Bevacizumab as adjuvant treatment of colon cancer: updated results from the S-AVANT phase III study by the GERCOR Group.

BACKGROUND: The bevacizumab-Avastin® adjuVANT (AVANT) study did not meet its primary end point of improving disease-free survival (DFS) with the addition of bevacizumab to oxaliplatin-based chemotherapy in stage III colon cancer (CC). We report here the long-term survival results (S-AVANT). PATIENTS AND METHODS: Patients with curatively resected stage III CC were randomly assigned to FOLFOX4, FOLFOX4-bevacizumab, or XELOX-bevacizumab. RESULTS: A total of 2867 patients were randomized: FOLFOX4: n = 955, FOLFOX4-bevacizumab: n = 960, XELOX-bevacizumab: n = 952. With a median of 6.73 years follow-up (interquartile range 5.51-10.54), 672 patients died, of whom 198 (20.7%), 250 (26.0%), and 224 (23.5%) were in the FOLFOX4, FOLFOX4-bevacizumab, and XELOX-bevacizumab arms, respectively. The 10-year overall survival (OS) rates were 74.6%, 67.2%, and 69.9%, (P = 0.003) and 5-year disease-free survival (DFS) rates were 73.2%, 68.5%, and 71.0% (P = 0.174), respectively. OS and DFS hazard ratios were 1.29 [95% confidence interval (CI) 1.07-1.55; P = 0.008] and 1.16 (95% CI 0.99-1.37; P = 0.063) for FOLFOX4-bevacizumab versus FOLFOX4 and 1.15 (95% CI 0.95-1.39; P = 0.147) and 1.1 (95% CI 0.93-1.29; P = 0.269) for XELOX-bevacizumab versus FOLFOX4, respectively. CC-related deaths (n = 542) occurred in 157 (79.3%) patients receiving FOLFOX4, 205 (82.0%) receiving FOLFOX4-bevacizumab, and 180 (80.4%) receiving XELOX-bevacizumab (P = 0.764), while non-CC-related deaths occurred in 41 (20.7%), 45 (18.0%), and 44 (19.6%) patients, respectively. Cardiovascular-related and sudden deaths during treatment or follow-up were reported in 13 (6.6%), 17 (6.8%), and 14 (6.3%) patients, in the FOLFOX4, FOLFOX4-bevacizuamb, and XELOX-bevacizumab arms, respectively (P = 0.789). Treatment arm, sex, age, histological differentiation, performance status, T/ N stages, and localization of primary tumor were independent prognostic factors of OS in stage III. CONCLUSIONS: S-AVANT confirms the initial AVANT report. No benefit of the bevacizumab addition to FOLFOX4 adjuvant therapy in patients with stage III CC was observed in terms of DFS with a negative effect in OS, without increase in non-CC related deaths. CLINICAL TRIAL IDENTIFICATION: NCT00112918.

Crop production in the USA is frequently limited by a lack of pollinators.

Most of the world's crops depend on pollinators, so declines in both managed and wild bees raise concerns about food security. However, the degree to which insect pollination is actually limiting current crop production is poorly understood, as is the role of wild species (as opposed to managed honeybees) in pollinating crops, particularly in intensive production areas. We established a nationwide study to assess the extent of pollinator limitation in seven crops at 131 locations situated across major crop-producing areas of the USA. We found that five out of seven crops showed evidence of pollinator limitation. Wild bees and honeybees provided comparable amounts of pollination for most crops, even in agriculturally intensive regions. We estimated the nationwide annual production value of wild pollinators to the seven crops we studied at over $1.5 billion; the value of wild bee pollination of all pollinator-dependent crops would be much greater. Our findings show that pollinator declines could translate directly into decreased yields or production for most of the crops studied, and that wild species contribute substantially to pollination of most study crops in major crop-producing regions.

Limited phenological and dietary overlap between bee communities in spring flowering crops and herbaceous enhancements.

Wild bee populations have undergone declines in recent years across much of the Western world, and these declines have the potential to limit yield in pollination-dependent crops. Highbush blueberry, Vaccinium corymbosum, and tart cherry, Prunus cerasus, are spring-blooming crops that rely on the movement of pollen by bees and other insects for pollination. Wild bee populations can be increased on farmland by providing floral resources, but whether the addition of these plants translates into increased pollinator density on crop flowers has not been documented in most cropping systems. To determine the importance of providing additional floral resources for wild bee pollinator communities, we selected blueberry fields and tart cherry orchards with and without herbaceous floral enhancements in western Michigan, USA. The bee communities visiting crop flowers, enhancements and control grassy field margins were sampled over a 5-yr period. In addition, the pollen diets of the most abundant wild bee crop pollinators were quantified across Michigan to better understand their foraging niches and to identify potentially important alternative host plants. The presence of floral enhancements did not increase the abundance of wild bees on either blueberry or cherry flowers during bloom. The bee community visiting blueberry was evenly composed of short-season bees that fly only during the spring and long-season bees that fly in both spring and summer. In contrast, the bee community visiting cherry was dominated by short-season spring bees. The majority of pollen collected by the wild bee communities visiting blueberry and cherry was from spring-flowering woody plants, with limited use of the herbaceous enhancements. Enhancements attracted greater abundance and species richness of bees compared to control areas, including twice as many floral specialists. Conserving summer-flying, grassland-associated bees is an appropriate goal for pollinator conservation programs. However, herbaceous enhancements may not provide adequate resources for the wild bees that pollinate spring-flowering crops. This study demonstrates that an examination of the pollen collected by wild bees across their flight periods can identify plant species to help them persist in intensively managed landscapes.

A survey of Australian and New Zealand clinical practice with neoadjuvant systemic therapy for breast cancer.

BACKGROUND: Neoadjuvant systemic therapy (NAST) has become an established treatment option for women with operable breast cancer. AIM: We aimed to better understand NAST treatment patterns, barriers and facilitators in Australia and New Zealand. METHODS: We undertook a cross-sectional survey of the current clinical practice of Australian and New Zealand breast cancer specialists. Questions included referral patterns for NAST, patient selection, logistics, decision making and barriers. RESULTS: Of 207 respondents, 162 (78%) reported routinely offering NAST to selected patients with operable breast cancer (median 9% of patients offered NAST). Specialty, location, practice type, gender or years of experience did not predict for offering NAST. In all, 45 and 58% wanted to increase the number of patients who receive NAST in routine care and in clinical trials respectively. Facilitators included the multidisciplinary team meeting and access to NAST clinical trials. Specialist-reported patient barriers included: patient desire for immediate surgery (63% rated as important/very important); lack of awareness of NAST (50%); concern about progression (43%) and disinterest in downstaging (32%). Forty-three per cent of participants experienced system-related barriers to the use of NAST, including other clinicians' lack of interest (27%); lack of clinical trials (24%) and unacceptable wait for a medical oncology appointment (37%). CONCLUSION: This group of Australian and New Zealand clinicians are interested in NAST for operable breast cancer in routine care and clinical trials. Patient- and system-related barriers that prevent the optimal uptake of this treatment approach will need to be systematically addressed if NAST is to become a more common approach.

Phase I ficlatuzumab monotherapy or with erlotinib for refractory advanced solid tumours and multiple myeloma.

BACKGROUND: Ficlatuzumab, a humanised hepatocyte growth factor (HGF) IgG1κ inhibitory monoclonal antibody, was evaluated for recommended phase II dose (RP2D), safety, pharmacokinetics (PKs), antidrug antibody (ADA), pharmacodynamics (PDs) and antitumour activity as monotherapy or combined with erlotinib. METHODS: Patients with solid tumours received ficlatuzumab 2, 5, 10 or 20 mg kg(-1) intravenously every 2 weeks (q2w). Additional patients were treated at the RP2D erlotinib. RESULTS: Forty-one patients enrolled at doses ⩽20 mg kg(-1). Common adverse events (AEs) included peripheral oedema, fatigue and nausea. Three patients experienced grade ⩾3 treatment-related hyperkalaemia/hypokalaemia, diarrhoea or fatigue. Best overall response (44%) was stable disease (SD); median duration was 5.5 months (0.4-18.7 months). One patient has been on therapy with SD for >4 years. Pharmacokinetics of ficlatuzumab showed low clearance (0.17-0.26 ml h(-1) kg(-1)), a half-life of 6.8-9.4 days and dose-proportional exposure. Ficlatuzumab/erlotinib had no impact on the PK of either agent. No ADAs were detected. Ficlatuzumab increased serum HGF levels. CONCLUSIONS: Recommended phase II dose is 20 mg kg(-1) q2w for ficlatuzumab monotherapy or with erlotinib. Preliminary antitumour activity and manageable AEs were observed. Pharmacokinetics were dose-proportional and consistent with other IgG therapeutics. Ficlatuzumab was not immunogenic, and serum HGF was a potential PD marker.

A national survey of obstetric early warning systems in the United Kingdom: five years on.

The Confidential Enquiries into Maternal Deaths in the UK have recommended obstetric early warning systems for early identification of clinical deterioration to reduce maternal morbidity and mortality. This survey explored early warning systems currently used by maternity units in the UK. An electronic questionnaire was sent to all 205 lead obstetric anaesthetists under the auspices of the Obstetric Anaesthetists' Association, generating 130 (63%) responses. All respondents reported use of an obstetric early warning system, compared with 19% in a similar survey in 2007. Respondents agreed that the six most important physiological parameters to record were respiratory rate, heart rate, temperature, systolic and diastolic blood pressure and oxygen saturation. One hundred and eighteen (91%) lead anaesthetists agreed that early warning systems helped to prevent obstetric morbidity. Staffing pressures were perceived as the greatest barrier to their use, and improved audit, education and training for healthcare professionals were identified as priority areas.

A randomized phase II study comparing capecitabine alone with capecitabine and oral cyclophosphamide in patients with advanced breast cancer-cyclox II.

BACKGROUND: Capecitabine and cyclophosphamide are active in patients with advanced breast cancer, have non-overlapping toxic effects and synergy pre-clinically. We explored the efficacy and toxic effect of an all-oral combination of capecitabine with cyclophosphamide versus capecitabine alone in a multicentre, randomized, phase II study. PATIENTS AND METHODS: Patients with locally advanced or metastatic breast cancer were randomized to treatment with capecitabine given continuously (666 mg/m(2) b.i.d. days 1-28) alone (C) or with oral cyclophosphamide (100 mg/m(2) days 1-14 of a 28-day cycle) (CCy) for up to six cycles. RESULTS: Eighty-two patients were randomized. There was no complete response. The proportions with partial response were 36% on C and 44% on CCy, a difference of 7.9% [95% confidence interval (CI) -13.4 to 29.1]. Significant toxic effect was uncommon: grade ≥3 diarrhoea in 4 (10%) versus 1 (3%) patients; grade ≥3 fatigue in 2 (5%) versus 5 patients (13%) and grade ≥2 hand-foot syndrome in 7 (17%) versus 11 (28%) patients receiving C versus CCy, respectively. Median progression-free survival was 3.1 months on C and 6.9 months on CCy, not significantly different statistically. There was no difference in overall survival. CONCLUSION: The difference in tumour response suggests a reasonable chance that CCy is superior to C alone.

Narrow pollen diets are associated with declining Midwestern bumble bee species.

Many species of bumble bee (Bombus) have declined in range and abundance across Europe, the Americas, and Asia, whereas other species have persisted and remain common and widespread. One explanation as to why some species have declined, based primarily on studies of the European bumble bee fauna, is that declining species have relatively narrow pollen-foraging niches and are less able to use alternative host plants in the absence of their preferred hosts. Though extensively explored in Europe, this hypothesis has not been investigated in North America, in part due to incomplete information on the foraging niche of many species. We selected 12 bumble bee species found in Michigan and quantified their pollen diets using museum specimens. We also extensively resurveyed the state to understand their contemporary status and distribution. Compared to a pre-2000 baseline, six species remain relatively common and widespread, whereas six species show range contractions of over 50%. There was a significant relationship between dietary breadth and distributional range change, with declined or declining species collecting around one-third fewer pollen types than stable species. Though there were significant compositional differences, we found no differences in the number of pollen types collected by species with differing tongue lengths. Overall, these results support the hypothesis that species with narrower dietary niches are at greater risk of decline. However, it is not clear if narrow dietary niches are a cause of declines, or if both are driven by an underlying factor such as proximity to the edge of climatic niches. Further research is needed to improve our understanding of dietary niche in bumble bees, and how it interacts with other factors to influence population trajectories of stable and at-risk species.

Design errors in vital sign charts used in consultant-led maternity units in the United Kingdom.

BACKGROUND: Paper-based charts remain the principal means of documenting the vital signs of hospitalised pregnant and postnatal women. However, poor chart design may contribute to both incorrect charting of data and clinical responses. We decided to identify design faults that might have an adverse clinical impact. METHODS: One hundred and twenty obstetric early warning charts and escalation protocols from consultant-led maternity units in the United Kingdom and the Channel Islands were analysed using an objective and systematic approach. We identified design errors that might impede their successful use (e.g. generate confusion regarding vital sign documentation, hamper the recognition of maternal deterioration, cause a failure of the early warning system or of any clinical response). RESULTS: We found 30% (n=36/120) of charts contained at least one design error with the potential to confuse staff, render the charts difficult to use or compromise patient safety. Amongst the most common areas were inadequate patient identification, poor use of colour, illogical weighting, poor alignment and labelling of axes, and the opportunity for staff to 'game' the escalation. CONCLUSIONS: We recommend the urgent development of an evidence-based, standardised obstetric observation chart, which integrates 'human factors' and user experience. It should have a clear layout and style, appropriate colour scheme, correct language and labelling, and the ability for vital signs to be documented accurately and quickly. It should incorporate a suitable early warning score to guide clinical management.

Evaluation of Nasonov Pheromone Dispensers for Pollinator Attraction in Apple, Blueberry, and Cherry.

Declines in the number of commercial honey bees (Apis mellifera L.) (Hymenoptera: Apidae) and some wild bee species around the world threaten fruit, nut, and vegetable production and have prompted interest in developing methods for gaining efficiencies in pollination services. One possible approach would be to deploy attractants within the target crop to increase the number of floral visits. In this study, we evaluate two new pollinator attractants, Polynate and SPLAT Bloom, for their ability to increase pollinator visitation and fruit set in apple (Malus pumila Mill.), highbush blueberry (Vaccinium sp. L.), and tart cherry (Prunus cerasus L.). Polynate is a plastic twin-tube dispenser loaded with a mixture of floral scent and Nasonov pheromone. SPLAT Bloom contains the same chemical formula as Polynate, but is applied as a 3 g wax dollop directly onto the tree or bush. The objectives of this study were to determine if Polynate and SPLAT Bloom increase the number of honey bee foragers and fruit set in apples, highbush blueberries, and tart cherries. We conducted replicated evaluations of 32 fields or orchards with and without putative attractants over three growing seasons. Both products failed to provide a measurable increase in pollinator visits or fruit set in these crops, indicating no return on investment for either product.

Borrelia burgdorferi bind to epithelial cell proteoglycans.

Borrelia burgdorferi adhere to mammalian cells in vitro but neither the ligand(s) nor the receptor(s) has (have) been clearly established. Using an in vitro attachment-inhibition assay, a B. burgdorferi attachment mechanism has been identified. Heparin, heparan sulfate, and dermatan sulfate reduced the attachment of virulent B. burgdorferi strain 297 to HeLa cells by approximately 60%. In addition, virulent, but not avirulent, B. burgdorferi strains B31, N40, and HB19 demonstrated heparin attachment-inhibition. Attachment to Chinese hamster ovary cells deficient in heparan sulfate proteoglycans was reduced by 68% compared to attachment to wild-type cells and was identical to attachment at maximum heparin inhibition to the wild-type cells. Pretreatment of HeLa cell monolayers with heparitinase, heparinase, and chondroitinase ABC, but not with chondroitinase AC, reduced borrelial attachment by approximately 50%. A moderately high affinity, low copy number, promiscuous B. burgdorferi glycosaminoglycan receptor was demonstrated by equilibrium binding studies. A 39-kD polypeptide, purified by heparin affinity chromatography from Triton X-100 extracts derived from virulent borrelia, was a candidate for this receptor. These studies indicate that one mode of B. burgdorferi attachment to eukaryotic cells is mediated by a borrelial glycosaminoglycan receptor attaching to surface-exposed proteoglycans on mammalian cells.

In the absence of other Fc receptors, Fc gamma RIIIA transmits a phagocytic signal that requires the cytoplasmic domain of its gamma subunit.

The transmembrane isoform of Fc gamma RIII, Fc gamma RIIIA, is found on NK cells, cultured monocytes, and tissue macrophages in association with a dimer of an accessory subunit, either gamma or zeta. Functions of individual Fc receptors have been difficult to analyze due to coexpression of the receptors on hematopoietic cells and permanent cell lines expressing Fc receptors. cDNAs for the alpha and gamma subunits of Fc gamma RIIIA were cotransfected into COS-1 cells, which lack endogenous Fc receptors, to evaluate receptor-mediated phagocytosis and changes in [Ca2+]i. Transfectants both bound and phagocytosed IgG-sensitized erythrocytes and, following activation of Fc gamma RIIIA, increased [Ca2+]i. The gamma subunit was essential both for the surface expression of the receptor and for transduction of the phagocytic signal. Truncation of the gamma subunit cytoplasmic domain (amino acids 65-80) eliminated phagocytic function. Phorbol ester inhibited phagocytosis in a concentration-dependent manner, but did not affect IgG-sensitized erythrocytes binding, suggesting that a protein kinase C-dependent pathway inhibits phagocytosis. The data indicate that a tyrosine containing cytoplasmic domain within the gamma subunit is required for phagocytosis by Fc gamma RIIIA.

Safety and efficacy of posaconazole in the long-term treatment of azole-refractory oropharyngeal and esophageal candidiasis in patients with HIV infection.

PURPOSE: To evaluate safety and efficacy of long-term posaconazole in HIV-infected patients with azole-refractory oropharyngeal candidiasis and/or esophageal candidiasis. METHOD: In this noncomparative, open-label study, participants received oral posaconazole 400 mg twice daily (bid) for 3 months. Enrolled patients (N = 100) included 60 from a previous 1-month acute study of posaconazole and 40 posaconazole-naïve participants. Participants with a clinical response could be followed untreated for up to 1 month afterwards. Participants who relapsed during follow-up, showed improvement at the end of 3 months of treatment (EOT), or were cured but likely to benefit from further therapy could continue on posaconazole 400 mg bid for up to 12 months. RESULTS: In the modified intent-to-treat population, clinical response (cure or improvement) occurred in 85.6% (77/90) at EOT. The results were similar in the previously treated participants and the posaconazole-naïve participants, 88.1% (52/59) and 80.6% (25/31), respectively. Posaconazole was well-tolerated, showing a similar safety profile during the 3-month study period and during suppressive therapy. The most frequently reported treatment-related adverse event was vomiting (4/100, 4%) during the early follow-up period (on or before day 105) and elevated hepatic enzymes (3/51, 6%) during the long-term follow-up (after day 105). CONCLUSION: Oral posaconazole 400 mg bid demonstrated long-term safety, tolerability, and efficacy, offering a long-term, suppressive treatment option for HIV-infected participants with azole-refractory mucosal candidiasis.

Comparison of female attractiveness and male response among populations of Choristoneura rosaceana (Harris) in western and eastern U.S. apple orchards.

Female obliquebanded leafrollers, Choristoneura rosaceana (Harris), collected from Oregon, Michigan, and New York were deployed in delta traps in Michigan apple orchards to compare their relative attractiveness to Michigan males of the same species. Females originating from Oregon attracted more males than females originating from New York during both generations of leafroller flight in Michigan. Also, females from Oregon attracted more males in Michigan than did "native" Michigan females during the first generation of flight. Analysis of gland extracts from the three populations revealed significantly more of each pheromone component in females originating from Oregon (approximately nine-fold more pheromone per female overall) than those from Michigan. However, there were no significant differences in the relative amounts of each pheromone component between Oregon and Michigan females. A 100:4:5:2 blend of Z11-14:OAc, E11-14:OAc, Z11-14:OH, and Z11-14:Ald was optimal for catching males in Michigan with no added or detrimental effect of Z11-14:Ald, confirming previous studies. However, 100:1 ratios of Z11-14:OAc relative to either E11-14:OAc or Z11-14:OH (also containing 2% Z11-14:Ald) captured more males in Oregon apple orchards compared with 100:4 and 100:10 ratios of Z11-14:OAc relative to either E11-14:OAc or Z11-14:OH. Addition of increasing amounts of Z11-14:Ald relative to Z11-14:OAc (range, 0-8:100) into a blend also containing 4% E11-14:OAc and 5% Z11-14:OH increased male catch in Oregon but not in Michigan. Our results suggest that pheromone blend quantity rather than blend quality may explain greater attractiveness of western compared with eastern female C. rosaceana to males in Michigan. Also, an optimized generic blend for monitoring male C. rosaceana across North America should contain Z11-14:Ald as has been previously shown, but should not exceed 4:100 ratios of both E11-14:OAc and Z11-14:OH relative to Z11-14:OAc for optimized catch of males in the western United States.

Vital signs and other observations used to detect deterioration in pregnant women: an analysis of vital sign charts in consultant-led UK maternity units.

BACKGROUND: Obstetric early warning systems are recommended for monitoring hospitalised pregnant and postnatal women. We decided to compare: (i) vital sign values used to define physiological normality; (ii) symptoms and signs used to escalate care; (iii) type of chart used; and (iv) presence of explicit instructions for escalating care. METHODS: One-hundred-and-twenty obstetric early warning charts and escalation protocols were obtained from consultant-led maternity units in the UK and Channel Islands. These data were extracted: values used to determine normality for each maternal vital sign; chart colour-coding; instructions following early warning system triggering; other criteria used as triggers. RESULTS: There was considerable variation in the charts, warning systems and escalation protocols. Of 120 charts, 89.2% used colour; 69.2% used colour-coded escalation systems. Forty-one (34.2%) systems required the calculation of weighted scores. Seventy-five discrete combinations of 'normal' vital sign ranges were found, the most common being: heart rate=50-99beats/min; respiratory rate=11-20breaths/min; blood pressure, systolic=100-149mmHg, diastolic ≤89mmHg; SpO2=95-100%; temperature=36.0-37.9°C; and Alert-Voice-Pain-Unresponsive assessment=Alert. Most charts (90.8%) provided instructions about who to contact following triggering, but only 41.7% gave instructions about subsequent observation frequency. CONCLUSION: The wide range of 'normal' vital sign values in different systems suggests a lack of equity in the processes for detecting deterioration and escalating care in hospitalised pregnant and postnatal women. Agreement regarding 'normal' vital sign ranges is urgently required and would assist the development of a standardised obstetric early warning system and chart.

Behavioral responses of Rhagoletis cingulata (Diptera: Tephritidae) to GF-120 insecticidal bait enhanced with ammonium acetate.

GF-120 is a baited formulation of the insecticide spinosad containing 1% ammonium acetate, developed for control of economically important fruit flies. The response of feral cherry fruit flies, Rhagoletis cingulata Loew, to GF-120 augmented with 0, 5, or 10% ammonium acetate was evaluated under orchard conditions. Significantly more flies were observed within 30 cm of bait droplets with 10% ammonium acetate added compared with standard bait or to a water control. These fly visits to GF-120 enhanced with 10 or 5% ammonium acetate lasted an average of 263.2 +/- 85.2 and 337.6 +/- 72.6 s, respectively, compared with 50.3 +/- 36.4 s for standard GF-120. Droplets containing additional ammonium acetate also were contacted by more flies, and more flies fed upon these droplets than on GF-120 or the water control. Furthermore, the duration of feeding on GF-120 bait enhanced with either level of additional ammonium acetate was significantly greater compared with standard GF-120 or water. Feeding events lasted between 61.5 +/- 30.7 and 73.4 +/- 21.0 s on enhanced GF-120 compared with 6.8 +/- 5.7 s on standard GF-120. Collectively, these results indicate that the interaction of feral R. cingulata with GF-120 droplets and the toxicant spinosad can be increased by addition of ammonium acetate.

Glycoprotein metabolism in inflammatory and neoplastic diseases of the human colon.

Carbohydrate compositions of the membrane and cytoplasmic fractions of human normal and cancerous colonic mucosa were compared in patients with blood groups O and B. The total sugar content in both fractions was reduced in the cancer tissues to about one-third of that in the normal colonic mucosa. The sugars that are associated with mucinous glycoproteins such as fucose and N-acetylgalactosamine were reduced significantly, while sugars that are primarily associated with "serum-type" glycoproteins were relatively unchanged or reduced to a lesser extent. The activities of glycoprotein:glycosyltransferases were variable, some showing so significant change, others beinb significnatly reduced in cancerous tissues. A polypeptidyl:N-acetylgalactosaminyltransferase (an enzyme that catalyzes the transfer of the first sugar to hydroxyamino acids of the protein core of mucinous glycoproteins), a sialyltransferase (involved in the addition of sialic acid to mucinous glycoproteins), and a galactoxyltransferase (thought to be responsible for blood group B antigenicity) were reduced in the cancerous colonic tissue. In contrast, the activities of these glycosyltransferases were unchanged in the colonic mucosa of patients with granulomatosis or ulcerative colitis. Glycosidase activities in the normal, cancerous, and inflammatory tissues were the same. These results suggest that in colonic cancer tissues the synthesis of one type of oligosaccharide chain may be greatly affected, while another family of oligosaccharides may remain relatively unaffected.

Different extracellular calcium requirements for proliferation of nonneoplastic, preneoplastic, and neoplastic mouse cells.

The DNA-synthetic and proliferative activities of freshly isolated, nontumorigenic C3H mouse skin cells (first passage) were lowest when the extracellular free (or ionic) calcium level was reduced to between 0.05 and 0.1 mM, whereas the extracellular free calcium level in cultures of repeatedly passaged, preneoplastic C3H/10T1/2 and MCA-C3H/10T1/2 type I mouse fetal fibroblasts had to be reduced to 0.01 mM or less before the DNA-synthetic and proliferative activities were minimal. This inhibition of DNA synthesis and cell multiplication by calcium deprivation was rapidly reversed by returning the extracellular calcium level to its normal value. In contrast, the neoplastic fibrosarcoma-forming, MCA-C3H/10T1/2 type III mouse fetal fibroblasts could synthesize DNA and could multiply indefinitely even in the presence of an extremely low concentration of extracellular free calcium. Thus, the extracellular calcium requirement for DNA synthesis and proliferation appears to reflect the tumorigenic potential of the cell.

Changes in cyclic adenosine monophosphate-responsive element binding proteins in rat hepatomas.

We applied Southwestern and Western blotting and gel retardation techniques to investigate the changes that occur in the cyclic adenosine monophosphate (cAMP)-responsive element (CRE) binding (CREB) proteins in rapidly growing, chemically induced 5123tc and 5123D Morris hepatomas. Using the CRE sequences from the c-fos, E2A, and somatostatin gene promoters, we identified in the nuclear proteins from normal unstimulated or proliferating rat liver cells six different protein factors of Mr 34,000, 36,000, 40,000, 47,000, 56,000, and 72,000 capable of binding to the element. The Mr 47,000 protein had the highest specificity for the core CRE, suggesting its importance in cAMP-mediated gene expression. We could not find the Mr 47,000 CREB protein in the 5123tc and 5123D hepatomas. Our efforts to detect this protein in the tumors by (a) using the CRE sequence from different gene promoters, (b) altering the protocol for extracting nuclear proteins, or (c) attempting to restore its DNA-binding property by phosphorylation [with endogenous protein kinase(s), a catalytic subunit of cAMP-dependent protein kinase, and protein kinase C/dephosphorylation (with alkaline phosphatase)] were unsuccessful. The loss of tje Mr 47,000 CREB protein from solid tumors of the Morris hepatoma is likely to be related to the neoplastic properties of the tumor cell rather than to cell growth because the level of this protein remained unchanged during a 6-day period of liver regeneration. The nuclear extract from the Morris hepatoma that did not have the Mr 47,000 CRE-binding factor contained proteins immunologically related to the CREB, c-Jun, and c-Fos proteins. We conclude that the Mr 47,000 factor represents a distinct member of the CRE-binding protein family and that its absence from the hepatomas may lead to aberrant expression of cAMP-inducible genes.

Regional and cellular localization of glycosyltransferases in rat small intestine. Changes in enzymes with differentiation of intestinal epithelial cells.

Optimal assay conditions were determined for four glycosyltransferases in rat small intestinal mucosal homogenates and the regional distribution and cellular localization of these enzymes was studied. For each glycosyltransferase, similar levels of activity were found in duodenal, proximal jejunal and distal ileal segments; activities of the galactosyltransferases were lower in the distal jejunal-proximal ileal segment. Planar section studies indicated that the undifferentiated crypt cells had significantly higher levels of sialyltransferase activities in the jejunum and ileum than the mature villus cells. A similar crypt to villus gradient was found for a galactosyltransferase in the ileum. These data suggest that glycoprotein synthesis may be active in the undifferentiated crypt cells and that certain glycosyltransferases may serve as marker enzymes for cellular differentiation in the intestine.