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The purpose of this study was to examine the hypothesis that fenceline contact between beef calves and cows at weaning reduces indices of behavioral distress and associated temporary reductions in weight gain. One hundred Angus/Hereford-cross calves were randomly assigned to five treatments for 7 d in each of 3 yr to determine the effect of different weaning techniques on their behavior and subsequent growth. Treatments were 1) fenceline separation from dams on pasture (F-P), 2) total separation from dams on pasture (S-P), 3) total separation from dams in a drylot (corral) preconditioned to hay (S-D-P), 4) total separation from dams in a drylot not preconditioned to hay (S-D-NP), and 5) nonweaned controls on pasture (C-P). At the end of the 7-d postweaning period, all calves were placed on pasture in large groups. Calves were weighed weekly for 10 wk. In the days following weaning, F-P and C-P calves spent more time eating (grazing or eating hay) than S-P and S-D-NP calves (P < 0.05). The S-P calves spent more time walking (pacing) than calves in the other four treatments (P < 0.05), which did not differ. The S-P calves also spent less time lying down than C-P, F-P, and S-D-P calves (P < 0.05); S-P and S-D-NP calves did not differ in lying time. The F-P calves vocalized less than S-P and S-D-NP calves (P < 0.05). In general, treatment differences were greatest during the first 3 d following weaning with d 2 (20 to 30 h after weaning) showing the greatest disparity. The F-P calves spent approximately 60% of their time within 3 m of the fence separating them from their dams during the first 2 d following weaning, whereas F-P cows spent about 40% of their time within 3 m of the fence during this period. Postweaning cumulative body weight gains of the F-P calves were greater than the gains recorded for the calves in the three totally separated treatments (which did not differ). The F-P calves gained 95% more weight than the average calf in the three totally separated treatments in the first 2 wk and were still heavier at 10 wk (21.4 vs 11.0 kg, respectively, at 2 wk and 50.0 vs 38.2 kg, respectively, at 10 wk; P < 0.05). It was concluded that providing fenceline contact between beef calves and cows for 7 d following weaning reduces behavioral indices of distress seen in the totally separated calves. In addition, fenceline contact with dams at weaning minimizes losses in weight gain in the days following separation. Totally separated calves did not compensate for these early losses in weight gain even after 10 wk.

Short-rib thoracic dystrophies (SRTDs) are congenital disorders due to defects in primary cilium function. SRTDs are recessively inherited with mutations identified in 14 genes to date (comprising 398 exons). Conventional mutation detection (usually by iterative Sanger sequencing) is inefficient and expensive, and often not undertaken. Whole exome massive parallel sequencing has been used to identify new genes for SRTD (WDR34, WDR60 and IFT172); however, the clinical utility of whole exome sequencing (WES) has not been established. WES was performed in 11 individuals with SRTDs. Compound heterozygous or homozygous mutations were identified in six confirmed SRTD genes in 10 individuals (IFT172, DYNC2H1, TTC21B, WDR60, WDR34 and NEK1), giving overall sensitivity of 90.9%. WES data from 993 unaffected individuals sequenced using similar technology showed two individuals with rare (minor allele frequency0.005) compound heterozygous variants of unknown significance in SRTD genes (specificity99%). Costs for consumables, laboratory processing and bioinformatic analysis wereAU$850 per sample. WES is sensitive, specific, efficient and cost-effective for mutation screening as well as gene discovery in SRTDs and can be considered a first-line methodology for mutation identification in affected individuals.

The rapid growth and widespread publicity about the World Wide Web has many implications for public reference librarians, but little is known about this group's attitudes towards the World Wide Web and the impact of the Web on reference work. Using face-to-face interviews, this study gathered data on the attitudes of reference librarians working within a large British Columbia public library consortium, focusing on how these information professionals feel about integrating the Web into their duties. The responses indicate that the librarians believe the Web has had a positive effect on their work with regard to efficiency and answer rates, but they still have serious concerns about accuracy of Web information, technical problems, and demands on them for customer training.

The disarmament agreements between the USA and Russia have resulted in huge quantities of fissile materials, plutonium and highly enriched uranium, becoming redundant. These materials are extremely attractive to terrorist organisations and rogue states who would like to add nuclear weapons to their armouries. Unfortunately, while the USA has done a great deal to assist Russia to protect its nuclear materials, European countries, including the UK, have not risen to the challenge of events. If chaos is to be avoided then Europe must provide Russia with far more assistance to help it deal with its nuclear inheritance. The problem highlights the importance of the activities of those who devote their lives to investigations of the best means of disposing of nuclear waste. The MOX route for disposing of plutonium is gaining support but it is still vitally important to have an alternative route available such as vitrifying a mixture of plutonium and fission products and burying it in a deep repository.

While in-vitro fertilization (IVF) was initially developed in women with tubal factor infertility, recent clinical studies have suggested that the presence of hydrosalpinges lowers implantation and pregnancy rates. We postulated that these hydrosalpinges cause impaired endometrial receptivity. A total of 103 women with hydrosalpinges were prospectively evaluated, and compared with 55 infertile and 44 fertile controls. All women had endometrial biopsies during the window of implantation, analysed by conventional histological criteria, and also stained for three integrin markers of endometrial receptivity (alpha1beta1, alpha4beta1 and alpha vbeta3). Women with hydrosalpinges (cases) expressed significantly less of the alpha vbeta3 integrin compared with controls. There was no difference in expression of alpha1beta1 or alpha4beta1 among groups. A significantly greater number of cases had out of phase histology and missing alpha vbeta3 (type I defects) and absent integrin expression despite normal histological maturation (type II) defects, compared with controls. Of 20 women with impaired endometrial receptivity who were also biopsied after hydrosalpinx surgery, 70% demonstrated increased alpha vbeta3 expression. Seventy-seven percent of type I and 57% of type II defects were corrected postoperatively. Using markers of endometrial receptivity, this study demonstrates that inflammatory hydrosalpinges have an adverse effect on endometrial receptivity, which in some cases may be overcome by surgical treatment of the hydrosalpinx.

Injection of extracts derived from adult caligid copepods induced a partial immunity to Lepeophtheirus salmonis in Atlantic salmon. Antigens were derived from the supernatant of adult lice extracts and were partially purified by Con A affinity chromatography. The antigens were also present in the pellet derived from adult lice extracts, Immunohistochemistry showed that antibodies present in the serum of rats immunized with the Con A purified extract bound predominantly to the gut of L. salmonis. Components of apparent sizes > 205 000, 165 000, 133 000, 130 000, 125 000, 114 000, 110 000, 96 000, 82 000, 78 000, 65 000, 46 000, 35 000, 31 000 and < 29 000 were present in the extract. Although the numbers of attached lice were initially higher on those fish in the immunized group, throughout the course of the challenge experiment the total numbers of lice did not vary greatly between the immunized or control groups of fish. Initially, an overall average of about 17 attached stages were recorded from each fish sampled and by the end of the experiment this figure had fallen to about five adult lice per fish. However, compared with control fish fewer gravid female lice were present on immunized fish and furthermore these lice possessed fewer eggs (P

Vaccination is potentially one of the most valuable methods of control against the caligid copepod Lepeophtheirus salmonis , the salmon louse, a major pathogen of farmed salmonids in the Northern hemisphere. A search for suitable potential antigens from this complex metazoan parasite forms the basis of this study. Salmon louse homogenate was separated by column chromatography into specific fractions which were assayed for a range of enzyme activities. Fractions found to be associated with enzymic activity or peaks of UV absorbance were used to raise antisera in 44 Wistar rats. An immunocytochemical investigation of the midgut of the adult louse using these specific antisera revealed immunostaining of the lumenal and basal surfaces of the louse gut. It is suggested that the proteins in the chromatographic fractions may represent digestive enzymes and/or structural or functional components of the salmon louse gut.

One of the remaining problems in the use of CdZnTe material as substrates in liquid phase epitaxy (LPE) of CdxHg1-xTe (CMT) layers is that of variation in lattice matching, i.e. Zn content, across substrates. This wil become increasingly important in the future as larger focal plane arrays of infrared detectors are required. The basic Bridgman growth process for CdTe/Cd0.96Zn0.04Te has been extended by applying the accelerated crucible rotation technique (ACRT). A marked reduction in axial Zn segregtion is seen in 50 mm diameter ACRT material, but this effect is smaller in the case of 75 mm diameter crystals. Radial variations in Zn content are small in both sizes of crystal, demonstrating the benefits obtained from ACRT stirring. Both macro- and microsegregation effects have been studied in these crystals in an attempt to understand the growth mechanism. Zinc distributions have been assessed by near-infrared transmission, X-ray lattice parameter measurements, atomic absorption spectrometry (AAS) and Auger electron spectrometry (AES). The last technique was used for the microsegregation studies, while AAS provides the absolute calibration for Zn content. Comparisons with segregation behaviour found in the literature will be given. It will be shown that the low temperature gradient and low growth rate lead to a degree of supercooling in the first-to-freeze region and this leads to significant Zn segregation in both radial and axial directions. As the crystals reach full diameter, the radial variation is decreased, presumably by the action of the ACRT, and axial segregation is also reduced.

A series of chain-extended poly(aryl ether ketones) (PAEKs) and poly(ether ketone ketone)/poly(ether ether ketone) (PEKK/PEEK) block copolymers have been prepared via the nucleophilic polycondensation and evaluated. The former were prepared by reacting a bisphenol, such as hydroquinone, with a series of difunctional PAEK oligomers, while the latter were prepared by reacting those same oligomers with a bisphenol, such as hydroquinone, and an activated dihalocompound, such as 4,4'-difluorobenzophenone. The chain-extended copolymers show good mechanical and thermal properties witch can be varied by varying the particular bisphenol used and/or the oligomer molecular weight

Amorphous polyarylates derived from 12–50 mol % of a t-butyl-substituted diacid, e.g., 5-t-butylisophthalic acid, exhibit miscibility with the amorphous phase of polyamides having aromatic and aliphatic character. Equally critical to blend miscibility is the aliphatic/aromatic carbon atom ratio (exclusive of the amide functionality) of the polyamide. An aliphatic/aromatic carbon atom ratio (α) greater than 1.4 but less than 2.5 is necessary. © 1992 John Wiley & Sons, Inc.

Objectives—A recent American study identified clinical factors which effectively predicted those patients who would have significant findings on cranial computed tomography. It was proposed to apply these criteria in a UK setting and to determine whether modifications could be made to improve their efficiency. Methods—A prospective observational study was conducted over a four month period including all non-trauma adult patients referred from the accident and emergency (A&E) department for urgent cranial computed tomography. Presenting symptoms and signs were analysed for ability to predict clinically significant computed tomography findings, namely: acute infarct, malignancy, acute hydrocephalus, intracranial haemorrhage, or intracranial infection. Results—Sixty two patients were included; 22 (35%) had significant findings on computed tomography. Applying the original criteria (any of: age 60 years or older, focal neurology, headache with nausea or vomiting, altered mental status) to the study population showed that no clinically significant tomograms would have been omitted but only 11% fewer performed. Modifying the criteria by removing "age 60 years or older" and replacing "altered mental status" with a Glasgow coma score

A total of 83 patients with metastatic transitional cell carcinoma who had previously received no systemic therapy entered a randomized phase II evaluation of carboplatin and cis-dichloro-trans-dihydroxy-bis-isopropylamine platinum IV (CHIP), administered respectively at 400 and 270mg./m.2 every 28 days. Among evaluable patients with measurable disease response rates were 3 of 22 (14%, 95% confidence interval 5 to 35%) for carboplatin and 4 of 25 (16%, 95% confidence interval 5 to 36%) for CHIP. Among 17 patients with evaluable but not measurable metastases (10 carboplatin and 7 CHIP recipients) there were no responses. Median survival for 64 evaluable patients was 4.8 months (5.0 months for carboplatin and 4.3 months for CHIP recipients). Independent factors prognostic for survival (p

As part of the Eastern Cooperative Oncology Group program for the assessment of new drugs in sarcomas, a Phase II trial of carboplatin was performed in patients who had received no more than one prior chemotherapy regimen. A total of 50 patients received either 400 mg/m2 or 320 mg/m2 depending on whether they had received prior radiotherapy. A response rate of 16% (95% confidence interval 6-32%) occurred in the 37 patients who had received doxorubicin as their only prior systemic therapy. Three of the six responses were complete and persisted for 7 to 34 months. In contrast none of the 13 patients who received carboplatin after initial progression on doxorubicin and subsequent progression on interferon alpha responded. The overall response rate was therefore 12% (95% confidence interval 5-24%). Toxicity was primarily hematologic, with 14 patients having Eastern Cooperative Oncology Group (ECOG) grade 3 toxicity and no grade 4 or 5 toxicities. In view of the number of complete responses, carboplatin should be studied further in untreated patients with advanced soft tissue sarcoma.

One of the most remarkable achievements in physical metallurgy occurred right at the start of the Civil Nuclear Reactor Programme when Alan Cottrell first predicted, then demonstrated, ‘irradiation’ or ‘Cottrell’ creep of uranium. This had immediate practical consequences because a Magnox reactor with uranium fuel was already operating at Calder Hall and it was found that the fuel elements were indeed deforming to an unacceptable extent owing to Cottrell creep. This deformation is a type of Newtonian flow, and other examples were found to occur in the fuel elements. In particular, the magnesium can components deformed by Nabarro–Herring creep; this was the first identification of this fundamental deformation mode occurring in an engineering component. Newtonian flow is also induced if a small load is applied to uranium while it is thermally cycled through its α–β phase change. This is a consequence of the volume changes that take place during the allotropic transformation. Volume changes also occ...

The crystal structure analysis of the title compound, C 9 H 10 IO 4 P, is a rare example of a full structural determination of an α-ketophosphonate ester, a type of compound which normally forms an oil. The P-C(carbonyl) bond is long, there is evidence that steric repulsion between the I and O(carbonyl) atoms is unexpectedly weak and there is a short intermolecular I...O contact linking the molecules in chains. The structure is compared with 2-iodobenzoyl compounds and with a nickel complex of an α-ketophosphonate.

Micromolar concentrations of the five-lipoxygenase inhibitor, MK 886 induce a "type 1" (apoptotic, extrinsic, death domain, receptor-dependent, caspase-positive) form of programmed cell death in Bcl-2-positive U937 human monoblastoid and HL-60 myeloid leukemia cells. A "type 2" (intrinsic, mitochondria-dependent, autophagic, in some examples caspase-negative (Panc-1)) form is induced in Panc-1 pancreatic and PC3 prostate cell lines. The latter two lines from epithelial-derived solid human cancers are Bcl-2-negative. Micromolar MK 886 induces an acute rise in Ca2+ in washed, Ca2+-poor U937 and HL60 cells in Ca2+ and Mg2+-free Hank's buffer. In U937 cells, much of the increase, or more properly redistribution, is nuclear in location (HL-60 not tested). No MK-886-induced acute Ca2+ increase developed in Panc-1 or PC3 cells. Bcl-2-positive HeLa cervical cancer cells exhibited an acute MK 886-induced increase in Ca2+. In the U937, PC3 and Panc-1 cells examined, MK-886 rapidly increased oxidative stress and decreased mitochondrial membrane potential, indicating that neither event is directly determinative for the altered distribution of Ca2+ or the form of PCD observed. Inhibition of increased U937 Ca2+ by the anti-oxidant, N-acetyl-L-cysteine, the effects of inhibitors of mitochondrial function including antimycin A, atractyloside, cyclosporin A, the L/N channel blocker loperamide, the intracellular chelator BAPTA and 2 agents, HA-14 and 3-methyl-antimycin A3 that impair Bcl-2 function further define these events. These differences in the Ca2+ response and possibly also the form of PCD that results may depend upon the presence of Bcl-2 or a related protein participating in a juxta-nuclear / nuclear Ca2+ ion channel. The role of mitochondria, the mechanism by which increased oxidative stress initiates the rapid release of Ca2+ from intracellular, possibly juxta-nuclear / nuclear sites or its redistribution to U937 Ca2+ nuclei, and whether this "signal" or possibly even ROS themselves mandate the type of PCD observed, presumably by differential modulation of transcription, remain to be determined. Lastly, these results demonstrate that, as might be expected, "soil" (cell type) trumps "seed" (inciting agent)".

MK 886, a 5-lipoxygenase inhibitor, induces a type 1 "apoptotic" form of programmed cell death in Bcl-2-positive U937 monoblastoid cells. In Ca2+-depleted, nonpermeabilized U937 cells studied with MK 886 in a Ca2+-free medium, an acute increase in Ca2+ occured within 10 to 20 seconds, detected with fura-2 measured with a spectrofluorimeter.The increased fluorescence was nuclear in location, as judged by confocal microscopy. The antioxidant, N-acetyl-L-cysteine, three agents that inhibit mitochondrialfunction at identified sites, antimycin A, atractyloside and cyclosporin A, the L/N-channel inhibitor, loperamide and BAPTA, an intracellular Ca+ chelator preloaded into cells each reduced the extent or prevented the acute MK 886-induced rise in Ca2+, as determined by radiometric detection. Rhodamine-2, a more selective mitochondrial Ca2+ probe, provided no evidence for nuclear Ca2+ originating from that extra-nuclear site or from the endoplasmic reticulum. With 2', 7'-dichloro-dihydrofluorescein-labelled cells to detect reactive oxygen species, MK 886 increased the initial fluorescent signal from a number of intracellular, largely extra-nuclear sites, including mitochondria. Two chemicals that inhibit the function of Bcl-2, HA14-1 and 2-methyl-antimycin A3, reduced the Ca2+ response to MK 886, if pre-incubated with the Bcl-2-positive U937 cells at 37 degrees C for several hours. MK 886 was previously shown to induce reactive oxygen species and a fall in mitochondrial membrane potential in both Bcl-2-positive U937 and in Bcl-2-negative PC-3 prostate and panc-1 pancreatic cancer cells. The latter solid tumor cells undergo an atypical "type 2" PCD without an acute rise in nuclear Ca2+.These results are consistent with an MK 886-induced increase of reactive oxygen species from intra-cellular sites including mitochondria which release Ca2+ located primarily at or near nuclei. These events may involve Bcl-2 participating in some form of Ca2+ channel and nuclear Ca2+ binding proteins undergoing conformational changes due to reactive oxygen species. Reasons for the different PCD responses in Bcl-2 positive lympho-hematopoietic compared to Bcl-2-negative solid cancer cell lines, respectively with and without the induced nuclear Ca2+ signal, remain to be defined.

The purpose of this study was to examine the hypothesis that fenceline contact between beef calves and cows at weaning reduces indices of behavioral distress and associated temporary reductions in weight gain. One hundred Angus/Hereford-cross calves were randomly assigned to five treatments for 7 d in each of 3 yr to determine the effect of different weaning techniques on their behavior and subsequent growth. Treatments were 1) fenceline separation from dams on pasture (F-P), 2) total separation from dams on pasture (S-P), 3) total separation from dams in a drylot (corral) preconditioned to hay (S-D-P), 4) total separation from dams in a drylot not preconditioned to hay (S-D-NP), and 5) nonweaned controls on pasture (C-P). At the end of the 7-d postweaning period, all calves were placed on pasture in large groups. Calves were weighed weekly for 10 wk. In the days following weaning, F-P and C-P calves spent more time eating (grazing or eating hay) than S-P and S-D-NP calves (P0.05). The S-P calves spent more time walking (pacing) than calves in the other four treatments (P0.05), which did not differ. The S-P calves also spent less time lying down than C-P, F-P, and S-D-P calves (P0.05); S-P and S-D-NP calves did not differ in lying time. The F-P calves vocalized less than S-P and S-D-NP calves (P0.05). In general, treatment differences were greatest during the first 3 d following weaning with d 2 (20 to 30 h after weaning) showing the greatest disparity. The F-P calves spent approximately 60% of their time within 3 m of the fence separating them from their dams during the first 2 d following weaning, whereas F-P cows spent about 40% of their time within 3 m of the fence during this period. Postweaning cumulative body weight gains of the F-P calves were greater than the gains recorded for the calves in the three totally separated treatments (which did not differ). The F-P calves gained 95% more weight than the average calf in the three totally separated treatments in the first 2 wk and were still heavier at 10 wk (21.4 vs 11.0 kg, respectively, at 2 wk and 50.0 vs 38.2 kg, respectively, at 10 wk; P0.05). It was concluded that providing fenceline contact between beef calves and cows for 7 d following weaning reduces behavioral indices of distress seen in the totally separated calves. In addition, fenceline contact with dams at weaning minimizes losses in weight gain in the days following separation. Totally separated calves did not compensate for these early losses in weight gain even after 10 wk.

OBJECTIVES: This research assessed the relationship between the deliveries of carcinogenic tobacco-specific nitrosamines (TSNAs) and the Federal Trade Commission (FTC) "tar" ratings of US commercial cigarettes. METHODS: Analysis of covariance (ANCOVA) was used to assess the explanatory power of FTC tar, the particular manufacturer, and other cigarette characteristics to predict the yields of four TSNAs (N'-nitrosonornicotine [NNN], 4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone [NNK], N'-nitrosoanatabine [NAT], and N'-nitrosoanabasine [NAB]) in 26 US commercial brands tested in the 1999 Massachusetts Benchmark Study. RESULTS: When FTC tar alone was used to predict TSNA yield, the squared correlation coefficient (R(2)) was only 38% for NNN, 76% for NNK, 46% for NAT, and 49% for NAB. Inclusion of manufacturer-specific variables significantly (p < 0.001) increased the estimated R(2) for three of the four species of nitrosamine to: 78% for NNN, 88% for NNK, and 81% for NAT. Inclusion of other cigarette characteristics (filter type, paper permeability, tobacco weight, tip dilution) did not reduce the significance of the manufacturer-specific effects. Federal Trade Commission nicotine and carbon monoxide (CO) yields were no better at predicting TSNA levels. CONCLUSIONS: FTC ratings for tar, nicotine, and carbon monoxide do not tell the entire story about the comparative yields of toxic agents in marketed cigarette brands. The significant manufacturer-specific effects suggest that proprietary blending and processing of tobacco matter as well. Public, brand-by-brand disclosure of the yields of TSNA and possibly other smoke constituents appears to be warranted.

The problem posed by the lack of response of cells in most solid cancers to current chemotherapy generally remains intractable.The use of cDNA arrays represents one global approach to identifying reasons for this failure. A messenger RNA response of pancreatic cancer (Panc-1) cells after culture for 24 hours with 12 microM cis-platinum was analyzed with a commercial cDNA array.Major drug-induced events included inhibition of messenger RNAs associated with cell proliferation and up-regulation of generally countervailing DNA repair, cellular stress, heat shock protein, glutathione stress-related and multiple drug resistance enzyme messenger RNAs, accompanied by a limited programmed cell death response.Induction of widespread normal stress-induced countervailing mRNAs by comparatively non-selective agents such as cis-platinum strongly biases against a successful therapeutic outcome. This paradoxical result of a therapeutic intent provides a further compelling argument for the use of specifically-targeted therapy such as growth factor receptor, tyrosine kinase and other discretely focused agents, probably employed in combinations based on expression of their targets in an individual patient's cancer, as identified by cDNA or proteonomic arrays.

The authors conducted a randomized Phase III trial of three treatment regimens for patients with residual, nonmeasurable, intra-abdominal metastatic disease after undergoing resection for primary colorectal carcinoma.To be eligible for this study, patients had to be both free of other malignancies and capable of starting their therapy within 3-6 weeks after surgery. They were required to have an Eastern Cooperative Oncology Group performance status3; to be chemotherapy, radiation, and immunotherapy naïve; to have adequate bone marrow, renal, and hepatic function; and to provide written, informed consent. The patients were divided into two cohorts: patients with no demonstrable hepatic metastasis (Group A) and patients with hepatic metastasis (Group B).The 229 patients in Group A were randomized to receive either 5-fluorouracil (5-FU) (n = 116 patients) or 5-FU with levamisole (n = 113 patients). The median survival (15.4 months and 15.3 months, respectively, for Groups A and B) was virtually identical. The two groups also were similar in terms of time to treatment progression, which was 7.9 months for group that received 5-FU alone 7.7 months for the group that received levamisole with 5-FU. The 168 patients in Group B with hepatic metastasis underwent a three-way randomization: 5-FU alone (n = 60 patients), 5-FU with levamisole (n = 54 patients), and 5-FU with hepatic irradiation (n = 54 patients). The median overall survival for the three treatment arms were similar, with 17.3 months for the group that received 5-FU alone, 16 months for the group that received 5-FU with levamisole, and 14.4 months for the group that received hepatic irradiation in addition to 5-FU: The time to treatment failure was 6.7 months, 6.8 months, and 8.3 months, respectively, for the three groups. The toxicity experienced by patients was as expected with the regimens, and no differences were observed between any of the treatment groups. The primary toxicities were hematologic and gastrointestinal. There was one treatment-related death due to adult respiratory distress syndrome, which occurred on the first day of the fourth cycle of 5-FU and levamisole. Other Grade 4 toxicities included nine patients with Grade 4 leukopenia, one patient with Grade 4 sepsis, and one patient with Grade 4 gastrointestinal toxicity, including blood loss and diarrhea.This study showed no treatment advantage for any of the combined modalities over 5-FU alone in this group of patients with intra-abdominal, nonmeasurable disease.

Metastases to the bony pelvis is an unusual pattern of the spread of primary pancreatic tumors. The authors report the presence of metastatic disease in the bony pelvis observed on bone scans in several patients who had been treated recently for pancreatic carcinoma. When bone scans that show metastatic disease in the pelvis are evaluated in patients with unknown primaries, the diagnosis of pancreatic carcinoma should be considered.

Pancreatic cancer kills nearly 29,000 people in the United States annually-as many people as are diagnosed with the disease. Chemotherapeutic treatment is ineffective in halting progression of the disease. Yet, specific immunity to pancreatic tumor cells in subjects with pancreatic cancer has been demonstrated repeatedly during the last 24 years. Attempts to expand and enhance tumor-specific immunity with biotherapy, however, have not met with success. The question remains, "Why can't specific immunity regulate pancreatic cancer growth?" The idea that tumor cells have evolved protective mechanisms against immunity was raised years ago and has recently been revisited by a number of research laboratories. In pancreatic cancer, soluble factors produced by and for the protection of the tumor environment have been detected and are often distributed to the victim's circulatory system where they may effect a more generalized immunosuppression. Yet the nature of these soluble factors remains controversial, since some also serve as tumor antigens that are recognized by the same T cells that may become inactivated by them. Unless the problem of tumor-derived immunosuppressive products is addressed directly through basic and translational research studies, successful biotherapeutic treatment for pancreatic cancer may not be forthcoming.

Incomplete programmed cell death is one explanation for the escape of cancer cells from therapy. Inhibitors of the enzyme 5-lipoxygenase reduce proliferation and initiate programmed cell death in many different types of malignantly transformed cells. The 5-lipoxygenase inhibitor, MK 886. induces an atypical form of programmed cell death in H-358 bronchiolar lung cancer cells. A genomic response of H-358 cells after 24 hr of culture at a 40 uM concentration that inhibited proliferation was analyzed with a Clontech human cDNA array containing 588 cDNAs corresponding to identified genes. The data grouped into 3 major categories and initial conclusions regarding countervailing, cellular stress, programmed cell death, DNA damage and repair mRNA-responses as possible reasons for escape from the antiproliferative response are discussed. The use of cDNA arrays to estimate the extent to which malignantly transformed cells respond to therapy or why they do not and so infer prognosis and identify possible therapeutic modifications is indicated.

The focal adhesion kinase (FAK) is a protein tyrosine kinase linked to signaling events between cells and the extracellular matrix. Studies at the Western blot level have demonstrated up-regulation of FAK expression in invasive breast and colon cancers. To assess p125FAK expression at the cellular level, we developed monoclonal antibodies that specifically detected FAK in formalin-fixed, paraffin-embedded tissue sections and analyzed the levels of FAK expression in human breast and colon tissues. Monoclonal antibody 4.47 demonstrated FAK-specific focal adhesion staining by immunofluorescence assays on BT-474 breast cancer cells and detected a Mr 125,000 protein by both Western blotting and immunoprecipitation analyses. Using immunohistochemical techniques, the expression of p125FAK was analyzed in 36 normal and 43 preinvasive or invasive human breast and colon tissues from individual patients. FAK was weakly expressed in most benign breast epithelium but was up-regulated at moderate or strong levels in 14 of 18 invasive breast carcinomas. In seven samples of ductal carcinoma-in situ, FAK was overexpressed. Borderline-to-weak expression of FAK was detected in the normal colonic epithelium. In the invasive colon cancers, FAK was overexpressed at moderate or strong levels in 13 of 15 tumors. Furthermore, FAK expression was up-regulated in areas of dysplastic, premalignant colon epithelium. These results provide the first evidence at the cellular level that FAK expression is variably overexpressed in breast and colon cancer and suggest that up-regulation occurs at an early stage of tumorigenesis.

We describe a genomic response of mRNAs associated with a subset of oncogenes, tumor suppressor and cell cycle-related genes in proliferating human Panc-1 pancreatic cancer cells after 24 hours of culture with MK886, a pleotrophic 5-lipoxygenase inhibitor. Ninety-eight of these cDNAs are represented in one of the sub-arrays included in the Clontech Human cDNA Expression Array. In this initial analysis, control cells exhibited apparent widespread low levels of disparate mRNA synthesis. In cells cultured with 40 microM MK886 for 24 hr, while most expressed genes, including a number of specific proliferation-enhancing genes such as c-myc were inhibited, 19 other ones including some countervailing genes including tyrosine SRC protein kinase, cyclins B1 and D1, CDC25B phosphatase and 40s ribosomal S19, amounting to 19 percent of the cDNAs resident on the chip were up-regulated at1.10 experimental/control values. Therapy-induced activation of compensatory proliferative genomic responses provides an additional explanation why malignant cells can fail therapy. Among their many future uses, gene chips clearly will be an extremely powerful tool for identifying relationships between the hierarchical linear and non-linear control and implementation-related cellular events and for identifying potential molecular targets tor cancer therapy.

We studied the relationship between current cigarette smoking and price among 34145 respondents, aged 15-29 years, to the 1992-1993 Tobacco Use Supplements to the Current Population Survey. The price elasticity of current smoking varied inversely with age: -0.831 (S.E. 0.402) for ages 15-17; -0.524 (S.E. 0.256) for ages 18-20; -0.370 (S.E. 0.188) for ages 21-23; -0.202 (S.E. 0.175) for ages 24-26; and -0.095 (S.E. 0.157) for ages 27-29. In response to higher prices, older youth were more likely to reduce the number of cigarettes smoked per day than to quit entirely. Among 15-17-year-olds, smoking cigarettes 'some days' was more sensitive to price than smoking 'every day'. Cigarette smoking was inversely related to the prices of premium brands, but not discount brands.

N-tertiary butyl-a-phenylnitrone, a free radical spin trap ator = 10 mM concentration, inhibited proliferation and reduced the viability of human pancreatic cancer (Panc-1) cells. The drug concentration determined the extent of inhibition, and with continued culture a proportion of the cells detached, most of which stained with trypan blue. Although hypodiploid cells were detected by flow cytometry of cells cultured with 20 mM NTBN, DNA laddering was absent and the TUNEL reaction negative. "Dark" cells present in samples cultured with 10 mM NTBN exhibited decreased cytoplasmic volume and increased staining with methylene blue and azure II, but lacked characteristic nuclear changes of type 1 programmed cell death. Cells cultured with10 mM of the spin trap exhibited nuclear and cytoplasmic changes more consistent with a non-type 1, type 2 variant of PCD with extensive cytoplasmic vacuolization. Careful analysis revealed evidence of marked pinocytosis in some cells. In view of the spin-trap associated pinocytosis, augmented uptake of chemotherapy in affected cells might be anticipated, but additive, synergistic or antagonistic interactions between NTBN and 5-fluorouracil were not observed.

Products of the arachidonic acid-metabolizing enzyme, 5-lipoxygenase, stimulate the growth of several cell types. Selective inhibitors of the enzyme, including SC41661A and MK886, reduce PC-3 prostate cell proliferation. With continued culture, cells die, but the mode of death, necrotic or nonnecrotic, has not been established.Flow cytometry, laddering after agarose electrophoresis of DNA from inhibitor-treated cells, and light and electron microscopy were employed to examine the type of death in PC-3 prostate cells cultured with either 5-lipoxygenase inhibitor.The inhibitors induced nonnecrotic, programmed cell death. SC41661A-treated cells exhibited "foamy," vacuolated cytoplasm and mitochondria with disrupted cristae and limiting membranes, while some cells contained numerous polysomes and extended hypertrophic Golgi and secretory cisternal networks. A proportion of the treated cells detached and the nuclei of these cells were characteristic of type 1 "apoptotic" programmed cell death. MK886, a 5-lipoxygenase-inhibitor with a different mechanism of action, induced nonnecrotic changes largely confined to the cytoplasm, most consistent with type 2 "autophagic" programmed cell death. In preliminary studies of mechanism, we demonstrated that PC-3 cells express mRNA for 5-lipoxygenase and for 5-lipoxygenase-activating protein. The less active inhibitor, SC45662 neither reduced proliferation nor induced DNA laddering. The antioxidant, N-acetyl-l-cysteine but not butylated hydroxy toluene or alpha tocopherol, partially reduced the inhibition of proliferation from SC41661A.SC41661A and MK886 inhibit PC-3 cell proliferation and induce a form of type 1 or type 2 programmed cell death, respectively. PC-3 cells contain messenger RNA for 5-lipoxygenase and 5-lipoxygenase-activating proteins. Drug-induced changes included altered redox potential, inferred from the increased survival due to the antioxidant and glutathione precursor, N-acetyl-l-cysteine. PC-3 cells are an appropriate model for studying the mechanism responsible for 5-lipoxygenase inhibitor-induced cellular suicide.

Crodamol PMP (polyoxypropylene (2) myristyl ether propionate) has been extensively employed for many years in a wide variety of human cosmetics in the absence of reported adverse reactions. The compound is being considered for use as a vehicle for veterinary pharmaceuticals in food-producing animals. This paper reviews information on the chemistry, metabolism, and toxicity of this compound, including the details of a recently completed multi-dose study in rats. Based upon this information, it is concluded that Crodamol PMP is a substance of apparently low toxic potential. Potential new uses of this compound must compare the likely exposure to the substance with the available toxicity information.

The 5-lipoxygenase inhibitors ETYA, SC41661A and MK886 reduced the proliferation and viability of Panc-1 human pancreatic cancer cells. The extent of inhibition depended upon drug concentration, and with continued culture, cells detached and stained with trypan blue. Although results from flow cytometry were those associated with programmed cell death, despite repeated attempts, no DNA laddering consistent with its later stages was detected, and studies with the TUNEL assay were negative. Light and electron microscopy of cells cultured with SC41661A provided morphologic evidence of a population of "dark" cells and of an incompletely expressed type 1 programmed cell death including margination of chromatin at the nuclear membrane and by consolidation and degeneration of cytoplasmic organelles, along with extensive vacuolization. Cells cultured with MK886 exhibited compact "dark" cells and an unusual cytoplasmic mode of cell death characterized by vacuolization and widely separated smooth internal membranes without diagnostic nuclear changes. This is in marked contrast to the extensive type 1 PCD induced by 5-lipoxygenase inhibitors cultured with human U937 monoblastoid cells. On balance, the response of Panc-1 cells to MK886 suggests expression of a variant type 2 (autophagic) cellular suicide, although some contribution from components of a "cytoplasmic" (type 3?) form of non-necrotic cell death may also be considered. In a European clinical trial, gamma linolenic acid, a polyunsaturated fatty acid that generates free radicals has been combined with 5-fluorouracil as chemotherapy for pancreatic cancer. Panc-1 cell proliferation was insensitive to inhibition by several chemotherapeutic agents employed clinically, including 5-fluorouracil, cisplatin or gemcitabine and only somewhat sensitive to GLA. When gamma linolenic acid was combined with MK886, the more effective of the two 5-lipoxygenase inhibitors, a synergistic reduction in Panc-1 cell number and viability occurred.

MK886 (Merck Frosst) is a selective in vivo inhibitor of 5-lipoxygenase, active at nanomolar concentrations. At micromolar concentrations, it inhibited the proliferation of U937 monoblastoid cells and of cultured malignant cells from patients with chronic myelogenous leukemia. These cells became morphologically apoptotic, a form of physiologic cell death. U937 cell apoptosis was assessed by flow cytometry, ultrastructure, DNA laddering and immuno-histology for free 3'OH-DNA. MK886-induced apoptosis developed over time as cells were recruited in concert with reduction in their numbers. Some CML cells exhibited cytoplasmic changes of apoptosis without typical nuclear changes. Under conditions used for measuring Ca2+ with Fura 2, 10 micromolar MK886 increased U937 intracellular Ca2+ 4-fold or more over the 8 minute period of measurement. Since MK886 inhibits the association of arachidonic acid with the 5-lipoxygenase activating protein, altered arachidonic acid metabolism may have contributed to these results.

Renal medullary carcinoma is a newly described, aggressive kidney tumor. All patients with the disease have been African-American with sickle cell (SC) trait or hemoglobin SC disease.Patient information was obtained from individual patient records and from the Department of Defense national data bank, The Defense Enrollment and Eligibility Reporting System. Data were obtained from either personal review of the patient's records or from discussion with the patient's physician. Cytogenetic studies were performed on one patient.Six patients are presented. All had SC trait. Median age was 24.5 years and 1 patient was female. Time from diagnosis to death averaged 3 months (range 1-7 mos). No objective responses were reported to a wide variety of chemo and immunotherapies: cyclophosphamide, doxorubicin, cisplatin; methotrexate, vinblastine, doxorubicin, and cisplatin; single agent interferon; single agent paclitaxel; or single agent vinblastine. Investigational regimens included topotecan, doxorubicin, and filgrastim; alpha-interferon, interleukin-2, and 5-fluorouracil; and single agent paclitaxel. Cytogenetic studies revealed numerous structural, as well as numerical anomalies. Of the cells successfully karyotyped (n=4), 2 contained abnormalities of chromosome 3 and all contained monosomy 11.Renal medullary carcinoma is an aggressive, chemoresistant tumor. Time from discovery of tumor to patient death is very short and has been altered by a wide variety of chemotherapies and immunotherapies. An unidentified genetic component is likely present.

A study was conducted to determine the antibiotic resistance and biochemical characteristics of 2690 Salmonella strains belonging to 52 serovars and isolated from environmental and feed samples from 270 turkey flocks in Canada. Resistance of the Salmonella strains to the aminoglycoside antibiotics varied widely; none of the strains were resistant to amikacin, 14.2% were resistant to neomycin, 25.8% were resistant to gentamicin, and 27.7% of the strains were resistant to kanamycin. Most strains (97.6%) were resistant to the aminocyclitol, spectinomycin. Regarding resistance to the beta-lactam antibiotics, 14.3% and 14.4% of the strains were resistant to ampicillin and carbenicillin, respectively, whereas only 5 (0.2%) of the strains were resistant to cephalothin. None of the strains were resistant to the fluoroquinolone ciprofloxacin or to polymyxin B. Resistance to chloramphenicol and nitrofurantoin was found in 2.4% and 7% of the strains, respectively. Only 1.7% of the strains were resistant to the trimethoprimsulfamethoxazole combination, whereas 58.1% were resistant to sulfisoxazole. Thirty-eight percent of the strains were resistant to tetracycline. Salmonella serovars differed markedly in their drug resistance profiles. Biochemical characterization of the Salmonella showed that the S. anatum, S. saintpaul and S. reading serovars could be divided into distinct biotypes.

This article reviews the published original sources of information on interactions of oral anticoagulants with dietary factors, points out deficiencies in our knowledge of these interactions, and suggests applications for this information in the clinical setting. As with many drug-nutrient interactions, the original references include a few experimental studies and many case reports. Deciding which interactions of oral anticoagulants with dietary factors are clinically relevant and determining the appropriate dietary prescription concerning each interaction involves, in most cases, an educated opinion rather than a conclusion based on extensive research. Enough information exists on the vitamin K content of foods and the quantity of vitamin K that alters coagulation status from the therapeutic range to provide the patient with advice concerning a group of foods to avoid and a group of foods to limit to one serving per day. With respect to other dietary factors that may interact with oral anticoagulants, the patient should be cautioned concerning supplements of vitamins A, E, and C and alcohol used chronically or ingested in large quantities.

We examined whether the arachidonic acid competitive antagonist, ETYA (5,8,11,14-eicosatetraynoic acid), modulated drug sensitivity in a cell line that over-expresses the multiple drug resistance protein, MDR1. ETYA was nontoxic to drug-sensitive parental KB3-1 cells or drug-resistant MDR KB8-5-11 cells, with an IC50 of 190 microM for both lines. ETYA (20 microM) increased rhodamine 123 accumulation in KB8-5-11 MDR cells but not in KB3-1 sensitive cells. Arachidonic acid at 20 microM did not alter rhodamine accumulation in either cell line. Increasing the concentration of ETYA from 40 to 160 microM or incubation beyond 30 min did not increase KB8-5-11 dye retention. Forty microM or more ETYA increased KB3-11 dye retention. In a 6 day proliferation assay of KB8-5-11 cells, a nontoxic (40 microM) concentration of ETYA reduced the IC50 for doxorubicin 4-fold, the IC50 for colchicine 2-fold, but had no effect on the IC50 for vinblastine. ETYA at 40 microM did not alter the IC50 for any drug tested with KB3-1 cells. Therefore: (a) ETYA (20 or 40 microM) modulated resistance of KB8-5-11 cells to several drugs to a limited extent, without potentiating toxicity in the parental line, while arachidonic acid did not. (b) Since cationic rhodamine 123 is concentrated in mitochondria, the extent is dependent upon the transmembrane potential, and increased dye retention due to ETYA may in part be related to altered ETYA-induced cell membrane potential.

We examined the effect of A63162 (Abbott), a selective inhibitor of 5-lipoxygenase on human prostate (PC3) cell proliferation. Within 5 min DNA synthesis was reversibly inhibited by 40 microM A63162, without altered cellular attachment or uptake of trypan blue. After 72 Hr, cells continued to be attached and exclude dye, were reduced in number and their histology was altered. Many treated cells were larger, more pleomorphic, with nuclear and cytoplasmic ultrastructural changes consistent with preparation for secretion. Some cells contained moderately swollen, distorted mitochondria. ETYA, a less selective inhibitor of 5-lipoxygenase that also inhibits cell replication, acutely reduced O2 uptake by 40%, but A63162 did not. The retention of the supravital mitochondrial dye, rhodamine 123 was increased by ETYA at 4 hr, but not after 24 hr; retention was not altered by A63162. Although the mechanism by which A63162 reversibly inhibits PC3 proliferation and initiates preparation for secretion is not identified, additional studies should further define its role in these events.

The sensitivity of cancer patient macrophages from different anatomical sites to arachidonic acid metabolism was investigated in tumor cell cytotoxicity assays. Alveolar macrophages and peripheral blood monocytes from 13 non-small cell lung cancer patients, peritoneal macrophages and peripheral blood monocytes from 13 ovarian cancer patients, and comparable macrophages from control patients with nonmalignant lung or gynecological diseases were tested. Inhibitors of either the cyclooxygenase pathway or the lipoxygenase pathway together with specific metabolites of each pathway were used to evaluate how these different macrophage populations are regulated by eicosanoids. In addition, metabolic studies were performed to compare directly the arachidonic acid metabolism of macrophages obtained from these different anatomical locations. The results demonstrate that the peripheral blood monocytes from lung cancer and ovarian cancer patients and the peritoneal macrophages from ovarian cancer patients are sensitive to cyclooxygenase inhibition; this was not seen with comparable macrophages from the relevant control patients. Sensitivity to modulation by cyclooxygenase inhibition correlated with increased cyclooxygenase metabolism and with the capacity of prostaglandin to mediate suppression of tumoricidal function in these populations of cancer patient macrophages. In contrast, alveolar macrophages from cancer patients were not sensitive to either cyclooxygenase inhibition or to prostaglandin-mediated suppression. No such differential influences were revealed for the lipoxygenase pathway of arachidonic acid metabolism in any macrophage population tested. Thus, eicosanoids, particularly those of the cyclooxygenase pathway, can be a critical immunoregulatory feature of certain tumor microenvironments.

Colonising bacteria detected within the follicular fluid of women undergoing assisted reproductive technology cycles has been associated with decreased embryo transfer rates and decreased pregnancy rates. The bacteria isolated from the follicular fluid of these women, at the time of trans-vaginal oocyte retrieval include Gram-positive anaerobic rods, Gram-negative anaerobic rods, streptococci, staphylococci, and lactobacilli. Some of these bacteria are opportunistic pathogens in the female genital tract. The expression of virulence factors including hyaluronidase and DNase, could affect the structural integrity of the oocyte and induce DNA fragmentation and apoptosis. Hyaluronin is a major carbohydrate component of the cumulus oocyte complex that could be a target for bacterial hyaluronidase. Other virulence factors associated with bacteria detected in cases of genital tract infections include the hydrolytic enzymes sialidase, β-galactosidase and β-N-acetylhexosaminidase (Howe 1999). Production of reactive oxygen species as a result of the presence of bacteria and bacterial heat shock proteins have been suggested as mechanisms responsible for increased DNA fragmentation and apoptotic progression of male gametes in colonised semen. Previous studies have not confirmed a direct relationship between sperm morphology and the level of sperm DNA fragmentation. DNA fragmentation in oocytes in relation to the presence of bacteria remains to be investigated. Oocyte quality is only determined by the assessment of cumulus cell morphology prior to in-vitro fertilisation. In-vitro testing of oocyte DNA integrity under various conditions may further the understanding of oocyte quality. In this study, mouse oocytes were used to investigate the structural and DNA integrity of oocytes after in vitro exposure to Lactobacillus spp., Streptococcus spp., and Staphylococcus spp. and determine whether the morphological appearance of the oocyte was predictive of the level of DNA fragmentation and whether DNA fragmentation can be attributed to certain bacterial species found colonising follicular fluid. (1) Howe L (1999). Mucinase and sialidase activity of the vaginal microflora: implications for the pathogenesis of preterm labour. International Journal of STD and AIDS 10(7), 442–446.

The capacity of alveolar macrophages and peripheral blood monocytes from patients with non-small cell lung cancer to develop tumoricidal function after in vitro stimulation with different macrophage activators was investigated. Alveolar macrophages were found to be impaired in their ability to develop cytotoxic activity compared with either the peripheral blood monocytes from the same patients or alveolar macrophages from patients with nonmalignant lung disorders. This result was observed consistently under diverse culture conditions and with different macrophage activators including gamma-interferon (gamma-IFN), granulocyte-macrophage colony-stimulating factor (GM-CSF), phorbol myristate acetate, or endotoxin. The impairment in tumoricidal function observed in alveolar macrophages was not associated with reduced target cell binding compared to peripheral blood monocytes. Alveolar macrophages from patients with lung cancer were found to secrete significantly greater amounts of tumor necrosis factor (TNF) and interleukin-1 (IL-1) than either peripheral blood monocytes from the same patients or alveolar macrophages from the patients with nonmalignant disorders. These results are consistent with either different regulatory pathways for cytotoxicity and cytokine secretion in the alveolar macrophages of patients with lung cancer or diversity in the subpopulations of cells responsible for these functions.