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Effect of medical and surgical therapy on aortic dissection evaluated by transesophageal echocardiography. Implications for prognosis and therapy. The European Cooperative Study Group on Echocardiography.

BACKGROUND: Aortic dissection still has a poor prognosis despite progress in therapy. Therefore, this prospective follow-up study was designed to determine whether the degree of communication between true and false lumen in relation to the type of dissection, analyzed by transesophageal echocardiography, influences the risk after initiation of medical or surgical therapy. METHODS AND RESULTS: In eight centers, 168 patients (124 men and 44 women) of age range of 23-84 years with proven aortic dissection were examined by transesophageal echocardiography in the acute phase, after start of medical and/or surgical therapy, and during follow-up (0-65 months; mean, 10 months). Analyses were performed prospectively according to a detailed study protocol. Patients were subdivided by transesophageal echocardiography according to a modified DeBakey classification. Type I aortic dissection was found in 35%, type II aortic dissection in 17%, and type III aortic dissection in 48%. Preoperative mortality was 3%, 7%, and 2%, and survival rates were 52%, 69%, and 70%, respectively. Type III aortic dissection could be subdivided into those with communication and antegrade dissection (ca) (50%), with communication and retrograde dissection limited to the descending aorta (cr desc) (10%), with dissection extended to the aortic arch and ascending aorta (cr asc) (27%), and with noncommunicating (nc) aortic dissection (13%). An open false lumen with no thrombus formation was present in types I, II, III ca and III cr asc aortic dissection in 17%, 21%, 39%, and 27% respectively, although it was most pronounced in types III nc and III cr desc (75% and 78%). During follow-up in patients who survived, thrombus was demonstrated in the false lumen in 80% of type I aortic dissection and 81% of types III ca and III cr asc. Open false lumen was seen in type II aortic dissection in 18%. Spontaneous healing was found in 4% with type II and 4% with type III aortic dissection (mainly in patients with type III nc aortic dissection). Patients with fluid extravasation, pleural effusion, pericardial tamponade, and periaortic effusion as well as mediastinal hematoma had a mortality of 52%. Reoperations were necessary in 12-29%, with the highest rate in patients with type III ca aortic dissection. Survival for patients with types III nc and III cr desc aortic dissection was higher than those with types I, II, III ca, and III cr asc. CONCLUSIONS: Preoperative mortality appears to be reduced by transesophageal echocardiography, allowing rapid initiation of treatment. Intraoperative and postoperative mortality in aortic dissection remains high. Risk factors are fluid extravasation and an open false lumen with high communication. Thrombus formation in the false lumen can be regarded as a good prognostic sign. Surgery appears to be only a first step in the treatment of aortic dissection. Second surgery or closure of entry sites based on intraoperative echocardiography may be considered to induce thrombus formation and reduce aortic wall stress.

Adult↗

Experimental pulmonary eosinophilia in mice by Ascaris suum extract.

The transnasal administration of an extract of the parasite Ascaris suum (Asc) to C57BL/6 mice for 3 wk produced marked eosinophilia in the bronchoalveolar lavage (BAL) fluid. This pulmonary eosinophilia was not accompanied by blood eosinophilia. The oral administration of cyclosporin, 50 mg/kg body weight, every other day significantly suppressed the pulmonary eosinophilia. Athymic C57BL/6-nu/nu mice failed to develop pulmonary eosinophilia. These data indicate that pulmonary eosinophilia caused by this parasite extract is T-cell-dependent. Genetically mast-cell-deficient (WB X C57BL/6)F1-W/WV (W/WV) mice developed marked eosinophilia in the BAL, which shows that mast cells are not necessary in the formation of eosinophilia in BAL in this model. C57BL/6, W/WV, and Balb-C mice that developed eosinophilia in the BAL also showed elevated total IgE levels and IgE titers against Asc in the sera. On the other hand, C57BL/6-nu/nu mice and IgE low-responder SJL mice, which developed little eosinophilia, did not show elevated total IgE levels and IgE titers against Asc in the sera. However, oral administration of cyclosporin, 50 mg/kg, which inhibited eosinophilia in the BAL in C57BL/6 mice, did not significantly inhibit the elevation of total IgE or IgE against Asc in the sera. This indicates that serum IgE production is not required for the formation of eosinophilia.

Animals↗

Aberrant lens fiber differentiation in anterior subcapsular cataract formation: a process dependent on reduced levels of Pax6.

PURPOSE: TGFbeta can induce development in lenses of opaque subcapsular fibrotic plaques that have many features of human subcapsular cataracts. To understand further the events associated with the onset and progression of TGFbeta-induced cataract, several different models for anterior subcapsular cataract (ASC) were used and characterized. METHODS: Anterior subcapsular plaques were induced in rat lenses cultured with TGFbeta and in transgenic mice overexpressing TGFbeta in the lens. ASC was also examined in lenses of mice haploinsufficient for Pax6, as well as in human biopsy specimens. Immunofluorescence and in situ hybridization labeling were used to examine changes in patterns of gene expression associated with cataract formation in these models. RESULTS: Examination of TGFbeta-induced cataract in transgenic mice established that the subcapsular plaques are composed of a heterogenous cell population: a population of myofibroblastic cells as well as a population of lens-fiber-like cells. Further support for phenotypic change comes from the observation that the cells in these plaques no longer expressed lens epithelial markers, such as Pax6 and Connexin43. Subsequent examination of human biopsy specimens of ASC, as well as lenses from Pax6-deficient mice, showed that the anterior subcapsular plaques in both cases were also composed of a heterogenous population of cells. In contrast, anterior subcapsular plaques that developed in vitro in response to TGFbeta did not have this same cellular heterogeneity, as no fiber-like cells were present. CONCLUSIONS: These findings suggest that in vivo, during TGFbeta-induced cataract formation, some lens epithelial cells transform into myofibroblastic cells, whereas others differentiate into fiber cells. As this pathologic change is accompanied by altered expression of genes characteristic of the normal lens epithelial cell phenotype and as lenses from Pax6-deficient mice exhibit development of anterior subcapsular plaques closely resembling those induced by TGFbeta in transgenic mice, the authors propose that a reduction in Pax6 levels may be essential for this pathologic process to progress. Furthermore, it is clear from these in vitro studies that TGFbeta alone cannot reproduce the same morphologic and molecular changes associated with ASC formation in vivo, indicating that additional molecule(s) in the eye are important in this process.

Actins↗

The effect of academic self-concept on ADHD and antisocial behaviors in early adolescence.

Using structural equation modeling techniques, we evaluated the effect of academic self-concept (ASC) on the development of attention-deficit/hyperactivity disorder (ADHD) and antisocial behaviors in early adolescence. Participants (n = 445) were recruited from the Dunedin Multidisciplinary Health and Development Research study. Eligibility was determined by the presence of complete data for the following variables at the specified time periods: reading at age 7, teacher reports of ADHD and antisocial behaviors at age 7, self-ratings of ASC at ages 9 and 11, and teacher reports of ADHD and antisocial behaviors at age 13. The results indicated that ASC is an important construct that directly contributes to the development of antisocial behaviors rather than to symptoms of ADHD. The results also indicated that children's early history of behavioral problems and academic performance contribute to the development of a more robust understanding of the impact of ASC on the development of disruptive behaviors in early adolescence.

Adolescent↗

Can semi-automated image cytometry on induced sputum become a screening tool for lung cancer? Evaluation of quantitative semi-automated sputum cytometry on radon- and uranium-exposed workers.

The correlations between semi-automated sputum cytometry (ASC), conventional cytology and the final diagnosis was investigated in industrially-exposed workers. Slides of sputum samples from 201 former uranium miners with silicosis, 100 patients with asbestosis, 103 workers resected for lung cancer, and 200 controls (50% smokers), were stained using the Papanicolaou (Pap) method and the Feulgen reaction with thionin. Cytometry was performed using the Cyto-Savant automated system. Atypical nuclei were found in 72 of 404 patient samples, 327 samples were normal and five were inadequate for ASC analysis. Thirteen tumours (Pap IV, Pap V) and 11 cases of severe dysplasia were identified by cytology. Lung cancer was confirmed in 20 patients. Compared to the final diagnosis of lung cancer, the sensitivity of ASC was 75% (15 out of 20) and specificity 89.8% (520 out of 579). The results represent a diagnostic efficiency of 89.3%. The combination of ASC with cytology increased sensitivity to 80% (16 out of 20) without significant loss of specificity (89.7% or 523 out of 581). In this investigation of a limited number of patients with occupational radon or asbestos exposure, semi-automated sputum cytometry appears to be sensitive and reliable for the detection of malignant changes in the tracheobronchial mucosa. Together with conventional cytology, it would be reasonable to test the validity of the combined methods in a large-scale feasibility study of early lung cancer detection.

Adult↗

Studies on collagen in the experimental myocardial infarction.

The healing process of myocardial infarction was studied with special reference to the production of collagen by the determinations of prolylhydroxylase (PHase) activity and contents of hydroxyproline (Hyp) of the collagen subfractions, i.e., neutral salt soluble (NSC), acid soluble (ASC) and insoluble collagens (ISC) in comparison with histologic observation of the proliferation of connective tissue in the canine heart. The contents of Hyp in NSC and ASC increased in the infarcted tissue 5 days after coronary ligation prior to its increase in ISC. The content of Hyp in NSC, which is a precursor of ASC and ISC, reached the maximal value at 7 days and that reached the maximal value in ASC 7 to 14 days after coronary ligation. ISC increased markedly after 14 days. PHase activity increased on the second day and attained to the highest at 5 to 7 days. Proliferation of fibroblasts paralleled with the increases in PHase activity and NSC, and an increase in collagen fiber did with the contents of ISC. From these results it is concluded that in the healing stage of myocardial infarction, PHase was activated earlier than the increase in immature collagen in the infarcted area which was observed 5 to 7 days after coronary ligation and was followed by the production of mature collagen after 14 days.

Animals↗

Chondrogenic and osteogenic differentiation of adipose-derived stem cells isolated from GFP transgenic mice.

Recent studies suggest that human adipose tissue contain pluripotent cells similar to bone marrow-derived stromal cells (BSCs). Taking advantage of homogeneously marked cells from green fluorescent protein (GFP) transgenic mice, we have previously demonstrated that BSCs differentiate into a variety of cell lineages both in vitro and in vivo). In the present study, we extend this approach to characterize adipose-derived stromal cells (ASCs)). These cells derived from human are sometimes called processed lipoaspirate (PLA) cells. ASCs were prepared from inguinal fat pads of GFP transgenic mice after extensive washing with PBS and treatment with collagenase. After the primary culture in control medium (DMEM + 10% FBS), the cells were incubated in either chondrogenic medium (DMEM + 1% FBS + insulin + ascorbate 2-phosphate + TGF-beta 1) or osteogenic medium (DMEM + 10%FBS + dexamethasone + ascorbate-2-phosphate + beta-glycerophosphate) for two to four weeks. Chondrogenic differentiation was assessed by Alcian blue staining, while osteogenic differentiation was by von Kossa and Alkaline phosphatase staining. ASCs incubated in chondrogenic medium induced Alcian blue positive cells. Incubation with osteogenic medium became positive for von Kossa and Alkaline phosphatase staining. No osteochondrogenic differentiation was observed in cells incubated with control medium. This cell population can be easily identified through fluorescence microscope, it should be an ideal source of ASCs for further experiments of stem cell biology and tissue engineering.

Adipocytes↗

Atypical glandular cells of undetermined significance. Outcome predictions based on human papillomavirus testing.

Cases of atypical glandular cells (AGC) diagnosed on liquid-based preparations were culled from a 3-year period. When available, residual cellular material was analyzed for human papillomavirus (HPV) by polymerase chain reaction and correlated with cytologic and histologic (biopsy) outcome. Of 178,994 cytologic cases, 187 (0.1045%) contained AGC compared with 8,740 (4.8828%) atypical squamous cells (ASC) for an AGC/ASC ratio of 0.021. HPV results and follow-up were available for 108 specimens from 106 patients. Depending on the end-point (histologic/cyto-logic), the sensitivity range of HPV testing for significant cervical disease (high-grade squamous intraepithelial lesion [SIL], adenocarcinoma in situ [ACIS], invasive carcinoma) was 83% with a specificity range of 78% to 82%, a positive predictive value of 57% to 61%, and a negative predictive value of 91% to 95%. Fifteen false-positive results included concurrent ASC or low-grade SIL, ASC on follow-up cytology, and previous ACIS with a negative follow-up cone biopsy result. Noncervical glandular neoplasia (including atypical endometrial hyperplasia) was confirmed in 13 cases (1 recurrent), only 2 of which scored positive for HPV. HPV-positive AGC has a substantially higher positive predictive value for significant disease than ASC (61% vs historic 20%) and merits consideration in the triage of patients with atypical endocervical cells not otherwise specified. However, noncervical or other HPV-negative glandular neoplasia must be considered in all patients with AGC, particularly older patients.

DNA, Viral↗

Atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion. A follow-up study of conventional and liquid-based preparations in a high-risk population.

We compared the histologic follow-up of 368 smears or slides with an interpretation of "atypical squamous cells, cannot rule out high-grade squamous intraepithelial lesion" (ASC-H) based on conventional and liquid-based preparations and age groups in a high-risk population. Patients with an ASC-H interpretation were 17 to 87 years old (mean, 36.8 years). The specimens were 52 liquid-based preparations and 316 conventional smears. For 218 cases (59.2%), including 28 liquid-based preparations (65%) and 190 conventional smears (58%), histologic follow-up was available. In 20 liquid-based preparations (71%) and 152 conventional smears (80.0%), cervical intraepithelial neoplasia (CIN) or higher was revealed on subsequent biopsy. Other results were as follows: liquid-based preparations, CIN1, 11 (55%); CIN2/3, 9 (45%); conventional smears, CIN1, 78 (51.3%); CIN2/3, 70 (46.1%); squamous cell carcinoma, 4 (2.6%). There was no statistically significant difference in the incidence of CIN or higher on subsequent biopsy after an interpretation of ASC-H based on preparation types. The incidences of CIN in patients 40 years old or older and patients younger than 40 years were 66% and 84%, respectively, a statistically significant difference. Because of the high incidence of clinically significant lesions noted on subsequent follow-up, patients with an interpretation of ASC-H should be observed closely and referred for colposcopic examination regardless of their age.

Adolescent↗

Impaired expression of noncollagenous bone matrix protein mRNAs during fracture healing in ascorbic acid-deficient rats.

In scorbutic patients, fractures are slow to heal because of impaired collagen synthesis. To investigate the influence of impaired collagen synthesis on the differentiation and proliferation of osteogenic and chondrogenic cells, we examined the expression of genes encoding bone matrix proteins, including osteonectin (ON), osteopontin (OPN), osteocalcin (OC), and matrix Gla protein (MGP), as differentiation markers for osteogenic and chondrogenic cells during fracture healing in Osteogenic Disorder Shionogi (ODS) rats, which have a hereditary defect in the ability to synthesize ascorbic acid (Asc). In ODS rats without Asc supplementation, intramembranous ossification was completely inhibited. Although a few fibroblast-like cells expressing ON mRNA were observed, no OPN mRNA-expressing cells were detected. During endochondral ossification, a small amount of metachromatic staining cartilage appeared at the fracture site, but there was no provisional calcification zone in the cartilage. Chondrocytes expressed ON and MGP mRNAs, but not OPN mRNA. When Asc was given to these rats, callus formation was soon detected around the fracture site, while OPN mRNA was expressed by differentiated osteoblasts and hypertrophic chondrocytes. Our data indicate that impaired collagen synthesis due to Asc deficiency inhibited the increase of ON and MGP mRNA-expressing cells as well as the appearance of OPN mRNA-expressing cells. Since OPN is considered to play an important role in normal and pathological mineralization, lack of OPN mRNA expression accompanying impaired collagen synthesis may have a role in defective mineralization and delayed fracture healing in scurvy.

Animals↗

Screening for cervical cancer: new alternatives and research.

Evidence for the clinical utility of human papillomavirus (HPV) DNA testing has increased over the years and has now become very convincing. Some specific uses of HPV detection are a) triage of women with cytological determinations of atypical squamous cells of undetermined significance (ASC-US) and related management strategies, b) as a marker for test of cure post-treatment, and c) most importantly, as an adjunct to cytology in routine cervical disease screening programs. There are many studies that support each of these applications and include 8 studies on ASC-US triage, 10 on test of cure and 13 on adjunctive or stand-alone HPV screening. The most notable investigation of ASC-US triage was ALTS, a randomized controlled trial of 3,488 women. With respect to routine HPV screening the combined studies included 77,000 women, providing as a histological endpoint more than 1,000 cases of high-grade cervical intraepithelial neoplasia (CIN) or cancer. Testing methods were either the Hybrid Capture 2 (HC2) test or the polymerase chain reaction (PCR) test. HPV testing of women with ASC-US cytology had on average a higher sensitivity (90%) and specificity (70%) than repeating the cytological test (sensitivity 75%, specificity 60%) and was also more sensitive than colposcopy for follow-up. As an adjunct to the Papanicolaou (Pap) cytology test in routine screening, HPV DNA testing was a more sensitive indicator for prevalent high-grade CIN than either conventional or liquid cytology. A combination of HPV DNA and Papanicolaou testing had almost 100% sensitivity and negative predictive value. The specificity of the combined tests was slightly lower than the specificity of the Papanicolaou test. One "double-negative" HPV DNA and Papanicolaou test indicated a higher prognostic assurance against risk of future CIN 3 than three subsequent negative conventional Papanicolaou tests and may safely allow three-year or longer screening intervals for such low-risk women. It appears that HPV DNA testing is on the way to becoming a common testing strategy in cervical cancer prevention programs. Research continues into approaches for improving the performance and cost-effectiveness of HPV detection methods. Hybrid Capture 3 will offer improved HPV typing capabilities and the Rapid Capture machine allows for robot-assisted HPV DNA testing, permitting greater test throughput. PCR test improvements are expected to contribute to the growth of flexible accurate and cost-effective HPV DNA tests. It is likely that improved diagnostic technology along with HPV genotyping and quantitation may provide more value in future. A particularly promising approach is to combine HPV DNA testing with expression levels of other markers such as proliferative or cell cycle regulatory proteins to subdivide HPV-positive women into those who are at greater risk of cancer and those who can be safely followed by screening at longer intervals. This paper is available too at: http://www.insp.mx/salud/index.html.

Biomarkers↗

[The relationship between the intracellular redox status of immune cells and progression of hepatitis C virus related chronic liver disease].

It has been suggested that oxidative stress participates in the pathogenesis of hepatitis C virus infection. It also has been made clear that redox status in T cell and macrophage relates to the activity of virus infectious disease such as HIV infection. With such background we evaluated the relationship between the intracellular redox status of T cell and macrophage and the activity of HCV positive chronic liver disease. Intracellular GSH and GSSG levels of T cell and macrophage were determined in twenty-five HCV positive asymptomatic carriers (C-ASC), sixty-three chronic hepatitis patients (C-CH), ten HCV positive liver cirrhosis patients (C-LC) and twenty-nine healthy controls. The intracellular GSH levels of T cell (T-GSH) significantly decreased in both C-CH and C-LC compared with healthy controls. No significant differences in the T-GSH levels were found between healthy controls and C-ASC. T-GSH levels of C-CH and C-LC were significantly lower compared with C-ASC. The intracellular GSH levels of macrophage (CD14-GSH) of C-LC were significantly decreased compared with healthy controls. The CD14-GSH levels of C-CH and C-LC were significantly lower compared with C-ASC. There was no correlation between intracellular GSH, GSSG levels and the serum levels of iron-related markers, fibrogenesis markers and other clinical parameters. These results suggest that the intracellular redox status of T cell and macrophage relates to the progression of HCV related chronic liver disease.

Aged↗

L-ascorbic acid 2-phosphate promotes osteoblastic differentiation of MC3T3-E1 mediated by accumulation of type I collagen.

L-Ascorbic acid 2-phosphate (Asc-P: a stable ascorbic acid derivative) markedly stimulated synthesis of marker proteins for osteoblastic differentiation such as alkaline phosphatase and osteocalcin in a murine osteoblastic cell line, MC3T3-E1, suggesting that Asc-P could promote osteoblastic differentiation. L-Azetidine 2-carboxylate (AzC) diminished the stimulatory effects of Asc-P on the synthesis of alkaline phosphatase and osteocalcin due to its inhibitory effects on mature collagen secretion. Growing cells on the dishes coated with type I collagen resulted in an increased expression of osteoblastic phenotypes even in the presence of AzC. Coating with fibronectin, however, failed to promote differentiation. These results suggest that the promotion of cell differentiation caused by Asc-P is mediated by the accumulation of mature collagen.

Alkaline Phosphatase↗

Effect of simulated spray chilling with chemical solutions on acid-habituated and non-acid-habituated Escherichia coli O157:H7 cells attached to beef carcass tissue.

Samples (10 by 20 by 2.5 cm) of beef carcass tissue were inoculated (10(4) to 10(5) CFU/cm2) with Escherichia coli O157: H7 that was either non-acid habituated (prepared by incubating at 15 degrees C for 48 h in inoculated filter-sterilized composite [1:1] of hot and cold water meat decontamination runoff fluids, pH 6.05) or acid habituated (prepared in inoculated water fluids mixed with filter-sterilized 2% lactic acid [LA] runoff fluids in a proportion of 1/99 [vol/vol], pH 4.12). The inoculated surfaces were exposed to conditions simulating carcass chilling (- 3 degrees C for 10 h followed by 38 h at 1 degree C). Treatments applied to samples (between 0 and 10 h) during chilling included the following: (i) no spraying (NT) or spraying (for 30 s every 30 min) with (ii) water, (iii) cetylpyridinium chloride (CPC; 0.1 or 0.5%), (iv) ammonium hydroxide (AH; 0.05%), (v) lactic acid (LA; 2%), (vi) acidified sodium chlorite (ASC; 0.12%), (vii) peroxyacetic acid (PAA; 0.02%), (viii) sodium hydroxide (SH; 0.01%), or (ix) sodium hypochlorite (SC; 0.005%) solutions of 4 degrees C. Samples were taken at 0, 10, 24, 36, and 48 h of the chilling process to determine changes in E. coli O157:H7 populations. Phase 1 tested water, SH, PAA, LA, and 0.5% CPC on meat inoculated with non-acid-habituated pathogen populations, whereas phase 2 tested water, SC, AH, ASC, LA, and 0.1% CPC on meat inoculated with acid- and non-acid-habituated populations. Reductions in non-acid-habituated E. coli O157:H7 populations from phase 1 increased in the order NT = water = SH < PAA < LA < CPC. Reductions from phase 2 for acid-habituated cells increased in the order NT = water = SC < ASC = LA = AH < CPC, whereas on non-acid-habituated cells the order observed was NT = water = SC < AH = ASC < LA < CPC. Previous acid habituation of E. coli O157:H7 inocula rendered the cells more resistant to the effects of spray chilling, especially with acid; however, the trend of reduction remained spray chilling with water = non-spray chilling < spray chilling with chemical solutions.

Animals↗

Comparison of human papillomavirus DNA prevalence in atypical squamous cells of undetermined significance subcategories as defined by the original Bethesda 1991 and the new Bethesda 2001 Systems.

CONTEXT: The new Bethesda System 2001 (TBS 2001) minimized the subclassification of atypical squamous cells of undetermined significance (ASCUS). OBJECTIVE: The primary goal of this study was to determine the impact of the new subclassification on the accuracy of Papanicolaou (Pap) test diagnosis by examining the prevalence of human papillomavirus (HPV) DNA in different ASCUS subcategories, as defined by the new TBS 2001 versus the original TBS 1991. The second goal was to identify specific morphologic features of atypical squamous cells that are more frequently associated with HPV detection. DESIGN: Consecutive cases of ThinPrep Pap tests were retrospectively reviewed by a panel of pathologists to obtain consensus diagnoses. The study group consisted of ASCUS cases; the positive control group consisted of low- and high-grade squamous intraepithelial lesions (LSILs and HSILs, respectively); and the negative control group consisted of cases "negative for intraepithelial lesion or malignancy." All ASCUS cases were subclassified according to TBS 1991 into the following categories: favor reactive (ASCUS-R), favor LSIL (ASCUS-L), favor HSIL (ASCUS-H), and not otherwise specified (ASCUS-NOS). In a separate review, ASCUS cases were subclassified according to TBS 2001 into the following categories: atypical squamous cells of undetermined significance (ASC-US) and atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion (ASC-H). Furthermore, morphologic ASCUS subtypes were recorded (atypical mature, immature, parakeratotic, and atrophic cells); in addition, individual morphologic features of atypical cells were recorded. Broad- spectrum HPV DNA amplification and genotyping was performed using short PCR fragment (SPF 10) polymerase chain reaction/Line Probe assays. RESULTS: In cases classified according to TBS 1991, HPV was detected in 32% of negative, 49% of ASCUS, and 93% of LSIL/HSIL cases. On the second review, using the diagnostic categories of TBS 2001, which eliminated the ASCUS-;R category, the number of ASCUS cases decreased by 45%. The prevalence of HPV DNA in ASCUS cases downgraded to the negative category was 38%, which was not significantly different from HPV prevalence in negative cases as diagnosed under TBS 1991. Furthermore, HPV was detected in 56% of ASC-US and 71% of ASC-H cases. The prevalence of HPV in different morphologic subtypes of ASCUS was not significantly different, and none of the 8 individual morphologic features of atypical cells were more frequently associated with HPV detection. CONCLUSION: Elimination of the ASCUS-R category in TBS 2001 resulted in a significant decrease in the number of ASCUS diagnoses. Downgraded cases had a relatively low prevalence of HPV DNA. It is expected that TBS 2001 will increase specificity of the Pap test without compromising its sensitivity.

Adolescent↗

A drug distribution system to supply a detached ambulatory surgery center from the main hospital's central pharmacy.

Ambulatory surgery centers (ASCs) and other outpatient service facilities are becoming more prevalent throughout the country. The development and implementation of a drug distribution system to supply a detached ambulatory surgery center is described. The method proposed to provide all the medication needed for the ASC uses a unit-dose cart with lockable doors. The cart is delivered to the ASC each morning by nonpharmacy hospital transportation personnel. A locked case method similar to the system described by McClure-Zola et al. is employed for schedule II controlled substances. Specific formularies were developed for the anesthesia and the operating/recovery rooms. Medications for both areas are arranged separately in the cart. The cart is returned to the pharmacy at the end of each ASC day to be replenished on the night shift by the pharmacy staff. Additional full-time equivalents are not required in the pharmacy budget to provide this service. The system described provides a simple method for efficiently and effectively meeting the medications needs of a free-standing satellite of the hospital.

Centralized Hospital Services↗

[Presence of alpha 5 beta 1 integrin and fibronectin in the anterior subcapsular cataract].

PURPOSE: We investigated whether alpha 5 beta 1 integrin and fibronectin were present in myofibroblast-like lens epithelial cells in anterior subcapsular cataract (ASC). METHODS: Nine anterior capsule specimens were obtained from the patients during cataract surgery and frozen for cryostat sections. Six specimens were anterior capsule obtained from cataract with ASC. As a control, three specimens were obtained from cataract without ASC. Alpha-smooth muscle actin (alpha-SMA), alpha 5 beta 1 integrin, and fibronectin were detected by immunohistochemical observation. RESULTS: In all 6 specimens from patients with ASC, the lens epithelial cells around fibrosis tissue included myofibroblast-like lens epithelial cells which were positive for alpha-SMA. alpha 5 beta 1 integrin was detected in these lens epithelial cells. Fibronectin was also detected around these myofibroblast-like lens epithelial cells. Three control specimens showed no immunoreactivity against alpha-SMA, alpha 5 beta 1 integrin, or fibronectin. CONCLUSIONS: Alpha 5 beta 1 integrin and fibronectin may play an important role in myodifferentiation of lens epithelial cells.

Adult↗

Biochemical characterization of the soluble form of tumor antigen MUC1 isolated from sera and ascites fluid of breast and pancreatic cancer patients.

Transmembrane glycoprotein tumor antigen MUC1 that is overexpressed on pancreatic and breast tumor cells can be found in large amounts in soluble form in serum and ascites fluid. MUC1 has been identified as a target of human antitumor antibody and CTL responses that are generated in the absence of helper T cells. The soluble form of MUC1 should support generation of helper T cells, but we have found recently that this form, although effectively endocytosed by dendritic cells, remains trapped in early endosomes and is not trafficked to antigen-processing compartments. The exact biochemical structure of this form of MUC1 has not been elucidated to date, and it is thus not clear what structural characteristics may be responsible for its retention in early endosomes. We have purified soluble MUC1 from ascites fluid of breast/pancreatic cancer patients (ASC-MUC1) and quantitated O-linked carbohydrates. We have altered ASC-MUC1 by enzymatic treatment: trypsin or clostripain digestion, desialylation, and further in vitro glycosylation. We have found that desialylated ASC-MUC1 was further glycosylated by peptidyl N-acetylgalactosamine transferases and was not when sialic acid was present. These alterations created new forms of ASC-MUC1 that might be handled more efficiently by antigen-presenting cells to generate better tumor-specific immunity and used to identify structures that are directly involved in retention of this antigen in early endosomes.

Ascites↗