Multiparametric deoxyribonucleic acid and cell cycle analysis of breast carcinomas by flow cytometry. Clinicopathologic correlations.

Using intact ethanol-fixed cytokeratin monoclonal (CAM 5.2) and propidium iodide dual-stained cells, we have performed two-color multiparametric flow cytometric (FCM) DNA analysis and S-phase fraction (SPF) determination on 165 mechanically dissociated breast carcinomas. Sixty-seven patients were axillary node positive, 33 patients node negative; 59 had biopsy only and in 8, FCM was performed on tissue from metastatic lesions. Overall, 62% of the tumors contained aneuploid cell populations. Abnormal cellular DNA content (aneuploidy) was significantly correlated with high nuclear grade (p less than 0.001), lack of estrogen receptors (p less than 0.001), presence of vascular invasion (p less than 0.04), high histologic grade (p less than 0.04), and tumor size (p less than 0.03) but not with patient age (p greater than 0.07) or axillary node status (p greater than 0.50). SPF values derived from ungated histograms had a positively skewed frequency distribution (range 2 to 30%, N = 152) with an overall median of 11% (diploid, 8.9%; aneuploid, 15.7%). Higher SPF values were significantly correlated with aneuploidy (p less than 0.001), presence of necrosis (p less than 0.001), lack of estrogen receptor (p less than 0.0001), high nuclear grade (p less than 0.001), vascular invasion (p less than 0.003), tumor size (p less than 0.006), and high histologic grade (p less than .004) but not the presence of lymph node metastases (p greater than 0.56). Mean SPF values were significantly higher when calculated from cytokeratin gated DNA histograms (14.1% versus 11.5%, p less than 0.001), probably due to exclusion of contaminating stromal/inflammatory cells; and significantly lower when calculated from debris subtracted histograms (7.8% versus 11.4%). Cytokeratin gated and debris subtracted SPF values both had a greater degree of correlation than ungated values with clinicopathologic factors of known prognostic significance.(ABSTRACT TRUNCATED AT 250 WORDS)     

Identification ofSchistosoma haematobium soluble egg antigens that elicit human granuloma formation in vitro

Schistosoma haematobium soluble egg antigens (SH SEAs) induce intense granulomas in human hosts that often culminate in severe disease. In an attempt to identify the SH SEA fractions that are responsible for pathology, we combined T-cell Western blotting and an in vitro model of granuloma formation. Whole SH SEAs were dotted onto nitrocellulose pieces or were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrotransferred onto nitrocellulose paper. Horizontal strips bearing the separated antigens were solubilized in dimethylsulfoxide and precipitated in carbonate/bicarbonate buffer. Antigen-free and antigen-bearing particles were used to stimulate peripheral blood mononuclear cells (PBMCs) obtained fromS. haematobium-infected patients and sex- and agematched healthy controls to form granulomas in vitro. Whole SH SEA-bearing nitrocellulose particles elicited in vitro formation of granulomas by PBMCs from infected donors. The response was similar in sensitivity, specificity, and reproducibility to that evoked by SH SEA-bound polyacrylamide beads. The results obtained in samples from 30 patients and 10 controls tested with SH SEA-separated fractions revealed that SEA bands of 84 000, 63 000, 57 000, 55 000, 40 000, 30 000, and 28 000 Da elicited in vitro granuloma reactions by PBMCs of almost all infected patients. Conversely, separated soluble adult-worm antigens failed to stimulate PBMCs of infected patients to form granulomas. This study is the first to identify the SH SEA fractions that evok in vitro granuloma formation and represents an initial step toward the development of an anti-urinary schistosomiasis pathology vaccine.     

Cytogenetics of primary prostatic adenocarcinoma. Clonality and chromosome instability.

We have examined 62 prostatic adenocarcinomas by conventional cytogenetic analysis. Most were primary cultures harvested in 14 days or less. The most consistent finding was a normal male diploid karyotype, found in 87% of all cells analyzed, and as the exclusive finding in 19 tumors. Nonrandom chromosomal changes included gain of chromosome 7 and loss of the Y chromosome. In addition, clonal gains of chromosomes 8, 12, and 18, and clonal losses of chromosomes 14 and 19 were noted in individual cases. Two structural clonal aberrations, a 9p+ in one case and a t(Y;22) (q11.2;p12) in another, were also seen. Ten of 62 cultures demonstrated chromosome instability, defined herein as nonclonal gain or loss of chromosomes in more than 10% of the metaphases examined from that culture. In those cases with nonclonal numerical aberrations, loss of chromosomes was more common than gain. The distribution of apparently random numeric abnormalities was similar to that of the clonal abnormalities in that the most frequent nonclonal gain was of chromosome 7 and the most frequent nonclonal loss was of the Y chromosome. Apparently random structural aberrations were observed in less than 1% of all analyzed cells. These included a 4p-,del(3)(q13), and t(1;11). The extent of apparently random aneuploidy suggests that chromosome instability characterizes cultured prostatic adenocarcinomas. An increase in the frequency of nonclonal aberrations may be an indicator of tumor origin in a predominantly diploid cell population. The coexistence of clonally aberrant, nonclonally aberrant, and normal diploid cells in culture may reflect heterogeneity of prostate tumors in vivo.     

Survivorship and Patient-Reported Outcomes of an Uncemented Vitamin E–Infused Monoblock Acetabular Cup: A Multicenter Prospective Cohort Study

BackgroundAddition of vitamin E to polyethylene is theorized to reduce the potential for oxidative wear in acetabular components. This paper presents a multicenter prospective cohort study that reports on outcomes from use of a Vitamin E–infused highly cross-linked polyethylene acetabular cup.MethodsPatients were recruited across nine medical institutions. Clinical outcome measures recorded were the Harris Hip Score, visual analogue score for pain and satisfaction. Evidence of implant loosening or osteolysis was collected radiologically. Cup survival and reasons for revision in relevant cases were also recorded. Data collection was undertaken preoperatively, at 6-12 weeks, 6 months, 1 year, 2 years, and 5 years. A total of 675 patients were recruited, with 450 cases available at final review. Data regarding cup survival was available to 8 years and 9 months postoperatively.ResultsImprovements in both the Harris Hip Score and visual analogue score for pain and satisfaction were recorded at all time points, with these being maintained through the length of follow-up. In total, 89% of cups were implanted within the Lewinnek safe zone. A lucent line was identified in one case, with no evidence of acetabular osteolysis observed throughout the follow-up period. Cup survival was 98.9% at 8 years and 9 months. No revisions for aseptic loosening were observed.ConclusionsThe use of a vitamin E–infused polyethylene acetabular cup demonstrates reassuring patient-reported outcomes, radiological measures, and cup survival at medium to long-term follow-up.     

Outcome of lateral pelvic lymph node dissection with total mesorectal excision in treatment of rectal cancer: A systematic review and meta-analysis

BackgroundTotal mesorectal excision is the gold standard treatment of mid- and low-lying rectal cancer. Lateral pelvic lymph node dissection has been suggested as an approach to decrease recurrence and improve survival. Our meta-analysis presented here aimed to review the current outcomes of lateral pelvic lymph node dissection and total mesorectal excision in comparison with total mesorectal excision alone.MethodsA systematic literature search querying electronic databases was conducted in accordance with the Preferred Reporting Items for Systematic reviews and Meta-Analyses guidelines. We reviewed articles that reported the outcomes of lateral pelvic lymph node dissection combined with total mesorectal excision in comparison with total mesorectal excision alone. The main outcome measures were local recurrence, distant metastasis, overall and disease free-survival, and complications.ResultsThis systematic review included 29 studies of 10,646 patients. Of those patients, 39.4% underwent total mesorectal excision with lateral pelvic lymph node dissection. The median operation time for the lateral pelvic lymph node dissection + total mesorectal excision was significantly longer than total mesorectal excision alone (360 minutes versus 294.7 minutes, P = .02). Lateral pelvic lymph node dissection + total mesorectal excision was associated with higher odds of overall complications (odds ratio = 1.48, 95% confidence interval: 1.18–1.87, P < .001) and urinary dysfunction (odds ratio = 2.1, 95% confidence interval: 1.21–3.67, P = .008) than total mesorectal excision alone. Both groups had similar rates of male sexual dysfunction (odds ratio = 1.62, 95% confidence interval: 0.94–2.79, P = .08), anastomotic leakage (odds ratio = 1.15, 95% confidence interval: 0.69–1.93, P = .59), local recurrence (hazard ratio = 0.96, 95% confidence interval: 0.75–1.25, P = .79), distant metastasis (hazard ratio = 0.96, 95% confidence interval: 0.76–1.2, P = .72), overall survival (hazard ratio = 1.056, 95% confidence interval: 0.98–1.13, P = .13), and disease-free survival (hazard ratio = 1.02, 95% confidence interval: 0.97–1.07, P = .37).ConclusionLateral pelvic lymph node dissection was not associated with a significant reduction of recurrence rates or improvement in survival as compared with total mesorectal excision alone; however, LPLND was associated with longer operation time and increased complication rate.     

Nuclear morphometry for predicting cervical metastatic potential in laryngeal squamous-cell carcinoma

Quantitative nuclear morphometry (QNM) provides a reliable, reproducible method for measuring the degree of nuclear pleomorphism, a qualitative parameter widely incorporated into histopathological grading schemes. Nuclear shape has successfully been shown to predict primary prostate adenocarcinoma metastatic potential. This investigation utilized QNM to measure the nuclear pleomorphism of 50 patients with primary laryngeal squamous cell carcinoma to determine whether this computer-assisted method could discriminate between those patients with and without cervical metastases. Nuclear cross-sectional area and nuclear roundness as determined by QNM could not discriminate between patients with and without cervical metastases, though significant alterations in nuclear morphology exist between normal laryngeal squamous epithelium and laryngeal squamous cell carcinoma.     

Emergence of Salmonella enteritidis outbreaks in broiler chickens in the Lebanon: epidemiological markers and competitive exclusion control

This study investigates the first emergence of Salmonella Enteritidis outbreaks among chickens in the Lebanon and identifies the epidemiological markers of selected recovered Enteritidis strains. In addition, the authors evaluate a competitive exclusion approach to control infection in broiler chickens by Enteritidis organisms which possess the prevalent identified markers. The basic procedure in this investigation involved recording signs and lesions in eleven broiler chicken flocks on eleven farms, and culturing livers, spleens, and caeca of ten randomly selected birds per flock for Salmonella isolation and serotyping. Furthermore, culturing for Salmonella and serotyping was attempted from the livers, spleens, caeca and oviduct swabs of ten hens in four broiler breeder flocks which provided hatching eggs for the broilers under study. The identification of epidemiological markers in recovered S. Enteritidis included the determination of drug-resistance patterns and plasmid profiling. The competitive exclusion was evaluated by spraying the microflora on day-old broilers in the hatchery, followed by a controlled oral challenge at three days of age, with 2.85 x 10(5) colony-forming units of S. Enteritidis organisms per bird. Exclusion was evaluated by culturing for S. Enteritidis in anal swabs, spleens, livers, and caeca of individual challenged birds treated with the microflora and in untreated challenged birds. A total of 112 invasive S. Enteritidis strains were recovered on eleven farms from individual organs of broiler chickens with typical signs and lesions of salmonellosis. The prevalent resistance to drugs in such strains was to furaltadone and gentamycin, a marker identified in 93 strains (83%), recovered from nine out of eleven farms. The same resistance pattern was present in S. Enteritidis strains recovered from breeders on one out of four farms. The prevalent plasmid profile in nine S. Enteritidis organisms selected randomly from a pool of 93 strains (one per each of the nine broiler farms) was 14.1 kilobases (kb) and approximately 50.0 kb, a typical pattern to that identified in S. Enteritidis organisms recovered from oviducts of breeders on one out of four breeder farms. The exclusion significantly reduced cumulative mortality in birds of up to 45 days of age by 3.93%, in comparison to that observed in untreated challenged birds (P < 0.05). At 45 days of age, exclusion resulted in a 15.6% reduction in the percentage infection rate by S. Enteritidis in spleens or livers and a 34.4% reduction in the percentage infection rate of the caeca (P < 0.05).