Dietary zerumbone from shampoo ginger: new insights into its antioxidant and anticancer activity.

The dietary sesquiterpene dienone zerumbone (ZER) selectively targets cancer cells, inducing mitochondrial dysfunction and apoptosis, and protects non-cancerous cells towards oxidative stress and insult. This study examines the in vitro effects of ZER on lipid peroxidation in biological systems (cholesterol and phospholipid membrane oxidation) and explores its antitumor action in terms of its ability to modulate cancer cell lipid profile. Evaluation of the antioxidant activity of ZER showed that this compound is unable to trap lipoperoxyl radicals per se. ZER significantly modulated the total lipid and fatty acid profiles in cancer cells, inducing marked changes in the phospholipid/cholesterol ratio, with significant decreases in the levels of oleic and palmitic acids and a marked increase of stearic acid. Cell-based fluorescent measurements of intracellular membranes and lipid droplets using the Nile Red staining technique showed that in cancer cells, ZER induced significant accumulation of cytosolic lipid droplets and altered cell membrane organization/protein dynamics, depolarizing the mitochondrial membranes and inducing apoptosis and alteration of nuclear morphology. The modulatory activity of ZER on the total lipid and fatty acid profiles and lipid droplets may therefore represent another possible mechanism of its anticancer properties.

Mugil cephalus roe oil obtained by supercritical fluid extraction affects the lipid profile and viability in cancer HeLa and B16F10 cells.

We explored the changes in viability and lipid profile occurring in cancer cells, murine melanoma cells (B16F10 cells) and human cervical carcinoma cells (HeLa cells), when exposed to 24 h-treatments with an n-3 PUFA-rich oil obtained by supercritical extraction with CO2 from Mugil cephalus processed roe (bottarga). The composition of the major lipid classes of bottarga oil was determined by the (13)C NMR technique. Reversed-phase HPLC with DAD/ELSD detection was performed to analyze cells' total fatty acid profile and the levels of phospholipids, total/free cholesterol, triacylglycerols, and cholesteryl esters. Cell-based fluorescent measurements of intracellular membranes and lipid droplets were performed on bottarga oil-treated cells using the Nile red staining technique. The treatments of cancer cells with bottarga oil reduced the viability and affected the fatty acid profile, with a significant n-3 PUFA increase in treated cells. Mullet roe oil uptake modulated the cancer cell lipid composition, inducing a remarkable incorporation of health beneficial n-3 PUFA in the polar and neutral lipid fractions. Bottarga oil treatment influenced the synthesis of intracellular membranes and accumulation of cytoplasmic lipid droplets in cancer cells.

Localization of MTT formazan in lipid droplets. An alternative hypothesis about the nature of formazan granules and aggregates.

MTT (3-(4, 5-dimethyl-2-thiazolyl)-2, 5-dihphenyltetrazolium bromide) assay is a widely used method to assess cell viability and proliferation. MTT is readily taken up by cells and enzymatically reduced to formazan, a dark compound which accumulates in cytoplasmic granules. Formazan is later eliminated by the cell by a mechanisms often indicated as exocytosis, that produces characteristic needle-like aggregates on the cell surface. The shape of formazan aggregates and the rate of exocytosis change in the presence of bioactive amyloid beta peptides (Abeta) and cholesterol. Though the cellular mechanisms involved in MTT reduction have been extensively investigated, the exact nature of formazan granules and the process of exocytosis are still obscure. Using Nile Red, which stains differentially neutral and polar lipids, and a fluorescent analog of cholesterol (NBD-cholesterol), we found that formazan localized in lipid droplets, consistent with the lipophilic nature of formazan. However, formazan granules and aggregates were also found to form after killing cells with paraformaldehyde fixation. Moreover, formazan aggregates were also obtained in cell-free media, using ascorbic acid to reduce MTT. The density and shape of formazan aggregates obtained in cell-free media was sensitive to cholesterol and Abeta. In cells, electron microscopy failed to detect the presence of secretory vesicles, but revealed unusual fibers of 50 nm of diameter extending throughout the cytoplasm. Taken together, these findings suggest that formazan efflux is driven by physico-chemical interactions at molecular level without involving higher cytological mechanisms.

Preoperative chemoradiotherapy for oesophageal cancer: a systematic review and meta-analysis.

BACKGROUND: The benefit of neoadjuvant chemoradiotherapy in oesophageal cancer has been extensively studied but data on survival are still equivocal. OBJECTIVE: To assess the effectiveness of chemoradiotherapy followed by surgery in the reduction of mortality in patients with resectable oesophageal cancer. METHODS: Computerised bibliographic searches of MEDLINE and CANCERLIT (1970-2002) were supplemented with hand searches of reference lists. STUDY SELECTION: Studies were included if they were randomised controlled trials (RCTs) comparing preoperative chemoradiotherapy plus surgery with surgery alone, and if they included patients with resectable histologically proven oesophageal cancer without metastatic disease. Six eligible RCTs were identified and included in the meta-analysis. DATA EXTRACTION: Data on study populations, interventions, and outcomes were extracted from each RCT according to the intention to treat method by three independent observers and combined using the DerSimonian and Laird method. RESULTS: Chemoradiotherapy plus surgery compared with surgery alone significantly reduced the three year mortality rate (odds ratio (OR) 0.53 (95% confidence interval (CI) 0.31-0.93); p = 0.03) (number needed to treat = 10). Pathological examination showed that preoperative chemoradiotherapy downstaged the tumour (that is, less advanced stage at pathological examination at the time of surgery) compared with surgery alone (OR 0.43 (95% CI 0.26-0.72); p = 0.001). The risk for postoperative mortality was higher in the chemoradiotherapy plus surgery group (OR 2.10 (95% CI 1.18-3.73); p = 0.01). CONCLUSIONS: In patients with resectable oesophageal cancer, chemoradiotherapy plus surgery significantly reduces three year mortality compared with surgery alone. However, postoperative mortality was significantly increased by neoadjuvant chemoradiotherapy. Further large scale multicentre RCTs may prove useful to substantiate the benefit on overall survival.

Intra- and intercellular distribution of mitochondrial probes and changes after treatment with MDR modulators.

Fluorescent probes are currently used to evaluate the mitochondrial transmembrane potential in situ. However, in parallel experiments using the probes JC-1 and TMRM in different cell types (human astrocytes, HEp-2, Vero, KB, and HeLa cells), we found that the distribution of JC-1 and TMRM is highly variable not only in different cell types but also in different cells of the same cell type, a condition that has never been documented until our work. This phenomenon depends on a hidden, widespread multidrug resistance (MDR) phenotype that can be recognized only by comparative assays with MDR inhibitors (progesterone, verapamil, and cyclosporin A) and represents a serious risk of error in the evaluation of the mitochondrial potential.

Homogeneous longitudinal profiles and synchronous fluctuations of mitochondrial transmembrane potential.

This study reports for the first time (a) the longitudinal profile of the transmembrane potential (mDeltapsi) of single mitochondria using a Nernstian fluorescent probe and (b) the distribution of mDeltapsi fluctuations of mitochondria undergoing permanent depolarization. Our findings show that (1) mitochondria in different energetic conditions coexist in the same cell, (2) mDeltapsi is rather homogeneous along the entire length of single mitochondria, (3) mDeltapsi is not influenced by the surrounding cytoplasmic environment and (4) mDeltapsi fluctuations occur simultaneously in groups of mitochondria connected in a network. Taken together, these findings provide further evidence for a functional relationship between mitochondrial arrangement and energetic condition.

Diosmectite (DIOSMECTAL) in preventive anti-diarrheic therapy in patients subjected to pelvic radiation.

BACKGROUND: The radiation therapy of abdominal-pelvic tumors, determining diarrhea and cramps, could reduce the compliance of the patient and compromise his quality of life. This study attempts to assess the effects of adjuvant therapy with diosmectite (Diosmectal) on the above mentioned side effects from radiotherapy. METHODS: From August 1998 to September 1999, 40 patients (mean age 68) at the Oncological Radiotherapy Division of the Azienda Policlinico of Modena took part in the study: 21 forming the group of patients treated with diosmectite and 19 forming the control group. The radiotherapy was administered with total doses varying from 45 to 70 Gy in both groups with daily doses of 1.8-2 Gy. From the beginning of the therapy the patients in the group treated with Diosmectal, received 6 g daily doses of the drug split into two administrations. The diarrhea was assessed by referring to the Lent Soma score. RESULTS: In the group of patients not in therapy, an incidence of diarrhea was observed in 77% of subjects while in the group in therapy the incidence of diarrhea dropped to 25% of the cases (p<0.005), with an increase in the latency of the appearance of any diarrhoeic symptoms. CONCLUSIONS: Our data suggests that the administration of diosmectite in a prophylactic manner is capable of reducing the incidence of gastroenteric symptomatology (diarrhea and cramps).

Blepharophimosis, ptosis, epicanthus inversus syndrome, a new case associated with de novo balanced autosomal translocation [46,XY,t(3;7)(q23;q32)].

This paper reports a further case of blepharophimosis, ptosis, epicanthus inversus (BPES) syndrome associated with a reciprocal translocation [46,XY,t(3;7)(q23;q32)], involving band 3q23. This case supports the assignment of a BPES gene(s) to the 3q23 region.

Hypothalamic growth hormone deficiency in a patient with ring chromosome 18.

We report on a boy with a ring 18 chromosome associated with hypothalamic growth hormone (GH) deficiency. A 12-month trial of GH replacement therapy (0.5 U/kg/week) resulted in a marked growth acceleration. Our findings emphasise the need of evaluating GH secretion in patients with abnormalities of the 18 chromosome.

Prognostic significance of nucleolar organizer regions in ovarian epithelial tumors.

Recent studies have shown that the silver-stained nucleolar organizer region (AgNOR) score is related to the cell growth rate in several neoplasms. In the work presented here, we tested the AgNOR technique in 79 ovarian epithelial tumors (13 benign, 10 borderline, 56 malignant) to evaluate the diagnostic potential of AgNOR count in distinguishing between ovarian borderline tumors and carcinomas and to assess its prognostic value in carcinomas. Ovarian carcinomas exhibited higher mean AgNOR values than borderline and benign tumors, but statistically significant differences were found only in the serous type. Statistical analysis showed a positive correlation of higher AgNOR score, advanced tumor stage, and adverse prognosis. On the contrary, low AgNOR counts identified stages I and II carcinomas with disease-free follow-up. These results suggest that the AgNOR count may improve the prognostic evaluation of ovarian epithelial tumors by representing a reliable indicator of survival.

[Unknown primary bone metastasis: therapeutic and diagnostic strategies]. / Metastasi ossee con partenza sconosciuta: strategie terapeutiche e diagnostiche.

The primary site remains unknown in 0.5-15% of patients with multiple metastases. In 5-20% of these patients bone metastases are the first sign of disease. Survival after diagnosis ranges 2 to 6 months and 1-year survival is less than 25%. First of all, this paper reviews the current diagnostic and therapeutic approaches to patients with bone metastases from an unknown primary site and then reports the 10-year experience with radiotherapy at the Radiotherapy Division of the Centro di Riferimento Oncologico (Aviano) and of the Pordenone General Hospital in this patient population. Seventy-nine patients with bone metastases were evaluated. In 33 patients histology confirmed the original diagnosis and in 46 patients diagnosis was based on radiological and clinical findings. With respect to pain relief, differences in irradiated areas and in the ages of the patients were not significant. The patient's sex, however, did make a difference both to the degree of pain relief achieved and to survival (more favorable in women). To conclude, this paper suggests the diagnostic procedures necessary for the correct follow-up of patients with bone metastases from an unknown primary site.

Adjuvant radiotherapy of the pelvis with or without reduced glutathione: a randomized trial in patients operated on for endometrial cancer.

A randomized pilot trial was performed to evaluate the feasibility of administration of glutathione (GSH, 1200 mg, i.v.) as a protector in preventing diarrhea in patients operated on for endometrial cancer and submitted to adjuvant radiotherapy of the pelvis. Diarrhea occurred in 52% of patients in the untreated control group and only in 28% of patients in the GSH-treated group. Our preliminary data indicate that GSH administered before radiotherapy reduced the occurrence of diarrhea from oxidative damage to the intestinal mucosa. A large-scale phase III study is required to obtain definitive conclusions on the protective potential of GSH.

X-linked mental retardation and characteristic physical features in two brothers with duplication Xp22-Xpter.

Two brothers are reported who share mental retardation, conjunctival teleangectasias (mainly equatorial) and characteristic flat face with small mouth and thin prolabia. At the neuropsychological examination, the older brother at 14 years showed a full scale IQ of 40 (WISC), with verbal IQ 45 and performance IQ 44. The younger brother at 7 years showed a full scale IQ of 58 (WPPSI), with verbal IQ 67 and performance IQ 55. Chromosome studies showed a duplication Xp22-Xpter in both brothers and in the inactivated X of their mother. The anomaly was not present in a 3rd healthy brother and in other healthy relatives. The mother has normal intelligence and did not present any of the physical features of her affected sons.

Clinical evaluation of CA 15.3 in the post-operative follow-up of breast cancer patients.

In 265 patients operated for breast carcinoma the monoclonal antibody serum test CA 15.3 was predictive of metastatic diffusion of the disease. Its level increased in cases of distant metastasis with no significant difference between multiple and single sites (p = 0.014). The concentration of the marker was higher in 21 (23.8%) patients without nodal involvement and in 19 (27.5%) with nodal involvement (p = 0.193). Our study suggests that CA 15.3 may be an aid in the follow-up of patients with metastatic diffusion of breast cancer.

Prognostic significance of CA 15.3 in metastatic breast cancer.

We have investigated the possible relation between serum levels of CA 15.3 and disease status in 110 patients after radical mastectomy for breast cancer, with metastatic diffusion. Its persistent elevation was usually related to a very poor prognosis. In patients who died within 18 months the marker was always elevated. In case of progression of the disease, the marker level appeared to be consistently correlated with the general clinical condition. In healthy patients with stable disease the marker remained near the normal range.

Pseudomosaic centric fission of chromosome 4 in amniotic cells.

This paper describes a case of pseudomosaic centric fission of chromosome 4 detected in amniotic fluid cell culture. The pregnancy went to term and the newborn had a normal chromosomal constitution.