined by high-performance liquid chromatography (HPLC) with radioimmunoassay and Southern blot analysis. The presence of natriuretic peptide receptor (NPR)-A and -B and their localizations were measured by reverse transcription-polymerase chain reaction (RT-PCR), in vitro autoradiography, and the activation of particulate guanylyl cyclase by natriuretic peptides in the corneal membrane. RESULTS: The serial dilution curves of corneal extracts were parallel to the standard curves of CNP and ANP. With reversed-phase HPLC, a major immunoreactive peak of CNP or ANP was observed at the elution time corresponding with synthetic CNP(1-53) or atriopeptin III (APIII), respectively. The presence of mRNAs of CNP and ANP was also detected in the cornea by RT-PCR and/or Southern blot analysis. Production of 3',5'-cyclic guanosine monophosphate (cGMP) by the activation of particulate guanylyl cyclase in the corneal membrane was stimulated by ANP, BNP, and CNP. More cGMP was produced by CNP than by the other natriuretic peptides. Specific 125I-[Tyr0]-CNP(1-22) binding sites were localized in the endothelial cell layer of cornea. The apparent dissociation constant (Kd) value of the cornea was 3.06 +/- 0.73 nM and the maximum binding capacity was 3.40 +/- 0.63 femtomoles/mm2. Both NPR-A and NPR-B mRNAs were detected by RT-PCR. CONCLUSIONS: The cornea synthesizes CNP and ANP and contains their receptors. These results suggest that the CNP and ANP systems coexist in the bovine cornea.

Animals↗