In view of the controversial information on the significance of the cyclin-dependent kinase inhibitor p21Cip1 in ovarian cancer, we conducted a retrospective investigation to clarify the relationships of this protein to proliferation rate, clinicopathological variables and prognosis of epithelial ovarian tumors.Paraffin-embedded tissue from 43 ovarian tumors of low malignant potential (LMP) and 82 primary ovarian adenocarcinomas were stained immunohistochemically for p21Cip1, p53 protein and Ki-67 antigen (a marker of cell proliferation).p21Cip1 levels were significantly higher in LMP tumors (p<0.001) as well as in early stage adenocarcinomas (p=0.021) and those associated with minimal residual disease (p=0.008). However, no relationship existed between p21Cip1 expression and the proliferation rate of adenocarcinomas or LMP tumors. In the vast majority of LMP tumors p21Cip1 expression was not accompanied by p53 accumulation. This p21Cip1-positive/p53-negative phenotype prevailed in the early stage (p=0.026), lower grade (p=0.018) adenocarcinomas as well as in those left with minimal residual disease (p=0.059). In patients with lower grade adenocarcinomas, decreased p21Cip1 expression was adversely related to poor overall survival on its own (p=0.0500) and when combined with p53 protein overexpression (p=0.0323). In multivariate analysis, only the stage remained as the independent predictor of survival.Decreased p21Cip1 expression is related to several indicators of aggressiveness in ovarian adenocarcinomas and seems to be differentially regulated in LMP tumors and adenocarcinomas. On the contrary, deregulation of p21Cip1 expression does not seem to participate in the pathogenesis of LMP tumors. Furthermore, although p21Cip1 alone or combined with p53 is of prognostic significance in lower grade adenocarcinomas, it does not appear to add to the information gained from traditional prognosticators.
Roberts syndrome (RS) is a rare autosomal recessive disorder characterized primarily by symmetric reduction anomalies of all limbs, growth retardation and craniofacial abnormalities. Most RS patients are reported to present a typical abnormality of their constitutive heterochromatin, accompanied by abnormal cytological growth characteristics. We present an extremely severe case of an RS fetus, karyotypically documented, with a clinical presentation including growth deficiency, tetraphocomelia, frontal meningocele, craniofacial abnormalities and penile enlargement with hypospadias. Nuclear morphometrical analysis in tissues of various organs revealed a reduced nuclear size in RS as compared to normal controls, and statistically significant differences in morphometric parameters related to the nuclear shape. Immunohistochemical study of the same organs showed a reduced expression of proliferating cell nuclear antigen in the presented case, thus indicating a decreased cell proliferation rate in RS. Our results reconfirm previously reported findings in cultured fibroblasts of RS cases, thereby reinforcing on a histologic level, the hypothesis that reduced cell proliferation may be involved in the growth retardation and the reduction abnormalities observed in RS.
In the rat, experimental manipulations that cause activation of the magnocellular neurosecretory neurones result in the synthesis, in addition to vasopressin ( AVP ) and oxytocin ( OXY ), of other neurotransmitters or peptides, including tyrosine hydroxylase ( TH ), the first and rate limiting enzyme for catecholamine biosynthesis. In the human neonate, our previous study showed that TH was selectively increased in AVP neurones of subjects that died from prolonged perinatal hypoxia. The purpose of the present study was to quantitatively investigate the expression of TH , AVP , OXY and neurophysin in magnocellular neurones of the human neonate in relation to the severity/duration of perinatal hypoxia, as estimated by neuropathological criteria. Autopsy was performed after obtaining parental written consent for diagnostic and research purposes. The intensity of the immunohistochemical reactions and the cellular/nuclear size were measured in the dorsolateral supraoptic nucleus using a computerised image analysis system. We showed that prolonged perinatal hypoxia resulted in the activation of the magnocellular neuroendocrine neurones of the human neonate, as indicated by their increased neuronal and nuclear size. OXY neurones appeared larger than the AVP ones at birth, possibly indicating an active role of foetal OXY during labour or even earlier. The gradual increase in the duration of the insult resulted in the reduction of intracellular AVP content, in parallel with a dramatic increase in the expression of TH , indicating a functional interaction of these peptides under neuronal activation. Ιsolated evidence in our series, obtained from an infant of a diabetic mother, raises the probability that in the case of hyperglycaemia the above pathogenetic mechanisms are diversified.
Giant ) is a rare placental tumor associated with complications including polyhydramnios, fetal anemia, cardiomegaly, hydrops and increased perinatal mortality1. Prenatal therapy may be performed when there are ultrasound features of fetal compromise and the gestation is not expected to survive. Therapeutic interventions include direct injection of various chemicals and laser coagulation of the tumor's feeding vessels2, 3. We report a case of a large chorioangioma located close to the placental cord insertion site, treated using interstitial laser coagulation. A 30-year-old patient presented at 24 weeks of gestation with polyhydramnios. On ultrasound, there was a large chorioangioma of 14 cm at its maximum diameter, arising from an anterior placenta within 1 cm of the placental cord insertion site (Figure 1). Three-dimensional Doppler ultrasound identified three deep feeding vessels and an additional large one running along the surface of the tumor connected directly to the umbilical cord confluence (Figure 2a). There was normal fetal growth, polyhydramnios with a deepest pool of 17 cm and mild fetal cardiomegaly. Peak systolic velocity (PSV) in the middle cerebral artery (MCA) was 51 cm/s (> 1.5 SD above the mean), indicating fetal anemia. Following counseling, the couple opted for vascular occlusion of the chorioangioma using interstitial laser photocoagulation. The procedure was performed under local anesthesia and continuous sonographic guidance. Routine prophylaxis with antibiotics (cephalexin, 2 g intravenously) was administered. A 17-gauge chorionic villus sampling needle (Chiba, Cook Inc., Bloomington, IN, USA) was inserted anteriorly, with the port of entry chosen to optimize visualization and operative access. A 1.07-mm non-contact laser fiber (Dornier MedTech GmbH, Wessling, Germany) was passed through the operative channel and coagulation was applied selectively to three feeding vessels, with 30–40-W intermittent beams depending on the diameter of the targeted vessel (Figure S1). The procedure was followed by amniodrainage of 3.5 L. One hour after the procedure, MCA-PSV was normal and the patient was discharged the next day. Five days later, there was significant sonographic evidence of degeneration within the tumor (Figure 2b). At 31 weeks of gestation, there was an acute drop in amniotic fluid followed by deterioration in fetal Dopplers. Cesarean section was performed for fetal distress after steroid administration to the mother. The neonate weighed 1760 g and was discharged healthy on day 40. Injection of toxic substances for the treatment of chorioangioma carries the risk of potential fetal exposure, to which at least one neonatal death has been attributed2. Laser photocoagulation can be performed either fetoscopically3 or using an ultrasound-guided interstitial approach4. Fetoscopy has a reported success rate of 60–80%; however, this technique may be difficult to perform in cases of anterior placenta3, 5. An interstitial approach is less invasive, using a device with a smaller diameter, and, in three of a series of cases in which it was used, was successful in all cases4. In the series of Zanardini4, the treated chorioangiomas were between 3.5 and 5.4 cm in diameter and close proximity to the cord was considered a contraindication for an interstitial approach in some cases because of a fear of possible excess thermal damage. The chorioangioma in the present case is, to the best of our knowledge, the largest reported one to be treated and the first treated by interstitial laser coagulation despite being located adjacent to the placental cord insertion site (Figure S2). This approach may be preferable in anterior placentas. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
Disruption of apoptotic cell death has been implicated in tumour aggressiveness in colonic carcinogenesis. The Fas-Fas ligand (FasL) system is involved in the execution of apoptosis induced by the immune system. c-FLIP protein constitutes an inhibitor of Fas and other (TRAIL) death receptor-mediated apoptosis. The aim of this study was to investigate the simultaneous expression of Fas, FasL and c-FLIP in relation to standard clinicopathological parameters and patients' outcome in colorectal cancer.Levels of Fas, FasL and c-FLIP protein expression were quantified immunohistochemically in paraffin-embedded tissues from 90 patients. Immunopositivity was detected for Fas, FasL and c-FLIP in 71%, 35.5% and 68.8% of cases, respectively. Concurrent expression of Fas/FasL was seen in 28 samples (31%), of which 24 (85.7%) also displayed c-FLIP positivity (P = 0.04). c-FLIP overexpression (> 10%) tended to prevail marginally in higher stage tumours (P = 0.09). Additionally, FasL and c-FLIP adversely affected survival on both univariate (P = 0.001 and P = 0.0024, respectively) and multivariate analysis [hazard ratio (HR) 3.491, P = 0.005 and HR 2.960, P = 0.036, respectively].The frequent expression and coexpression of Fas, FasL and c-FLIP in colorectal carcinoma implicates c-FLIP as an inhibitor of the Fas-FasL-induced death pathway in these tumours. Moreover, c-FLIP conveys independent prognostic information in the presence of classical prognosticators.
Hypoxia‐inducible factor (HIF)‐1α is a transcription factor that promotes ischaemia‐driven angiogenesis. The aim of this study was to determine the relation of HIF‐1α to vascular endothelial growth factor (VEGF; an important angiogenic molecule in brain tumours), p53 expression, angiogenesis, proliferative potential and clinical outcome in a large series of diffuse astrocytomas. Expression of HIF‐1α, VEGF, Ki‐67 (a proliferation‐associated marker) and p53 was determined immunohistochemically in 83 adult patients with supratentorial diffuse astrocytomas. Microvessels, highlighted by means of anti‐CD34 immunohistochemistry, were enumerated with computer‐assisted image analysis. Although HIF‐1α and VEGF were expressed in the majority of cases, their levels increased significantly with increasing grade and proliferative potential. HIF‐1α positively correlated with microvessel counts and VEGF with total vascular area and the presence of rounder vessel sections. There was a positive correlation of VEGF with p53 expression in astrocytomas and anaplastic astrocytomas. In univariate analysis, both VEGF and HIF‐1α were associated with shortened survival in the entire cohort, but lost significance when grades II/III and grade IV were analysed separately. Multivariate analysis revealed that the combination of HIF‐1α with grade was a significant prognostic indicator. HIF‐1α expression may be used to refine the prognostic information provided by grade in patients with diffuse astrocytomas. Its adverse prognostic effect is most likely mediated by hypoxia, the driving force for HIF‐1α accumulation.
Objective To examine the effects of short-term cyclic stretch on apoptosis in alveolar type II cells (A549).To study in vitro the direct influence of alveolar type II cells on mechanical stretch.Methods A549 were treated with different doses of lipopolysaccharide (LPS), 0 ng/ml, 1 ng/ml, 10 ng/ml, 100 ng/ml, 1000 ng/ml, and then A549 were lengthened 5%, 15%, 30% using a FLEXCELL tension unit 4000, a vacuum-driven device that applies strain to cells, which were cultured in six-well plates coated with collagen-I, and 12 cycles/min for 4 hours.Apoptosis was measured using the flow cytometry method that measures annexin V and propidium iodide (PI) staining.The morphological changes of apoptotic cells were observed by transmission electron microscope.Results Apoptosis could be induced in alveolar type II cells (A549) by mechanical stretch.The percentage of annexin V + PI cells increased after being treated with cyclic stretch for 4 hours by 5%, 15%, 30% in all groups.The morphological features of apoptotic cells demonstrated by transmission electron microscope were as follows: shrinkage of the cell, chromatin condensation and aggregation under the nuclear membrane as a crescent or lump, membrane-encapsulated nuclear fragment or cell organ formed by invagination of the cell membrane, and apoptotic body formation followed by vacuolization.Conclusion Apoptosis induced by mechanical stretch and LPS is dose dependent.Mechanical stretch aggravates apoptosis especially in cells treated with LPS.Annexin V and PI double staining is a specific, sensitive, and quantitative method for analyzing apoptotic cells.It is also helpful to clarify the protective mechanism of low-volume ventilation in ARDS.
