Low-Avidity Autoantibodies against Bactericidal/Permeability-Increasing Protein Occur in Gram-Negative and Gram-Positive Bacteremia.

Antibody autoreactivity against bactericidal/permeability-increasing protein (BPI) is strongly associated with Pseudomonas aeruginosa infection in cystic fibrosis (CF), non-CF bronchiectasis (BE), and chronic obstructive pulmonary disease (COPD). We examined the pathogen-specific nature of this autoreactivity by examining antibodies to BPI in bacteremia patients. Antibodies to BPI and bacterial antigens were measured in sera by ELISA from five patient cohorts (n = 214). Antibody avidity was investigated. Bacteremic patient sera (n = 32) exhibited IgG antibody autoreactivity against BPI in 64.7% and 46.7% of patients with positive blood cultures for P. aeruginosa and Escherichia coli, respectively. Autoantibody titers correlated with IgG responses to bacterial extracts and lipopolysaccharide (LPS). A prospective cohort of bacteremic patient sera exhibited anti-BPI IgG responses in 23/154 (14.9%) patients with autoreactivity present at the time of positive blood cultures in patients with Gram-negative and Gram-positive bacteria, including 8/60 (13.3%) patients with Staphylococcus aureus Chronic tissue infection with S. aureus was associated with BPI antibody autoreactivity in 2/15 patients (13.3%). Previously, we demonstrated that BPI autoreactivity in CF patient sera exhibits high avidity. Here, a similar pattern was seen in BE patient sera. In contrast, sera from patients with bacteremia exhibited low avidity. These data indicate that low-avidity IgG responses to BPI can arise acutely in response to bacteremia and that this association is not limited to P. aeruginosa This is to be contrasted with chronic respiratory infection with P. aeruginosa, suggesting that either the chronicity or the site of infection selects for the generation of high-avidity responses, with biologic consequences for airway immunity.

Does the addition of oxaliplatin to preoperative chemoradiation benefit cT4 or fixed cT3 rectal cancer treatment? A subgroup analysis from a prospective study.

BACKGROUND: Whether there is any benefit derived from adding oxaliplatin to fluoropyrimidine-based preoperative chemoradiation is currently unknown in cases of advanced cT3 or cT4 tumours. Our aim was to evaluate this issue by analysing a randomized trial, which compared two schedules of preoperative treatment (chemoradiation vs. 5 × 5 Gy with 3 cycles of consolidation chemotherapy) for cT4 or fixed cT3 rectal cancer. PATIENTS AND METHODS: Delivery of oxaliplatin was mandatory to the first part of the study. For the second part, its delivery in both treatment-assigned groups was left to the discretion of the local investigator. We analysed a subgroup of 272 patients (136 in the oxaliplatin group and 136 in the fluorouracil-only group) from institutions that had omitted oxaliplatin in the second part of the study. RESULTS: Circumferential resection margin negative (CRM-) status rate was 68% in the oxaliplatin group and 70% in the fluorouracil-only group, p = 0.72. The pathological complete response rate (pCR) was correspondingly 14% vs. 7%, p = 0.10. Following multivariable analysis, when comparing the CRM- status in the oxaliplatin group to the fluorouracil-only group, the odds ratio was 0.79 (95 CI 0.35-1.74), p = 0.54; there being no interaction between concomitant chemoradiation and 5 × 5 Gy with consolidation chemotherapy; pinteraction = 0.073. For pCR, the corresponding results were 0.47 (95 CI 0.19-1.16), p = 0.10, pinteraction = 0.84. CONCLUSION: No benefit was found of adding oxaliplatin in terms of CRM nor pCR rates for either concomitant or sequential settings in preoperative radiochemotherapy for very advanced rectal cancer.

Long-course oxaliplatin-based preoperative chemoradiation versus 5 × 5 Gy and consolidation chemotherapy for cT4 or fixed cT3 rectal cancer: results of a randomized phase III study.

BACKGROUND: Improvements in local control are required when using preoperative chemoradiation for cT4 or advanced cT3 rectal cancer. There is therefore a need to explore more effective schedules. PATIENTS AND METHODS: Patients with fixed cT3 or cT4 cancer were randomized either to 5 × 5 Gy and three cycles of FOLFOX4 (group A) or to 50.4 Gy in 28 fractions combined with two 5-day cycles of bolus 5-Fu 325 mg/m(2)/day and leucovorin 20 mg/m(2)/day during the first and fifth week of irradiation along with five infusions of oxaliplatin 50 mg/m(2) once weekly (group B). The protocol was amended in 2012 to allow oxaliplatin to be then foregone in both groups. RESULTS: Of 541 entered patients, 515 were eligible for analysis; 261 in group A and 254 in group B. Preoperative treatment acute toxicity was lower in group A than group B, P = 0.006; any toxicity being, respectively, 75% versus 83%, grade III-IV 23% versus 21% and toxic deaths 1% versus 3%. R0 resection rates (primary end point) and pathological complete response rates in groups A and B were, respectively, 77% versus 71%, P = 0.07, and 16% versus 12%, P = 0.17. The median follow-up was 35 months. At 3 years, the rates of overall survival and disease-free survival in groups A and B were, respectively, 73% versus 65%, P = 0.046, and 53% versus 52%, P = 0.85, together with the cumulative incidence of local failure and distant metastases being, respectively, 22% versus 21%, P = 0.82, and 30% versus 27%, P = 0.26. Postoperative and late complications rates in group A and group B were, respectively, 29% versus 25%, P = 0.18, and 20% versus 22%, P = 0.54. CONCLUSIONS: No differences were observed in local efficacy between 5 × 5 Gy with consolidation chemotherapy and long-course chemoradiation. Nevertheless, an improved overall survival and lower acute toxicity favours the 5 × 5 Gy schedule with consolidation chemotherapy. CLINICAL TRIAL NUMBER: The trial is registered as ClinicalTrials.gov number NCT00833131.

YMn2Hx and RMn(2-y)Fe(y)H6 (R = Y, Er) studied by Raman, infrared and inelastic neutron scattering spectroscopies.

YMn2 forms either interstitial YMn2Hx hydrides for x < or = 4.5 or a complex YMn2H6 hydride when submitted to high hydrogen pressure. These compounds have been studied by inelastic neutron scattering (INS) in order to clarify the different modes of H vibration. The INS spectra of YMn2Hx hydrides are strongly dependent on the H content. YMn2H6 and YMn2D6 show broad bands, also observed by Raman and IR spectroscopy, assigned to H-Mn-H (or D) and Mn-H bending and stretching modes. Both ErMn2D6 and ErMn1.8Fe0.2D6 show, in addition to the H vibration mode, an intense band at 215 cm(-1) which has been attributed to a magnetic excitation of Er3+ in view of its momentum transfer dependence.

Formation of hydrides in (Ti(1-x)Zr(x))Co(2.00) (0 < x < 1) pseudobinary alloys.

The exposure of (Ti(1-x)Zr(x))Co(2.00) intermetallic alloys to hydrogen at high pressure caused (Ti(1-x)Zr(x))Co(2.00) (x = 0.50-0.90) hydrides in the alloy. The crystalline structural, electronic, and magnetic properties of parent alloys and of their hydrides were determined by using XRD (X-ray powder diffraction) and XAS (X-ray absorption spectrometry) and by the use of SQUID (a superconducting quantum interference device). Hydrogenation did not alter the crystal structure of the parent alloy, but it did increase the volume of the unit cell. An in situ Co K-edge XAS study of the hydride revealed that the valence state of Co increased during discharge (which is the release of hydrogen from the hydride). Hydrogenation of the parent alloy also reduced the magnetic moment. A possible mechanism of discharge for the hydride is also proposed.

Structural and magnetic properties of DyMn(2)D(6) synthesized under high deuterium pressure.

DyMn(2)D(6) has been prepared by applying high gaseous deuterium pressure on DyMn(2). This phase is isostructural with other RMn(2)D(6) (R = Y, Er) compounds and crystallizes with a K(2)PtCl(6) type structure having an ordered anion and a partially disordered cation arrangement because Dy and half the Mn atoms are randomly substituted in the same 8c site. The reverse susceptibility follows a Curie-Weiss law with an effective moment of 10 µ(B) similar to that of DyMn(2). Short range magnetic order, corresponding to ferromagnetic correlations, is observed in the neutron patterns up to 10 K and can be attributed to Dy-Dy interactions. The decomposition of the deuteride into Mn and DyD(2), studied by thermal gravimetric analysis, occurs between 470 and 650 K. A further deuterium desorption takes place above 920 K.

Cyclin D1 protein and CCND1 gene expression in colorectal cancer.

AIMS: To report the expression of cyclin D1 protein and its gene in a series of colorectal adenocarcinoma. METHODS: One hundred and eleven specimens of colorectal carcinomas and adjacent normal colorectal mucosa were investigated by staining with a monoclonal antibody against cyclin D1 and by RT-PCR. RESULTS: Expression of CCND1 gene was found in 54 out of 111 cases of colorectal cancers, while in normal mucosa the expression of this gene was not observed. Cyclin D1 protein expression was checked in the same group of adenocarcinoma cases. Presence of this protein was observed in 69 cases and for 43 of them also expression of its gene was found. Dependence between the presence of protein and the gene expression was statistically significant (p=0.0002). In the group of cases where CCND1 gene expression was detected, high level of its protein expression was found in 20 cases. The CCND1 gene expression was associated with metastases to lymph nodes (p=0.0181) and also with distant metastasis (p=0.0204). CONCLUSIONS: The combined measurement of both the gene and its protein product, is an important contribution to the study of molecular markers in histological material.

Usefulness of polyclonal antibodies raised against P65 oncofetal protein in immunohistochemical diagnosis of ductal breast cancer.

Paraffin-embedded infiltrating ductal breast cancer tissue slides (135) were analyzed by immunohistochemistry with the use of rabbit polyclonal anti-P65 oncofetal protein and mouse monoclonal anti-estrogen/progesterone receptor (ER, PR) antibodies. Analysis with anti-P65 antibody revealed the positive cytoplasmic reaction in 83 cases, 98 showed the nucleic reaction and 3 were immunologically negative. Among the analyzed cases 49 revealed both cytoplasmic and nucleic reactions. For the whole group of cancers the correlation was found between ER or PR level and P65 cytoplasmic reaction (r = 0.77 and 0.66, respectively) and low inverse correlation with nucleic localization of P65 protein. The percentage of positive cells with cytoplasmic expression of P65 was significantly higher in more histologically differentiated cancers (grade I and II according to Bloom and Richardson) than in grade III. Opposite tendency was observed for the nucleic expression of P65 protein. The percentage of immunopositive nuclei grew with the advance of the disease and was the highest in poorly-differentiated (grade III) tumors. The tumors with P65 cytoplasmic reaction were mainly small (T1, T2), without metastases to lymph nodes (N0) and distant metastases (M0). The dependence between P65 protein localization and clinical stage of disease (TNM classification) was evaluated statistically. The straight dependence existed between P65 nucleic reaction and tumor size (p = 0.0002), metastases to lymph nodes (p = 0.0032) and distant metastases (p = 0.0006). The obtained results suggest that the transfer of P65 protein from cytoplasm to nuclei of the breast cancer cells is connected with more clinically advanced stages and worse prognosis for the patients.

Uptake of radiolabeled morphiceptin and its analogs by experimental mammary adenocarcinoma: in vitro and in vivo studies.

Morphiceptin (Tyr-Pro-Phe-Pro-NH(2)) and its analogs modified at position 3: [D-Phe(3)]morphiceptin, [D-ClPhe(3)]morphiceptin and [D-Cl(2)Phe(3)]morphiceptin were synthesized and labeled with [(125)I] or [(131)I]. Their binding to membranes isolated from experimental adenocarcinoma was examined in vitro with the use of a cross-linking assay followed by the Western blot technique. The radioactive complex had molecular weight of about 65 kDa and was detectable by anti-mu-opioid receptor polyclonal antibody. Expression of the mu-opioid receptor in mouse mammary adenocarcinoma was confirmed by reverse transcriptase-polymerase chain reaction. The binding studies showed the highest affinity and capacity for [D-Phe(3)]morphiceptin (K(d) 0.39 and B(max) 1112) and [D-ClPhe(3)]morphiceptin (K(d) 1.8 and B(max) 220). Morphiceptin and its D-Cl(2)Phe analog had significantly lower B(max) values (131 and 83, respectively). Biodistribution experiments in tumor-bearing C3H/Bi mice with the use of the (131)I-labeled peptides confirmed the results of our in vitro studies. The highest accumulation of radioactive peptides in the tumor tissue was also found for peptides with D-Phe and D-ClPhe.

Occurrence of Podosphaera xanthii Race 2 on Cucumis melo in Brazil.

Powdery mildew is an important disease of melons (Cucumis melo L.) cultivated in greenhouses in Brazil. Currently, there are 5 races of Podosphaera xanthii (formerly known as Sphaerotheca fuliginea) and 2 races of Golovinomyces cichoracearum (formerly known as Erysiphe cichoracearum) described on melons worldwide, but only race 1 of P. xanthii has been reported in Brazil (1). However, typical whitish powdery fungal growth was observed on an experimental hybrid yellow melon resistant to race 1 of P. xanthii during the summer of 2000 in a greenhouse in Bragança Paulista, State of São Paulo. Conidia collected from diseased leaves were spread onto 0.5% water agar medium and maintained at 22°C for 24 h with 12 h of light and 12 h of darkness. Most of the germinated conidia displayed fibrosin inclusion bodies when observed in a solution of 3% potassium hydroxide (KOH), and approximately 1 of 50 also displayed forked germ tubes. These features allowed us to identify P. xanthii as the causal agent. Conidia raised on the susceptible yellow melon 'Amarelo CAC' were used to inoculate cotyledons of the differential melon lines (2) 'Hale's Best Jumbo' (susceptible to races 1, 2, and 3 of P. xanthii), 'PMR-45' (resistant to race 1 and susceptible to races 2 and 3), and 'PMR-6' (resistant to races 1 and 2 and susceptible to race 3). Inoculations were performed on 10 plants of each differential line and replicated four times. The presence or absence of symptoms was evaluated 18 days after inoculation. 'Hale's Best Jumbo' and 'PMR-45' were rated as susceptible while 'PMR-6' was rated as resistant, thus indicating the presence of race 2 of P. xanthii in Brazil. During field surveys from 2001 to 2003, this race was found on squash (Cucurbita moschata), summer squash (C. pepo), and melons in São Paulo. References: (1) F. J. B. Reifschneider et al. Plant Dis. 69:1069, 1985. (2) C. E. Thomas et al. Cucurbit Genet. Coop. 7:126, 1984.

p65 and c-erbB2 genes expression in breast tumors: comparison with some histological typing, grading and clinical staging.

Using PCR technique we have analyzed p65 and c-erbB2 genes expression in 47 frozen tissue slides taken from patients diagnosed as ductal and lobular breast cancer, classified as G3, and in a limited panel of proliferative breast disease cases. Expression of p65 was generally connected with small tumor size and with absence of metastases in regional lymph nodes. We have found interdependence between p65 gene expression and negative states of lymph nodes. On the contrary, c-erbB2 expression was observed in patients with large tumors and with metastases to the regional lymph nodes. Between both genes (p65 and c-erbB2) opposite interdependence was found. No statistical dependence between estrogen/progesterone receptor levels and p65 or c-erbB2 expression were noticed. The presence of p65 expression appeared in the group of proliferating breast disease cases which were connected with higher risk of breast cancer. Lack of p65 expression accompanied cases which were classified as fibroadenoma.

Expression of p65, DD3 and c-erbB2 genes in prostate cancer.

The expression of p65, DD3 and c-erbB2 genes was analyzed in 39 histologically verified human prostate cancers. The expression of p65 and DD3 genes was observed in significant percentage in well- and moderately-differentiated tumors. Both genes expression was lower in poorly differentiated tumors. On the contrary, c-erbB2 gene expression increased with advanced histological grading and reached the highest percentage in poorly-differentiated cancers. In the all investigated groups straight dependence between p65 and DD3 genes expression occurred. Opposite dependence was noticed in expression of p65/DD3 and c-erbB2 genes.

Immunohistochemical analysis of expression of a 65 kDa oncofetal protein (p65), epidermal growth factor receptor (EGFR), oncogene c-erb B2 and tumor suppressor gene p53 protein products in breast cancer patients.

Paraffin-embedded tissue slides from 88 infiltrating ductal breast carcinoma were examined by immunohistochemistry technique with the use of monoclonal antibody against human p65 antigen and polyclonal antibody against p65-like protein present in fetal bovine serum. Immunohistochemical analysis of expression of growth factor receptors (EGFR), protein product of oncogene c-erb B2 as well as protein product of mutated anti-oncogene p53 was also done. It was established that there is no correlation between p65 and c-erbB2, EGFR or p53 expression. In low differentiated tumors (grade III) high p53 index and high EGFR and c-erbB2 expression was connected with low p65 expression. The lack of c-erbB2 and EGFR and low p53 expression was combined usually with high p65 oncoprotein levels.

A 65 kDa oncofetal protein (p65), proliferating cell nuclear antigen (PCNA) and Ki67 expression in breast cancer patients.

Paraffin-embedded tissue slides from 89 infiltrating ductal breast carcinoma, 10 fibrocystic disease and 10 fibroadenoma were assessed immunohistochemically using monoclonal antibodies against human p65 antigen and polyclonal antibodies against p65-like protein present in fetal bovine serum. We did not find any evident differences in p65 detection by polyclonal and monoclonal antibodies, however, monoclonal antibody seems to be more specific. This factor is not induced by cellular proliferation associated with nonneoplastic diseases what was confirmed by immunohistochemical analysis of expression of p65 protein and well know markers of proliferation (proliferating cell nuclear antigen--PCNA and Ki67). It was established that there is no correlation between p65 and PCNA or Ki67 expression. High proliferating indexes (PI) for PCNA (PI-PCNA) or Ki67 (PI-Ki67) may help in selection of tumors with high proliferating activity independently from histological grade of malignancy established by routine methods. The estimation of p65 protein may be useful in the selection of precancerous changes and more differentiated ductal cancer of the breast what raises the possibility that p65 antigen may be helpful in the screening examination of women with high risk for cancer development.

Randomised trial of two regimens of chemotherapy in operable osteosarcoma: a study of the European Osteosarcoma Intergroup.

BACKGROUND: A previous trial by the European Osteosarcoma Intergroup (EOI) suggested that a short intensive chemotherapy regimen with doxorubicin and cisplatin might produce survival of operable, non-metastatic osteosarcoma similar to that obtained with complex and longer-duration drug regimens based on the widely used T10 multi-drug protocol. We undertook a randomised multicentre trial to compare these two approaches. METHODS: 407 patients with operable, non-metastatic osteosarcoma were randomly assigned the two-drug regimen (six cycles [18 weeks] of doxorubicin 25 mg/m2 on days 1-3 and cisplatin 100 mg/m2 on day 1) or a multi-drug regimen (preoperatively vincristine, high-dose methotrexate, and doxorubicin; postoperatively bleomycin, cyclophosphamide, dactinomycin, vincristine, methotrexate, doxorubicin, and cisplatin; this protocol took 44 weeks). Surgery was scheduled for week 9 for the two-drug group and week 7 for the multi-drug group. Analyses of survival and progression-free survival were by intention to treat. FINDINGS: Of 407 randomised patients, 391 were eligible and have been followed up for at least 4 years (median 5-6 years). Toxic effects were qualitatively similar with the two regimens. However, 188 (94%) of 199 patients completed the six cycles of two-drug treatment, whereas only 97 (51%) of 192 completed 18 or more of the 20 cycles of the multi-drug regimen. The proportion showing a good histopathological response (> 90% tumour necrosis) to preoperative chemotherapy was about 29% with both regimens and was strongly predictive of survival. Overall survival was 65% at 3 years and 55% at 5 years in both groups (hazard ratio 0.94 [95% CI 0.69-1.27]). Progression-free survival at 5 years was 44% in both groups (hazard ratio 1.01 [0.77-1.33]). INTERPRETATION: We found no difference in survival between the two-drug and multi-drug regimens in operable, non-metastatic osteosarcoma. The two-drug regimen is shorter in duration and better tolerated, and is therefore the preferred treatment. However, 5-year survival is still unsatisfactory and new approaches to treatment, such as dose intensification, are needed to improve results.

Expression of 65-kDa oncofetal protein in experimental hepatoma after anticancer therapy.

We have tested the expression of a 65-kDa oncofetal protein (p65) after combined treatment with menadione and methotrexate in hamsters transplanted with Kirkman-Robins hepatoma. The treatment of tumor-bearing animals with these compounds significantly inhibited both the tumor development and the expression of p65. This inhibition in tumor tissue was calculated from densitograms of Western blots. The inhibition of p65 expression was also confirmed in the serum of hepatoma bearing animals by using solid-phase radioimmunoassay (RIA) to quantify the specificity of polyclonal antibodies to fetal p65 molecules. Additionally, p65 was shown to localize both in cytoplasm and in the nuclear extracts prepared from hepatoma tissue.

Ki-1 large cell lymphoma: A clinical study of 12 cases.

The treatment and prognosis of Ki-1 positive lymphoma, a relatively new entity, remains largely unknown and the clinical features are still being defined. This is a retrospective analysis of our experience with 12 patients with Ki-1 lymphoma who were treated at our hospital over two years. Clinical presentation and management are described and an attempt is made to identify prognostic factors. The median age at presentation was 20 and the male/female ratio 1:1. Nine patients presented with nodal disease and three with extranodal. B symptoms were present in seven patients. Seven patients had stage I/II disease and five had stage IV. Immunophenotyping was available in 10 patients of which five were T-cell, two B-cell, and two null cell; one could not be categorized. All 12 patients received combination chemotherapy; five had consolidation radiotherapy. With a median follow-up of 11 months (range one to 29), actuarial survival was 67% at 29 months and disease-free survival 50% for all patients. Five out of seven patients with stage I/II and one out of five with stage IV remain disease free. We conclude that the clinical presentation is diverse and advanced stage appears to be the only identifiable adverse prognostic factor.

Malignant cystosarcoma phyllodes: A review of the clinical experience at King Faisal Specialist Hospital and Research Centre.

Twelve females with cystosarcoma phyllodes of the breast were referred to King Faisal Specialist Hospital and Research Centre (KFSH&RC) between 1980 and 1990, representing 0.8% of breast cancer patients seen during this period. Median age was 45 years (range 16 to 65 years). Seven patients (58%) were premenopausal. All patients presented with breast mass, which measured >10 cm in 58% of them. The median duration of symptoms was 15 months (range two to 174). Neither axillary lymph nodes nor distant metastases were seen at presentation. Surgery was the cornerstone of primary treatment (wide local excision in three and mastectomy in nine). Two patients received adjuvant chemotherapy and locoregional irradiation. All patients had malignant tumors histologically. Of the four tumors assayed for hormonal receptors, one was positive for estrogen and progesterone receptors. At a median follow-up of 17 months (range two to 77), four patients had relapsed; one died at 19 months and the projected five year survival is 83%. This limited data supports the observation that malignant cystosarcoma phyllodes is a distinctive clinicopathological entity of female breast cancer with a different natural history from carcinoma of the breast. Our survival results are similar to reported experience from the literature.