P53 mutation analysis of colorectal liver metastases: relation to actual survival, angiogenic status, and p53 overexpression.

PURPOSE: To correlate TP53 mutations with angiogenic status of the tumor and prognosis after liver surgery in patients with colorectal liver metastases and to correlate immunohistochemical staining of p53 protein with TP53 gene mutations. EXPERIMENTAL DESIGN: Tumors of 44 patients with surgically treated colorectal liver metastases were analyzed for (a) TP53 mutations using denaturing gradient gel electrophoresis followed by sequencing, (b) microvessel density using the hot spot overlap technique, (c) apoptotic rate in tumor cells and endothelial cells of tumor microvessels using double immunostaining for anti-cleaved caspase 3 and anti-CD34, and (d) expression of p53 protein using immunohistochemistry. RESULTS: TP53 mutations were detected in 36% of the metastases and occurred more frequently in liver metastases from left-sided colon tumors than from right-sided colon tumors (P = 0.04). In metastases with TP53 mutations, microvessel density was higher compared with tumors with wild-type p53. Endothelial cell apoptosis was not different in tumor microvessels from TP53-mutated versus nonmutated tumors. The 5-year actual survival was not influenced by TP53 mutational status, microvessel density, or endothelial cell apoptotic rate of the tumors. Based on immunohistochemical p53 overexpression, the positive and negative predictive values of TP53 mutations were 61% and 82%. CONCLUSIONS: In patients with surgically treated colorectal liver metastases, TP53 mutations and angiogenic status did not influence prognosis. Immunohistochemistry is not a reliable technique for detecting TP53 mutations.     

Long-term NaHS administration reduces oxidative stress and apoptosis in a rat model of left-side varicocele

The main aim of this study was to assay the testicular H₂S levels in the varicocele rat model and then to investigate the protective effects of NaHS on morphometric changes, sperm parameters, oxidative stress and apoptosis markers in rat's testis. D,L-propargylglycine (PAG) was administrated to show the effects of cystathionine γ-lyase enzyme (CSE) inhibition in the varicocele. Rats were assigned to four groups: (a) Sham, (b) varicocele, (c) varicocele + PAG and (d) varicocele + NaHS. Animals in varicocele + NaHS group received 30 µmol/L NaHS in drinking water for 56 days. In the varicocele + PAG group, animals received PAG 19 mg/kg twice a week. Morphometric assessment, oxidative stress markers, testicular H₂S levels, sperm parameters, TUNEL assay and expression of Bax/Bcl2 were evaluated at the end of experiment. Testicular H₂S levels were significantly decreased in varicocele group. NaHS significantly improved sperm parameters, morphometric characteristics and oxidative stress compared to varicocele group. Oxidative stress status deteriorated in the PAG group compared to the varicocele group. This study showed that a low testicular H₂S level might play a critical role in male infertility. Thus, NaHS administration may be a promising treatment strategy for male infertility in varicocele. In addition, CSE may not be the only important enzyme in testicular H₂S production.     

Clinical Validation of the Cervista HPV HR Test According to the International Guidelines for Human Papillomavirus Test Requirements for Cervical Cancer Screening

This study demonstrates that both the clinical sensitivity and specificity of the Cervista HPV HR test for high-risk human papillomavirus (HPV) detection are not inferior to those of the Hybrid Capture 2 (HC2) test. The intra- and interlaboratory reproducibilities of Cervista were 92.0% (kappa, 0.83) and 90.4% (kappa, 0.80), respectively. The Cervista HPV HR test fulfills all the international HPV test requirements for cervical primary screening purposes.     

Lymphvascular space involvement compromises the survival of patients with stage I endometrial cancer: results of a multicenter study.

AIMS: To quantify the relative risk associated with lymphvascular space involvement (LVSI) on outcome measures in patients with apparent stage I endometrial cancer. METHODS: Six hundred and ninety nine consecutive patients with endometrial carcinoma apparent stage I, who underwent surgery in one of four gynecological oncology centers in Israel, comprised the study population. Forty cases with and 659 without LVSI were followed for a median time of 39 months. Recurrence free, disease specific and overall survival was compared between the two groups. The effect of LVSI, adjusted for other clinical and histo-pathological prognostic factors, was assessed by multivariate analysis. RESULTS: The univariate Kaplan-Meier procedure for survival analysis showed that patients with LVSI had lower recurrence free survival (p=0.0003), worse disease specific (p=0.0007) and overall survival (p<0.0001). Cox proportional hazards model demonstrated a trend toward shorter recurrence free survival (HR=2.0, 95% CI 0.9, 4.5; p=0.08), a worse disease specific survival (HR=2.8, 95% CI 1.1, 7.4; p=0.04) and decreased overall survival (HR=2.0, 95% CI 1.1, 3.8; p=0.03) in cases with LVSI. CONCLUSIONS: In patients with apparent stage I endometrial cancer the presence of LVSI, an independent poor prognostic factor, is associated with a two fold increased risk of death. The presence of LVSI warrants consideration when deciding upon post operative management.     

Ciprofloxacin in the treatment of pneumonia.

The use of ciprofloxacin as the sole agent in the treatment of 25 patients with pneumonias caused by susceptible organisms resulted in rapid cure. No side effects, superinfections, or recurrences were observed.     

Different modulation of decorin production by lung fibroblasts from patients with mild and severe emphysema

We have previously reported diminished immunohistochemical staining of decorin in lung tissue from patients with severe emphysema. The aim of this study is to investigate whether this diminished staining is due to a quantitative abnormal production of decorin by pulmonary fibroblasts in vitro. Therefore, we measured decorin (Western blot), collagen type I (ELISA), and fibronectin (ELISA) production by fibroblasts obtained from lung tissue of patients with severe and mild emphysema at basal culture conditions and after modulation with transforming growth factor-beta1, basic fibroblast growth factor, and interferon-gamma. Decorin production at basal culture conditions was significantly higher in fibroblast cultures from patients with severe emphysema compared to fibroblasts from mild emphysema. After stimulation with transforming growth factor-beta1 and basic fibroblast growth factor, decorin production was significantly more reduced in fibroblast cultures from patients with severe emphysema whereas collagen type I and fibronectin production were not affected. We conclude that decorin production by lung fibroblasts of patients with severe emphysema is dysregulated after modulation with cytokines known to be important in smoking associated inflammation. This dysregulation of decorin production may contribute to the impaired lung tissue repair, present in patients with emphysema, since these alterations in the extracellular matrix may cause diminished cytokine binding and neutralization.     

Killing of oxacillin-exposed staphylococci in human polymorphonuclear leukocytes.

Twelve strains of Staphylococcus aureus isolated from patients and two collection strains were grown on membranes placed on agar containing subminimal inhibitory concentrations of oxacillin. Clusters of staphylococci held together by thick cross walls resulted. These organisms, as well as the same strains grown in the same way on drug-free medium (control), were eluted from the membranes and were incubated with human polymorphonuclear leukocytes (PMNs) from various donors. Phagocytosis was comparable for both staphylococci exposed to oxacillin and control staphylococci, but the killing effect was different. The staphylococci grown on membranes in the presence of oxacillin were less susceptible to killing than the control staphylococci, but the killing effect was different. The staphylococci grown on membranes in the presence of oxacillin were less susceptible to killing than the control staphylococci. After 0.5 and 1 h of incubation with PMNs, the killing rates for oxacillin-grown versus control staphylococci were 52 and 70% and 65 and 85%, respectively (P < 0.01). After 2 and 3 h of incubation, the killing was similar. Most clusters of staphylococci contain a few individual cells that are located in the center of the cluster and are surrounded by other staphylococci; therefore, they are protected from adverse agents in the environment. This could explain why the phagocytized clusters are less susceptible than control staphylococci to the killing effect of PMNs during the first hour of incubation. Oxacillin does not penetrate into PMNs and in the absence of the drug the cross walls lyse, liberating the constituent staphylococci. This coincides with the increase in the percentage of cluster killing by PMNs after 2 and 3 h of incubation.     

Effect of serum on gram-positive cocci grown in the presence of penicillin.

Strains of Staphylococcus aureus, Streptococcus faecalis, Streptococcus bovis, and a strain of Streptococcus that produces filaments were grown on agar containing penicillin at concentrations of one-half to 1/10th the minimal inhibitory concentration. These penicillin-exposed organisms as well as untreated control organisms were incubated with human serum of plasma. Both serum and plasma produced a remarkable bactericidal effect on the filament-forming Streptococcus grown in the presence of penicillin, whereas the untreated control was only slightly affected. This response resembled that of gram-negative bacilli rather than that of gram-positive cocci. The growth of staphylococci exposed to penicillin was slightly inhibited in the presence of serum, whereas the growth of untreated staphylococci was stimulated. The streptococci, regardless of whether they were treated with penicillin or whether they were untreated, showed no change in growth pattern in the presence of serum.