Presence of active gelatinases in endometrial carcinoma and correlation of matrix metalloproteinase expression with increasing tumor grade and invasion

BACKGROUND: The actions of the extracellular-matrix degrading enzymes, matrix metalloproteinases (MMPs), are implicated in tumorigenesis. The cellular localization of MMP-2, MMP-9, membrane type 1 (MT1)-MMP, tissue inhibitors of metalloproteinases (TIMPs) 1-3, and the presence of active gelatinases were investigated in endometrial carcinoma. METHODS: Endometrial carcinomas were grouped according to histologic grade (Grades 1-3), depth of myometrial invasion (0, < 50%, > 50%) and the presence of vascular/lymphatic invasion. Twenty-nine endometrial carcinoma biopsies were investigated immunohistochemically to determine the tissue localization of MMP-2 (gelatinase A), MMP-9 (gelatinase B), MT1-MMP, and TIMPs 1-3. In situ hybridization was performed to localize MMP-2 and MMP-9 mRNA. The presence of active gelatinases was assessed using in situ zymography. RESULTS: Epithelial tumor cells were the main site of MMP-2, MMP-9, and MT1-MMP protein. Variable stromal cell localization was also observed, particularly in areas adjacent to tumor nests. Semiquantitative analysis revealed increases in MMP-9 and MMP-2 but not MT1-MMP staining scores in tumor epithelial cells in the transition from histologic Grade 1 to Grades 2 and 3. Matrix metalloproteinase-9 and MT1-MMP staining scores in tumor cells were significantly associated with the presence of myometrial invasion and vascular/lymphatic invasion, while MMP-2 did not correlate with these factors. In addition, MT1-MMP was co-localized with MMP-2, supporting its role in the activation of proMMP-2. Tumor cells from all histologic grades stained intensely for TIMP-2 and TIMP-3 proteins, while variable stromal staining was observed. In Grade 1 carcinomas TIMP-1 was predominantly immunolocalized to the stromal compartment with variable tumor cell localization being observed in Grades 2 and 3 carcinomas. Matrix metalloproteinase-9 and MMP-2 mRNAs were predominantly observed in tumor epithelial cells as well as in the stroma to varying degrees. In situ zymography revealed active forms of gelatinases at the cellular surface and in association with tumor epithelial cells within endometrial carcinoma tissues. CONCLUSIONS: These data suggest that increasing expression of MMPs and endometrial carcinoma progression are closely related. Active gelatinases are present in endometrial carcinoma, resulting in alterations to the microenvironment that promote tumor invasion and metastasis.     

Accelerated pubertal development in patients with shunted hydrocephalus

OBJECTIVE: To evaluate pubertal development and peripheral concentrations of gonadotrophins and sex hormones in children with shunted hydrocephalus compared with healthy controls. STUDY DESIGN: 114 patients (52 females, 62 males) and 73 healthy controls (35 females, 38 males) aged 5 to 20 years were analysed for stage of puberty, age at menarche, testicular volume, basal serum follicle stimulating hormone (FSH), luteinising hormone (LH), sex hormone binding globulin (SHBG), testosterone and oestradiol concentrations, and free androgen index. RESULTS: Male gonadal and male and female pubic hair development occurred significantly earlier in the patients than in the controls. The mean age at menarche was significantly lower in the female patients than in their controls (11.7 v 13.2 years; p < 0.001), and lower than it had been for their mothers (v 13.1 years; p < 0.001). Relative testicular volume was higher in the male patients than in their controls (1.2 standard deviation score (SDS) v 0.2 SDS; p < 0.001). The prepubertal patients had higher basal LH (0.13 U/l v 0.08 U/l; p < 0.001) and SHBG (132.3 nmol/l v 109.1 nmol/l; p < 0.01) than the controls. Both the prepubertal and pubertal females had significantly higher basal FSH than their controls (1.57 U/l v 1.03 U/l; p < 0.05, and 4.0 U/l v 2.9 U/l; p < 0.01, respectively). CONCLUSIONS: Hydrocephalic children experience accelerated pubertal maturation, reflected in a younger age at menarche in females and an increased testicular volume in males. This may be because of enhanced gonadotrophin secretion, possibly resulting from unphysiological variations in intracranial pressure.     

Serum profiles of insulin-like growth factors and their binding proteins in adults with growth hormone receptor deficiency treated with insulin like growth factor I

Six adult patients with growth hormone receptor deficiency (GHRD) (2 men, 4 women) with an identical defect in the growth hormone receptor (GHR) gene, were treated with recombinant human insulin-like growth factor I (IGF-I), 40 μgikg S.C. twice daily, for 7 days. Serum concentrations of IGF peptide and IGF binding protein-3 (IGFBP-3) were measured by specific radioimmunoassays; serum IGFBPs were also measured by Western ligand blotting. The size distribution of both IGF-I and IGF-II was measured in serum following size-exclusion fast-performance liquid chromatography. IGF-I treatment resulted in a normalization of serum IGF-I levels on days 1–7 of treatment and a decrease in serum IGF-II levels. The fall in IGF-II levels and the simultaneous rise in IGF-I levels, however, resulted in an unchanged total serum IGF level. The low IGFBP-3 values did not significantly change during treatment, whereas there was a slight increase in IGFBP-2 levels. Preliminary analysis of size-fractionated sera suggested an increase in IGF-I levels in the 40 and 150 kDa regions at the expense of IGF-II levels. The results suggest that despite the failure of IGF-I treatment to increase IGFBPs significantly, serum IGFBP concentrations were sufficient to maintain normal levels of IGF-I. 0 Laron syndrome, growth hormone receptor deficiency, insulin-like growth factors, insulin-like growth factor binding protein     

Comparison of magnetometer and inductance plethysmography derived Konno-Mead diagrams during CO2 rebreathing

Magnetometers measure changes in antero-posterior diameters of the rib cage and abdomen while respiratory inductance plethysmography (RIP) measures changes in chest wall cross-sectional area. We compared Konno-Mead diagrams derived from isovolume calibrated magnetometers and RIP in the DC-mode during room air and CO2 rebreathing in the sitting and supine positions. Chest wall configurations obtained during quiet tidal breathing were similar in both sitting and supine positions. During CO2 stimulated breathing, however, derived chest wall configurations were at times dissimilar. The RIP per cent rib cage contribution to tidal volume was greater than that of magnetometers during room air and CO2 rebreathing in both sitting and supine positions. Changes in end expiratory levels measured by magnetometers and RIP during CO2 rebreathing are in general proportionate to each other; however, the magnetometers usually depicted quantitatively greater decreases in abdominal end expiratory levels during rebreathing. We conclude that the qualitative and quantitative assessment of chest wall configurations and volume displacement vary depending on the method used. RIP by including lateral wall motion may more accurately reflect compartmental displacements, while magnetometers by solely measuring antero-posterior diameter may better reflect changes in abdominal volume and thus diaphragm configuration.     

Molecular aspects of implantation: Blastocyst--endometrial interaction: an appraisal of some old and new ideas

The nature of the early interaction between blastocyst and endometrialepithelium is known to be highly specific within individualspecies. Despite this unique initial interaction, the trophoblastpromptly establishes a common theme as invasive cells penetratethe endometrium and colonize its local blood vessels. In allanimals with this type of implantation the blastocyst playsa more active role than the endometrial epithelium. The aggressivebehaviour of the blastocyst may be induced by signals from theendometrium which has been primed with preimplantation ovariansteroids. Activation of the blastocyst may reflect triggeringof synchronized paracrine loops, activated during the implantationwindow. Endometrial cytokines and eicosanoids are probably theprimary signals that drive the interlinking paracrine loopsand are essential for decidualization, trophoblast growth andinvasion. Since the dominant feature of early implantation israpid trophoblast migration, particularly in primates, the degreeto which the blastocyst attaches to the apical surface of theluminal endometrial epithelium during implantation is uncertain.The thickness of the glycocalyx on the uterine luminal epitheliumduring the peri-implantation period varies considerably betweenspecies. Its role in blastocyst attachment, if any, and in trophoblastcell locomotion, requires further study. The molecular propertiesand functions of the uterine epithelial plasma membrane, andthose of the interacting trophoblast at the site of implantation,have been largely neglected and require further extensive study. blastocyst adhesion/endometrial receptivity/implantation/plasma membrane/trophoblast     

Diaphragmatic EMG response to isocapnic hypoxia and hyperoxic hypercapnia in humans

The EMGdi response to both isocapnic hypoxia and hyperoxic hypercapnia was studied in the same sitting in six normal subjects. Rebreathing methods achieving "open loop" conditions were used. EMGdi was quantified as a moving time average. In almost all subjects, during hypoxia changes in EMGdi were inversely and hyperbolically related to changes in PAO2. When EMGdi was plotted against extrapolated O2 saturation, the relationship was linear in all subjects. The EMGdi response to hypoxia was qualitatively similar to the concurrent responses VI and P.15. EMGdi was linearly related to PACO2 during CO2 rebreathing. The slopes of the EMGdi response to decreasing O2 saturation were positively correlated to the slopes of the EMGdi response to PACO2, so that subjects with a low hypoxic response also had a low CO2 response and vice versa. The couplings of neural to muscular and muscular to ventilatory events as assessed by the ratio of the slopes of EMGdi to P.15 and P.15 to VI, respectively, were similar for all subjects and were not related to the degree or type of chemostimulation. The following were our conclusions. (1) EMGdi can be used as an index of respiratory motoneuron drive during hypoxic or hypercapnic breathing in normal humans. (2) The relative degree of responsiveness to hypoxic and hypercapnia stimuli (chemosensitivity) appears to be similar in any given individual. (3) In normal subjects, changes in inspiratory muscle pressure and ventilation are proportionate to changes in inspiratory neural drive as assessed by EMGdi.     

Effects of BCG Vaccination on Evolution of Leprosy

Background

BCG vaccine is considered to have immunoprophylactic potential in leprosy. However controversy exists about the extent of prophylaxis it provides. In view of this, a study was undertaken to see the evolution of disease in established cases of leprosy who were already vaccinated with BCG and to compare it with the cases of leprosy who were not vaccinated with BCG.

Methods

114 newly diagnosed cases of leprosy were studied. Patients were divided into two groups-BCG vaccinated and non-vaccinated. Clinical diagnosis of leprosy was confirmed by bacteriological and histopathological studies in each case. All patients were given standard anti leprosy treatment and were evaluated monthly for a minimum period of one year.

Results

All the cases were males in the age group of 20 to 50 years. 25.4% of cases had received BCG vaccine and 74.6% were not vaccinated for the same. No significant difference was observed in the incidence of different types of leprosy in vaccinated and non-vaccinated groups. 25.9% cases in non-vaccinated group developed lepra reaction as compared to 13.8% in vaccinated group. The incidence of deformities and disabilities in vaccinated group was only 10.3% as against 18.8% in non-vaccinated group. The rate of bacillary clearance appeared faster in vaccinated group.

Conclusions

Although there is no significant difference in the pattern of different types of leprosy in BCG vaccinated and non-vaccinated cases, there is reduction in the incidence of reactions as well as deformities and disabilities in BCG vaccinated cases as compared to non-vaccinated cases.Key Words: BCG Vaccine, Leprosy     

Environmental exposure characterization of fish processing workers

BACKGROUND: Aerosolization of seafood and subsequent inhalation, during processing is a potential high-risk activity for allergic respiratory disease. OBJECTIVES: To quantify total thoracic particulate, protein concentration and specific fish (pilchard, anchovy) antigen concentrations in fish processing plants; to determine the correlation between these exposure metrics; and to identify the major determinants of variability and the optimal grouping strategies for establishing dose-response relationships for fish antigen exposures. METHODS: Exposure assessments were conducted on randomly selected individuals within each of the identified 'exposure groups' (EGs) in two fish processing factories. Personal time-integrated sampling was conducted with a thoracic fraction sampler and analysed for particulate mass, total protein and specific fish antigens. Exposure metrics were developed on the basis of individually measured exposures and average levels of these personal samples within EGs. The main components of the exposure variability were determined using ANOVA techniques. RESULTS: A total of 198 full-shift personal aerosol samples were collected and analysed. Twenty-two percent of the samples were below the limit of detection (LOD) for pilchard and 23% for anchovy assays. Personal sampling revealed wide variations across EGs in arithmetic mean concentrations of thoracic particulate 0.61 mg m(-3) (range: LOD-11.3), total protein 0.89 microg m(-3) (LOD-11.5), pilchard antigen 150 ng m(-3) (LOD-15 973) and anchovy antigen 552 ng m(-3) (LOD-75 748) levels. The fishmeal loading and bagging sections of both plants showed consistently high thoracic particulate mass (0.811-2.714 mg m(-3)), total protein (0.185-1.855 microg m(-3)), pilchard antigen (538-3288 ng m(-3)) and anchovy antigen (1708-15 431 ng m(-3)). The a priori strategy that grouped workers according to EGs produced reasonably satisfactory summary exposure metric statistics. An alternative grouping strategy based on department revealed comparable elasticity (exposure contrast). While the correlation between the log-transformed thoracic particulate mass and fish antigen concentrations were generally modest (Pearson's r = 0.32-0.35, P < 0.001), a high correlation was found between pilchard and anchovy antigen concentrations (Pearson's r = 0.71, P < 0.001). Models using factory and department grouping strategies accounted for a significant portion of the variability (adjusted r(2) = 0.18, P = 0.043) in pilchard antigen levels. Grouping strategies using a combination of factory and department yielded the highest degree of elasticity for thoracic particulate (0.38) and pilchard antigen (0.42) levels. CONCLUSIONS: Workers involved in bony fish processing are at risk of inhaling aerosols containing pilchard and anchovy fish antigens. Antigen exposures are highest during fishmeal production and bagging. Grouping strategies based on department and factory may provide a more efficient approach than a priori classification of EGs for evaluating fish antigen exposures.