High occurrence of Anisakidae at retail level in cod (Gadus morhua) belly flaps and the impact of extensive candling.

The presence of Anisakidae at retail level, after the routine screening via candling, was investigated in cod, the most commonly consumed fish species in Belgium. A total of 780 pre-packed belly flap samples destined for one branch of retail shops were collected from a Belgian wholesale company. To recover all larvae, each sample was first candled and thereafter enzymatically digested. Larvae were morphologically identified to the genus level and a subset was additionally molecularly confirmed by amplification of the ITS fragment and HinfI/HhaI enzyme restriction. The PCR/RFLP profiles of Contracaecum spp. were determined and confirmed with sequencing by the European Reference Laboratory for Parasites (Istituto Superiore di Sanità). The positivity rate of Anisakidae in the individual cod samples was 18% [95%-CI: 15-21%], with a mean intensity of one larva [range: 1-6]. Belly flaps were sold packed primarily by two, with a one-in-three chance of buying an infected package. Pseudoterranova spp. infections (single infections) were most frequently detected (positivity rate 9% [95%-CI: 7-11]), closely followed by Anisakis spp. (7% [95%-CI: 6-9]). Co-infections of Pseudoterranova spp. and Anisakis spp. comprised 8% of the infections, with a positivity rate of 1% [95%-CI: 1-3%]. All belly flaps reportedly were candled prior to our sampling, nonetheless our results indicated that an additional candling screening before packaging would identify an extra third of the infections and larvae. In 19 of the 139 infected samples, all larvae were recovered by the additional candling, thereby removing the infection risk for consumers. In conclusion, this study shows that cod belly flaps infected with zoonotic parasites reach the Belgian consumer. Although a second candling step at retail level could be helpful in reducing the consumer risk, additional measures are needed since 66% of infections would still remain undetected.

Preliminary results of phase II study of capecitabine and gemcitabine (CAP-GEM) in patients with metastatic pretreated thymic epithelial tumors (TETs).

BACKGROUND: No previous prospective trials have been reported with capecitabine and gemcitabine (CAP-GEM) in patients with metastatic thymic epithelial tumors (TETs). We conducted a multicenter study to determine the activity and tolerability of this regimen in pretreated TETs. PATIENTS AND METHODS: A total of 15 patients were enrolled in the first stage of phase II study. All patients received CAP-GEM every 3 weeks. The primary end point was objective response rate (RR); secondary end points were toxicity, progression-free survival (PFS) and overall survival. RESULTS: Complete responses (CR) and partial responses were observed in three (20%) and three (20%) patients for a 40% RR, respectively. Grade 1-2 neutropenia, anemia and thrombocytopenia were the most common side-effects, noted in seven (46.7%), five (33.3%) and five (33.3%) patients, respectively. The most common grade 3 toxicity was neutropenia in three patients (20%). Median PFS was 11 months (95% confidence interval 4-17). The 1- and 2-year survival rates were 80% and 67%, respectively. CONCLUSION: We have decided to publish the preliminary results because this regimen was more active than that expected. Although our results are preliminary, CAP-GEM shows activity and safety in pretreated TETs. Furthermore, multicenter trials, also in first-line setting, are necessary to confirm our results.

Multilocus genotyping of Giardia duodenalis reveals striking differences between assemblages A and B.

Giardia duodenalis is a widespread parasite of mammalian species, including humans. Due to its invariant morphology, investigations of aspects such as host specificity and transmission patterns require the direct genetic characterisation of parasites from faecal samples. We performed a sequence analysis of four genes (ssrRNA, ß-giardin, glutamate dehydrogenase and triose phosphate isomerase) of 61 human isolates and 29 animal isolates. The results showed that multilocus genotypes (MLGs) can be readily defined for G. duodenalis isolates of assemblage A but not for assemblage B. Indeed, for assemblage A isolates, there was no evidence of intra-isolate sequence heterogeneity, and congruent genotyping results were obtained at the four genetic loci investigated. Sequence comparison and phylogenetic analysis showed that human-derived and animal-derived MLGs are different, and further indicated the presence of a new sub-assemblage (referred to as "AIII"), which was found exclusively in wild hoofed animals. On the other hand, there were variable levels of intra-isolate sequence heterogeneity (i.e., the presence of two overlapping nucleotide peaks at specific positions in the chromatograms, or "heterogeneous templates") in assemblage B isolates from humans and animals, and this prevented the unambiguous identification of MLGs. Furthermore, in five human isolates and one non-human primate isolate, the assignment to assemblage B was problematic, given that one of the four markers supported an assignment to assemblage A. These findings raise concerns about the interpretation of genotyping data based on single markers, and indicate the need to understand the mechanisms that are responsible for the differences between G. duodenalis assemblages A and B.

[Medical treatment for thymoma: an update]. / La terapia medica del timoma: update.

Stage of disease at diagnosis and histologic type according to WHO classification are the most important prognostic factors for thymoma and complete surgical resection represents a crucial point for disease free survival. When surgery is not feasible, neoadjuvant or palliative chemotherapy, is the most appropriate treatment because of the high chemosensitivity of the thymoma. The role of predictive factors to response of treatment seems relevant: the presence of modifications of tumor related genes and expression of different thymoma cell receptors could allow to identify subsets of patients who can benefit from target therapies, with the aim of optimizing treatments and improving survival.

A novel Giardia duodenalis assemblage A subtype in fallow deer.

The molecular identification of species and genotypes of Giardia spp. infecting wild mammals represents the most reliable tool to understand the role played by these animals as reservoirs of cysts infectious for human and other animals. Of 139 fecal samples collected from fallow deer (Dama dama L.) hunted in a Natural Reserve of northern Italy, the prevalence of Giardia sp. was 11.5% (16 of 139 animals), and it was higher in fawns than in older animals. Fragments of the betagiardin and triose phosphate isomerase (tpi) genes were successfully polymerase chain reaction amplified and sequenced from 8 isolates. No sequence variation was observed between isolates at the 2 genetic loci. Sequence and phylogenetic analyses identified a Giardia duodenalis subtype that clusters with assemblage A isolates and that shows homologies of 98 and 97% at the beta-giardin and tpi loci, respectively, compared with the A1 subtype. Because the G. duodenalis subtype found in fecal samples of fallow deer has never been detected previously, its role as a pathogen for humans and domestic animals is unknown, but, considering its genetic distinctiveness, it is likely to be low.

Epoetin alfa 40000 U once weekly and intravenous iron supply in solid tumor patients: early increase of hemoglobin level during chemotherapy.

The objective of this observational study was the early evaluation of the impact, a week after the first administration of epoetin alfa 40000 U once weekly and i.v. dose of 62.5 mg sodium ferric gluconate for seven days in improving hemoglobin levels in cancer patients affected by mild/moderate or severe anemia during chemotherapy. Twenty patients affected by solid tumors who received epoetin alfa 40000 U once weekly and daily i.v. sodium ferric gluconate for one week were evaluated: 90% of the patients showed hemoglobin increase, with a median level of hemoglobin increase of 0.73 g/L from baseline, and 50% of them showing a hemoglobin increase > 1 gr/L. The treatment was well tolerated and no adverse event was observed. The early increase of hemoglobin level from baseline is interesting and suggestive for the possibility of achieving an adequate hemoglobin level with a short-term treatment. It is still necessary to further explore the real need of iron supplementation to maintain adequate erythropoiesis prior and during epoetin therapy.

Dientamoeba fragilis is more prevalent than Giardia duodenalis in children and adults attending a day care centre in Central Italy.

Giardia duodenalis is a well recognised enteropathogen, while Dientamoeba fragilis is rarely detected and consequently it is not recognised as an important human pathogen. In 2002-2003, a survey has been carried out on enteroparasites in faecal samples of outpatients attending a day care centre in the town of Perugia (Central Italy). To improve the detection level, at least three samples from each patient were collected at different days and within two hours from defecation. The coproparasitological examination has been carried out by direct microscopic examination, faecal concentration, and Giemsa and modified Ziehl-Nielsen stainings of faecal smears. The genotypes of Giardia duodenalis isolates were determined by PCR of the beta-giardin gene. Of 1,989 enrolled people (966 children, 1,023 adults), 165 persons (8.3%; 153 adults, 15.0%; 12 children, 1.2%), were positive for parasites, but only 1 12 adults (73.2% of those infected) and eight children (66.7% of those infected) harboured D. fragilis and G. duodenalis. Both the Assemblages A and B were detected in 18 G. duodenalis isolates examined at the beta-giardin gene. The higher prevalence of D. fragilis infections than that of G. duodenalis is probably related to the method used, a procedure, which is rarely followed in laboratories for the diagnosis of enteric parasites. These epidemiological data suggest that when faecal samples are examined after a period of time and without Giemsa staining, most D. fragilis infections goes undetected.

[High-dose chemotherapy for metastatic breast cancer]. / La chemioterapia ad alte dosi nel carcinoma della mammella metastatico.

In ninety's breast cancer was first in Europe for the use of high-dose chemotherapy with autologous hematopoietic stem cell transplantation in solid tumors in adults. Some phase II trials of high-dose chemotherapy showed high response rates and prolonged progression free survival in selected metastastic breast cancer patients. Few large, powerful randomized phase III studies comparing this approach with conventional chemotherapy have been completed: some studies showed a better progression free survival in favor of high dose chemotherapy, but no statistically significant difference in overall survival was observed. Many variables inside high dose chemotherapy program need to be considered. The identification of subsets of breast cancer patients who can benefit from high-dose chemotherapy is essential: high-dose chemotherapy should be included in a global treatment strategy, evaluating the integration with innovative treatment modalities, with the aim of eradicating the minimal residual disease in breast cancer patients achieving complete response after high dose chemotherapy.

Anesthetic management of liver transplantation in patients with fulminant hepatic failure: case reports.

Fulminant hepatic failure (FHF), although not frequent, produces a high mortality rate of 70% to 90%. This study describes the management of FHF patients without the use of any intracranial pressure monitoring device.

Autologous bone marrow transplantation with negative immunomagnetic purging for aggressive B-cell non-Hodgkin's lymphoma in first complete remission.

To evaluate the effect on survival of negative immunomagnetic purging in aggressive B-cell non-Hodgkin's lymphoma (NHL), 20 patients retrospectively staged according to the age-adjusted International Prognostic Index as high-intermediate (11 patients) or high-risk (9 patients) received autologous bone marrow transplantation (ABMT) in first complete remission (CR1). All patients received six to eight cycles of a F-MACHOP-like protocol as induction treatment and then underwent high-dose chemotherapy (HDC) with a CBV-like regimen. Negative purging included a panel of monoclonal antibodies against B-cell antigens and immunomagnetic beads. The data were compared to a historical control of 18 patients with the same characteristics treated in our institution who received unpurged bone marrow support. The median yield of mononuclear cells (MNC), colony-forming units-granulocyte/macrophage (CFU-GM), and CD34+ cells after purging were 52%, 49%, and 57%, respectively. The median B-cell depletion after negative selection was 1.8 logs. All patients obtained a complete engraftment with no significant differences between the purged and unpurged group. Two toxic deaths (one for each group) were observed and the main extrahematological toxicities were mucositis, vomiting, and diarrhea. The event-free survival (EFS) and overall survival (OS) at 3 years for the whole group of 38 patients were 73% (95% CI: 59-88%) and 81% (95% CI, 68-94%), respectively. The comparison between patients receiving purged marrow and patients receiving unmanipulated marrow indicated no significant survival differences between the two groups both for EFS 84% (95% CI: 67-100%) vs 61% (95%CI: 39-84%) ( P=0.12) and OS 84% (95% CI: 69-100%) vs 71% (95% CI: 50-93%) ( P=0.58). Our report shows that HDC followed by reinfusion of autologous bone marrow can produce long EFS and OS in high-intermediate and high-risk patients with B-cell NHL transplanted in CR1, but was not be able to demonstrate a significant clinical advantage using immunomagnetic purged marrow. However, the use of ex vivo negative purging combined with innovative treatment modalities (peripheral blood stem cell transplant, in vivo administration of monoclonal antibodies) needs to be explored.

Administration of low-dose interleukin-2 plus G-CSF/EPO early after autologous PBSC transplantation: effects on immune recovery and NK activity in a prospective study in women with breast and ovarian cancer.

This study evaluated the effects of low-dose IL-2 plus G-CSF/EPO on post-PBSC transplantation (PBSCT) immune-hematopoietic reconstitution and NK activity in patients with breast (BrCa) and ovarian cancer (OvCa). To this end, two consecutive series of patients were prospectively assigned to distinct post-PBSCT cytokine regimens (from day +1 to day +12) which consisted of G-CSF (5 microg/kg/day) plus EPO (150 IU/kg/every other day) in 17 patients (13 BrCa and 4 OvCa) or G-CSF/EPO plus IL-2 (2 x 10(5) IU/m(2)/day) in 15 patients (10 BrCa and 5 OvCa). Hematopoietic recovery and post-transplantation clinical courses were comparable in G-CSF/EPO- and in G-CSF/EPO plus IL-2-treated patients, without significant side-effects attributable to IL-2 administration. In the early and late post-transplant period a significantly higher PMN count was observed in G-CSF/EPO plus IL-2-treated patients (P = 0.034 and P = 0.040 on day +20 and +100, respectively). No significant differences were found between the two groups of patients in the kinetics of most lymphocyte subsets except naive CD45RA(+) T cells which had a delayed recovery in G-CSF/EPO plus IL-2 patients (P = 0.021 on day +100). No significant difference was observed between NK activity in the two different groups, albeit a significantly higher NK count was observed in G-CSF/EPO plus IL-2 series on day +20 (P = 0.020). These results demonstrate that low-dose IL-2 can be safely administered in combination with G-CSF/EPO early after PBSCT and that it exerts favorable effects on post-PBSCT myeloid reconstitution, but not on immune recovery.

Costs of high-dose chemotherapy and peripheral blood progenitor cell autograft for breast cancer.

The aim of the study was to analyze the real cost of single or tandem high-dose chemotherapy (HDC) and peripheral blood progenitor cell autologous transplant (PBPCT) in patients with breast cancer. We analyzed the costs of 40 PBPCT performed in 20 patients. Tandem transplant was planned for each patient. Resources used and direct costs were identified for each patient. The study was carried out using the hospital perspective and monetary values were reported in 1999 Euro. The mean cost of whole procedure for single transplant was 20,816.63 Euro, while the mean cost of tandem transplant was 38,770.83 Euro. The cost distribution in the two groups was similar: the most expensive phase of procedure was the supportive phase post transplant (about 60% of total cost), with the categories of cost most represented being professional fees (about 28%) and pharmacy (about 35%). Awaiting more convincing trials of the clinical advantage of HDC in breast cancer, our analytical evaluation of transplant costs for different therapeutic options, single or tandem, permits identification of the most expensive categories in order to intervene for cost savings.

Docetaxel and epirubicin plus G-CSF as mobilizing treatment to support high-dose chemotherapy in breast cancer.

BACKGROUND: In order to combine an active regimen with a simultaneous efficient mobilization of peripheral blood precursor cells (PBPC), we explored the combination of Docetaxel 75 mg/m2 and Epirubicin 120 mg/m2 with G-CSF 5 mcg/Kg/day s.c. to mobilize PBPC in breast cancer patients to support high-dose chemotherapy (HDC). PATIENTS AND METHODS: Forty patients were enrolled: 27 high risk and 13 metastatic. The entire procedure, including chemotherapy and PBPC collection, was on an outpatient basis. RESULTS: The median day of starting apheresis was day +10 (range 10-12) and the average value of circulating CD34+ cells at peak was 175/microliter (range 33-403). The median yield of CD34+ cells per apheresis was 8.76 x 10(6)/Kg (range 1.83-27.87). None of the patients developed side effects which required hospitalization. All patients enrolled successively received HDC as consolidation treatment. High risk patients received one and metastatic patients two HDC with PBPC reinfusion. All patients obtained a complete engraftment. No significant differences between high-risk and metastatic patients were observed. CONCLUSIONS: Our study suggests that the combination of Docetaxel, Epirubicin, and G-CSF is feasible, safe and efficient outpatient mobilizing treatment for patients with breast cancer receiving HDC.

Autologous bone marrow transplantation in relapses of chemotherapy-sensitive aggressive non-Hodgkin's lymphoma: long-term outcome.

Eleven patients with relapsed intermediate to high grade non-Hodgkin's lymphoma (NHL) responding to induction treatment were treated with high-dose chemotherapy (CBV or ICBV conditioning regimen) plus autologous bone marrow transplantation as early consolidation treatment. At 6 years, relapse-free survival is 27.3% and overall survival is 36.4%. Patients with bone marrow involvement from NHL before the induction therapy did not have a worse prognosis. Despite the long-term follow-up, no secondary myelodysplasia or acute leukemia occurred in our patients. Within the limitations of patient number and selection, our retrospective study confirms the importance of tumor responsiveness and long-term follow-up. Patients with relapsed, but chemotherapy-sensitive NHL can achieve prolonged survival after high-dose chemotherapy plus autologous bone marrow transplantation.

BRCA1 and BRCA2 mutations in central and southern Italian patients.

Protein truncation test (PTT) and single-strand conformation polymorphism (SSCP) assay were used to scan the BRCA1 and BRCA2 genes in 136 unrelated Italian breast/ovarian cancer patients. In the sample tested, BRCA1 and BRCA2 equally contributed to site-specific breast cancer patients who reported one to two breast cancer-affected first-/ second-degree relative(s) or who were diagnosed before age 40 years in the absence of a family history of breast/ovarian cancer. BRCA1 and BRCA2 mutations were mostly found in patients with disease diagnosis before and after age 50 years, respectively. Moreover, in cases with familial clustering of site-specific breast cancer, BRCA1 mostly accounted for tumours diagnosed before age 40 years and BRCA2 for tumours diagnosed after age 50 years. The BRCA1 and BRCA2 mutation spectrum was consistent with a lack of significant founder effects in the sample of patients studied.

Autologous bone marrow transplantation as consolidation therapy in newly diagnosed non-Hodgkin's lymphoma: long-term outcome.

Autologous bone marrow transplantation (ABMT) often produces durable remission in patients with intermediate-high grade non-Hodgkin's lymphoma (NHL). We present a retrospective review of 32 eligible newly diagnosed patients with NHL treated with conventional induction chemotherapy followed by ABMT consolidation therapy. These patients were treated in our department between 1984-1994 and followed up for 5-172 months with a median time of 82 months. In our patients the status of disease at transplant was 30 complete remissions and 2 partial remissions. All patients received a CBV-like high-dose preparative regimen. At 136 months the probability of disease-free survival (DFS) and overall (OS) is 66% and 70% respectively. Seven patients died from the disease. There was one case of toxicity related death. Our aim was to achieve a status of minimal disease and then consolidate it with high-dose polychemotherapy regimen. This study confirms that a significant number of patients with aggressive responding NHL can achieve prolonged RFS and OS after ABMT. Our data document the importance of long-term follow-up in interpreting the results of ABMT in NHL.

Detection of breast cancer cells in the bone marrow or peripheral blood: methods and prognostic significance.

Tumor cells can reach every anatomic district, organ and tissue through the peripheral blood circulation. Tumor cell shedding is considered an early event in the multi-phase process of metastasis, and the possibility of detecting tumor cells in the bloodstream and/or bone marrow before clinical evidence of distant metastases needs to be explored. The use of new sophisticated diagnostic and investigative techniques has boosted the study of tumor cell contamination of bone marrow and peripheral blood. Molecular techniques, such as reverse-transcriptase polymerase chain reaction, may be useful tools to reach this target, but, today, immunocytochemistry is still considered the gold standard to assess new techniques to detect isolated tumor cells in hematopoietic tissue. Little is known about the biology of isolated tumor cells in the peripheral blood or bone marrow. Two crucial points need to be evaluated: the identification of specific markers of breast cancer cells with clonogenic potential and their biologic properties, and the prognostic impact of the detection of isolated tumor cells in the bone marrow or peripheral blood stem cell collections.

Pregnancy-associated breast cancer.

Pregnancy-associated breast cancer, which is defined as all breast cancer diagnosed during pregnancy or within the following year, is a relatively rare finding. Due to the particular difficulties in the diagnosis of breast cancer during this period, pregnant women tend to present more advanced disease at diagnosis. Four cases of pregnancy-associated breast cancer referred to our Institute are described as a contribution to the knowledge of this disease.

Alternative insertion sites for permanent central venous access devices.

Permanent central venous access devices (PCVAD) are used widely in the management of chronically ill patients, particularly in neoplastic diseases. The standard approach consists of positioning the catheter in the superior vena cava (SVC) either using subclavian or internal jugular vein puncture, or cephalic or external jugular vein cut-down, with the port implanted in a subcutaneous pouch of the thoracic region. Alternative insertion sites could be used in selected cases. In our experience, consisting of 158 PCVAD, 12 cases required a different insertion site: six cases of an SVC catheter and port on the forearm using a basilic vein cut-down, and six cases of an inferior vena cava (IVC) catheter and port in the abdominal region using a great saphenous vein cut-down. Comparing standard to alternative approaches, we observed a total morbidity rate of 8.9% and 8.3%, respectively (P=NS), while the explant rate was 5.4% vs 8.3% (P=0.1). Our data show non-significant differences in morbidity and explant rates between the two groups of patients. Alternative insertion sites for the PCVAD implant seem to be a valid possibility in the management of chronically ill patients.