Kidney cell culture Macaca fascicularis as a candidate for vaccine development and in vitro model.

BACKGROUND: Cell culture is the proliferation of a cell population in vitro by isolating from the original tissue or growing from existing ones. One essential source is the monkey kidney cell cultures which have an essential role in biomedical study. This is due to the significant homology between the human and macaque genomes making these useful for cultivating human viruses, especially enteroviruses, and growing vaccines. METHODS: This study developed cell cultures derived from the kidney of Macaca fascicularis (Mf) and validated its gene expression. RESULTS: The primary cultures were successfully subcultured up to six passages, grew as monolayers, and exhibited epithelial-like morphology. The cultured cells remained heterogeneous in phenotype and they expressed CD155 and CD46 as viral receptors, cell morphology (CD24, endosialin, and vWF), proliferation, also apoptosis markers (Ki67 and p53). CONCLUSIONS: These results indicated that the cell cultures can be used as in vitro model cells for vaccine development and bioactive compound.

Increased expression of GAPDH in cynomolgus monkeys with spontaneous cognitive decline and amyloidopathy reminiscent of an Alzheimer's-type disease is reflected in the circulation.

Previous studies of aging cynomolgus monkeys from our group identified spontaneous age-associated cognitive declines associated with biomarkers and brain lesions reminiscent of Alzheimer's Disease (AD), in a proportion of aged monkeys. However, the molecular mechanisms that underlie the spontaneous amyloid disorders and cognitive declines observed in these affected monkeys have yet to be investigated in detail. Using reverse transcriptase quantitative real time PCR techniques, normalized to the ACTB housekeeping gene, we analyzed the expression patterns of a number of genes which have been implicated in amyloid and tau abnormalities, in well-characterized aged cynomolgus monkeys with cognitive decline. A significantly increased expression of the genes coding for glyceraldehyde 3-phosphate dehydrogenase (GAPDH), was found in aged-cognitive decline monkeys compared to age-matched healthy controls. GAPDH has been implicated in several neurodegenerative diseases and interacts with beta amyloid precursor proteins. These findings provide support for the utilization of cynomolgus macaques in translational preclinical research as valid spontaneous models in experimental investigations of the relationships among aging, cognitive decline, and the neuropathy of AD.

Imaging-based prostate cancer screening among BRCA mutation carriers-results from the first round of screening.

BACKGROUND: Male-carriers of BRCA1/2 gene mutations have an increased risk of prostate cancer (PCa) with a more aggressive phenotype. Current screening-guidelines suggest the use of prostate-specific antigen (PSA) only among BRCA2 carriers. Female carriers have extensive guidelines that include imaging. Our objective was to test the prevalence of PCa among BRCA carriers and examine screening strategies, using PSA and multiparametric magnetic resonance imaging (mpMRI). PATIENTS AND METHODS: We recruited men aged 40-70 years with BRCA1/2 germline mutations and no prior history of prostate biopsy. All men underwent an initial round of screening which included PSA, and prostate mpMRI. PSA was considered elevated using an age-stratified threshold of ≥1 ng/ml for 40-50 years of age, ≥2 ng/ml for 50-60 years of age, and 2.5 ng/ml for 60-70 years of age. Men with elevated PSA and/or suspicious lesion on mpMRI were offered a prostate biopsy. PSA levels, MRI findings, PCa incidence, and tumor characteristics were evaluated. Decision curve analysis was used to compare screening strategies. RESULTS: We recruited 188 men (108 BRCA1, 80 BRCA2), mean age 54 years (9.8). One hundred and ten (57%) had either elevated age-stratified PSA (75; 40%), a suspicious MRI lesion (67; 36%), or both (32; 17%). Of these, 92 (85%) agreed to perform a prostate biopsy. Sixteen (8.5%) were diagnosed with PCa; 44% of the tumors were classified as intermediate- or high-risk disease. mpMRI-based screening missed only one of the cancers (6%), while age-stratified PSA would have missed five (31%). Decision curve analysis showed that mpMRI screening, regardless of PSA, had the highest net benefit for PCa diagnosis, especially among men younger than 55 years of age. We found no difference in the risk of PCa between BRCA1 and BRCA2 (8.3% versus 8.7%, P = 0.91). Ninety percent had a Jewish founder mutation, thus the results cannot be generalized to all ethnic groups. CONCLUSIONS: PCa is prevalent among BRCA carriers. Age may affect screening strategy for PCa in this population. Young carriers could benefit from initial MRI screening. BRCA carriers aged older than 55 years should use PSA and be referred to mpMRI if elevated. TRIAL REGISTRATION: ClinicalTrial.gov ID: NCT02053805.

Detection and characterization of carbapenemase-producing Enterobacteriaceae in wounded Syrian patients admitted to hospitals in northern Israel.

Since 2013, four hospitals in northern Israel have been providing care for Syrian nationals, primarily those wounded in the ongoing civil war. We analyzed carbapenemase-producing Enterobacteriaceae (CPE) isolates obtained from these patients. Isolate identification was performed using the VITEK 2 system. Polymerase chain reaction (PCR) was performed for the presence of bla KPC, bla NDM, and bla OXA-48. Susceptibility testing and genotyping were performed on selected isolates. During the study period, 595 Syrian patients were hospitalized, most of them young men. Thirty-two confirmed CPE isolates were grown from cultures taken from 30 patients. All but five isolates were identified as Klebsiella pneumoniae and Escherichia coli. Nineteen isolates produced NDM and 13 produced OXA-48. Among a further 29 isolates tested, multilocus sequence typing (MLST) showed that ST278 and ST38 were the major sequence types among the NDM-producing K. pneumoniae and OXA-48-producing E. coli isolates, respectively. Most were resistant to all three carbapenems in use in Israel and to gentamicin, but susceptible to colistin and fosfomycin. The source for bacterial acquisition could not be determined; however, some patients admitted to different medical centers were found to carry the same sequence type. CPE containing bla NDM and bla OXA-48 were prevalent among Syrian wounded hospitalized patients in northern Israel. The finding of the same sequence type among patients at different medical centers implies a common, prehospital source for these patients. These findings have implications for public health throughout the region.

Temporal and spatial assessment of microbial communities in commercial silages from bunker silos.

Ensiling is a feed preservation method of moist forage crops that generally depends on naturally developing lactic acid bacteria to convert water-soluble carbohydrates into organic acids. While bacterial community dynamics have been previously assessed in bench-scale and pilot ensiling facilities, almost no studies have assessed the microbiomes of large-scale silage facilities. This study analyzed bacterial community composition in mature silage from bunker silos in three commercial production centers as related to pH, organic matter, volatile fatty acid composition, and spatial distribution within the ensiling bunker. It revealed significant physicochemical differences between "preserved" regions situated in the center and along the walls of the silage bunkers that were characterized by high concentrations of lactic acid and other volatiles and pH values below 5, and "spoiled" regions in the corners (shoulders) of the bunkers that had low lactic acid concentrations and high pH values. Preserved silage was dominated (>90 %) by lactic acid bacteria and characterized by high similarity and low taxonomic diversity, whereas spoiled silage had highly diverse microbiomes with low abundances of lactic acid bacteria (<5 %) that were sometimes characterized by high levels of Enterobacteriaceae. Spatial position had a much stronger impact on the microbial community composition than feedstock type, sampling date, or production center location supporting previous studies demonstrating that ecology and not geography is a major driver of environmental microbiomes.

An in vitro model to study interactions between Escherichia coli and lactic acid bacterial inoculants for silage in rumen fluid.

UNLABELLED: Previous studies have shown that silages treated with lactic acid bacteria (LAB) inoculants enhance ruminants' performance. The objective of the current experiments was to develop an in vitro model to study interactions between LAB silage inoculants and inoculated silages and Escherichia coli (EC) in rumen fluid (RF). Our hypothesis was that some inoculants inhibit EC in RF. For that purpose buffered RF was incubated under anaerobic conditions at 39°C with commercial strains of LAB silage inoculants or with laboratory corn and wheat silages treated with these LAB, an EC strain and with various ruminant feed ingredients. The EC strain was originally isolated from cattle manure and tagged with a plasmid expressing the green fluorescence protein and kanamycin and streptomycin resistance. Results indicate that the LAB or the treated silages did not suppress EC numbers in the RF. When the pH of the RF decreased below 5·0 the EC disappeared. We conclude that both LAB inoculants for silage and EC survived in RF for several days; however, the inoculants and silages treated with such inoculants did not inhibit EC in RF in vitro. SIGNIFICANCE AND IMPACT OF THE STUDY: Forage crops, silage and hay are initial stages of the food chain for humans. Cattle harbours and sheds enterobacteria regularly, some strains of which are pathogens. These can contaminate forage crops through field fertilization with cattle manure. The objective of this study was to develop an in vitro model to test whether lactic acid bacteria, which are used in silage inoculants, alone or in treated silages can inhibit Escherichia coli in rumen fluid. This study presents safety aspects and it is also part of a broad research effort aimed at finding out how LAB silage inoculants and inoculated silages enhance ruminant performance or exert probiotic effects in ruminants.

Spontaneous expression of the gene of KI67 and P53 in cynomolgus monkeys infected with papillomavirus.

Background and Aim: Cynomolgus monkeys (Macaca fascicularis) develop spontaneous infection of Papillomavirus (PV); thus, potentially beneficial for modeling human PV (HPV) infection study. Contrary to human origin, infection in cynomolgus monkeys does not always show evident clinical symptoms of cervical cancer. The absence of cervical cancer clinical symptoms leads us to investigate the molecular mechanism of the HPV infection in cynomolgus monkeys. This study aimed to investigate the messenger ribonucleic acid (mRNA) expression levels of KI67 and P53 genes, majorly known as biomarker oncogenesis of PV infection. Materials and Methods: The polymerase chain reaction (PCR) technique was used with MY11/MY09 primer to screen PV in cynomolgus monkey, further grouped as positive-PV and negative-PV infection groups. Real-time quantitative PCR was also applied to quantify the mRNA expression levels of KI67 and P53 genes in animals. Results: Increased expression of mRNA level of KI67 genes was significantly higher in Positive- PV group than negative-PV group. In contrast, the P53 mRNA expression level increased markedly higher in the negative-PV group than in the positive-PV group. Conclusion: Our study describes the potential of cynomolgus monkeys as a spontaneous oncogenesis model of PV infection-type. However, we used a limited number of cancer genetic markers. So, further study of other genetic markers is required to prove that cervical cancer could be developed naturally in cynomolgus monkeys.

Incidence of injuries and factors related to injuries in combat soldiers.

INTRODUCTION: Musculoskeletal injuries to the lower extremities are major factors contributing to drop out from military tasks. The aim of the present study was to determine the incidence of musculoskeletal injuries and the parameters that differentiate between the soldiers who incurred these injuries and those who did not along 14 weeks of an infantry commanders course. METHODS: One-hundred and sixty-eight participants were recruited from an infantry commanders course. The soldiers were tested before (pre), in the middle (middle) and at the end (last) of the course for anthropometric measurements, proprioceptive ability and dynamic postural balance (DPB), and filled out an ankle stability questionnaire (Cumberland Ankle Instability Tool (CAIT). A physiotherapist followed and recorded all musculoskeletal injuries incurred by the participants during the course. RESULTS: Fifty-eight participants out of the 168 (34.5%) reported some pain/injury. Time effects were found for body mass index, DPB asymmetry, DPB in posterior-medial (P-M) direction and proprioception ability. Injury effects were found for DPB asymmetry, DPB in P-M direction, CAIT and proprioception ability. An interaction was found for proprioception ability. The Cox regression showed that the variables that are mostly effecting injuries were pretesting proprioception ability, DPB asymmetry and CAIT. CONCLUSIONS: More than one out of three participants incurred musculoskeletal injuries, with deficits in proprioception ability, DPB and ankle stability in pretesting as major factors contributing to injuries. Further studies should look at the effect of specific exercises such as proprioception, DPB and ankle stability exercises for prevention and treatment of musculoskeletal injuries among combat soldiers.

Downregulation of peptide transporter genes in cell lines transformed with the highly oncogenic adenovirus 12.

The expression of class I major histocompatibility complex antigens on the surface of cells transformed by adenovirus 12 (Ad12) is generally very low, and correlates with the high oncogenicity of this virus. In primary embryonal fibroblasts from transgenic mice that express both endogenous H-2 genes and a miniature swine class I gene (PD1), Ad12-mediated transformation results in suppression of cell surface expression of all class I antigens. Although class I mRNA levels of PD1 and H-2Db are similar to those in nonvirally transformed cells, recognition of newly synthesized class I molecules by a panel of monoclonal antibodies is impaired, presumably as a result of inefficient assembly and transport of the class I molecules. Class I expression can be partially induced by culturing cells at 26 degrees C, or by coculture of cells with class I binding peptides at 37 degrees C. Analysis of steady state mRNA levels of the TAP1 and TAP2 transporter genes for Ad12-transformed cell lines revealed that they both are significantly reduced, TAP2 by about 100-fold and TAP1 by 5-10-fold. Reconstitution of PD1 and H-2Db, but not H-2Kb, expression is achieved in an Ad12-transformed cell line by stable transfection with a TAP2, but not a TAP1, expression construct. From these data it may be concluded that suppressed expression of peptide transporter genes, especially TAP2, in Ad12-transformed cells inhibits cell surface expression of class I molecules. The failure to fully reconstitute H-2Db and H-2Kb expression indicates that additional factors are involved in controlling class I gene expression in Ad12-transformed cells. Nevertheless, these results suggest that suppression of peptide transporter genes might be an important mechanism whereby virus-transformed cells escape immune recognition in vivo.

Interactions of Salmonella enterica with lettuce leaves.

AIMS: To investigate the interactions of Salmonella enterica with abiotic and plant surfaces and their effect on the tolerance of the pathogen to various stressors. METHODS AND RESULTS: Salmonella strains were tested for their ability to form biofilm in various growth media using a polystyrene plate model. Strong biofilm producers were found to attach better to intact Romaine lettuce leaf tissue compared to weak producers. Confocal microscopy and viable count studies revealed preferential attachment of Salmonella to cut-regions of the leaf after 2 h at 25 degrees C, but not for 18 h at 4 degrees C. Storage of intact lettuce pieces contaminated with Salmonella for 9 days at 4 degrees C resulted only in small changes in population size. Exposure of lettuce-associated Salmonella cells to acidic conditions (pH 3.0) revealed increased tolerance of the attached vs planktonic bacteria. CONCLUSIONS: Biofilm formation on polystyrene may provide a suitable model to predict the initial interaction of Salmonella with cut Romaine lettuce leaves. Association of the pathogen with lettuce leaves facilitates its persistence during storage and enhances its acid tolerance. SIGNIFICANCE AND IMPACT OF THE STUDY: Understanding the interactions between foodborne pathogens and lettuce might be useful in developing new approaches to prevent fresh produce-associated outbreaks.

Identification of a bovine mastitis Escherichia coli subset.

Eleven Escherichia coli isolates from clinical bovine mastitis cases (mastitic strains) and 11 from the cowshed environment (environmental strains) were compared, to determine if the former were a subset of the latter. The mastitic and environmental strains could not be distinguished according to O antigen and antibiotic sensitivity. All mastitic isolates showed significantly (P<0.0001) faster growth in milk and faster lactose fermentation than most (approximately 64%) environmental strains, but growth rates in nutrient broth did not differ. The rates of lactose fermentation and growth in milk were positively correlated. Adhesion and phagocytosis of mastitic strains by bovine PMN were significantly (P<0.0001) lower than those of environmental strains, and correlated negatively with growth in milk and lactose fermentation. The average percentages of killing by bovine leukocytes in the two sources were not statistically different. All mastitic strains were serum sensitive, whereas most ( approximately 72%) environmental ones were resistant. Finally, pulse-field gel electrophoresis revealed two main pulse type clusters, sharing a similarity coefficient of 79%. Cluster 1 comprised only environmental strains, whereas cluster 2 comprised mostly mastitic strains and only three environmental ones. Four mastitic strains shared a similarity coefficient of less than 74% with the other strains and were not included in the clusters. Our results suggest that clinical bovine mastitis E. coli isolates may form a subset of the general environmental E. coli population; they seem better able to multiply in the udder medium and to evade the host cellular innate immune response, and are genetically distinct from most environmental strains.

Interactions between Escherichia coli and the plant-parasitic nematode Meloidogyne javanica.

AIMS: To determine the potential of the plant-parasitic nematode Meloidogyne javanica to serve as a temporary reservoir for Escherichia coli. METHODS AND RESULTS: The adhesion to and persistence of E. coli on the surface of M. javanica were evaluated at different times and temperatures. A pure culture of green fluorescent protein (GFP) tagged E. coli was mixed with ca. 1000 J2 M. javanica for 2 h at 25 degrees C. The nematodes were then washed and the rate of the adhesion of the bacteria to the nematodes was determined by counting the viable nematode-associated E. coli, and by fluorescence microscopy. A dose-dependent adhesion rate was observed only at a bacterium to nematode ratio of 10(4)-10(6) : 1. The adhesion of E. coli to the nematodes was also tested over a 24 h-period at 4 degrees C, 25 degrees C and 37 degrees C. At 4 degrees C and 37 degrees C, maximal adhesion was observed at 5 h; whereas at 25 degrees C, maximal adherence was observed at 8 h. Survival experiments showed that the bacteria could be detected on the nematodes for up to 2 weeks when incubated at 4 degrees C and 25 degrees C, but not at 37 degrees C. CONCLUSIONS: Under laboratory conditions, at 4 degrees C and 25 degrees C, M. javanica could serve as a temporary vector for E. coli for up to 2 weeks. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings support the hypothesis that, in the presence of high concentrations of E. coli, M. javanica might serve as a potential vehicle for the transmission of food-borne pathogens.

Heparin cryoprecipitation reduces plasma levels of non-traditional risk factors for atherosclerosis in vitro.

AIMS: To show that heparin cryoprecipitation (HCP), an in vitro method of plasma purification, reduces the levels of in vivo modified proteins and non-traditional risk factors from plasma of atherosclerotic hemodialysis (HD) patients. METHODS: HCP was applied to plasma obtained from HD patients and controls, forming a precipitate--cryogel. Levels of fibrinogen, albumin, CRP, TNF-alpha, IL-6, advanced oxidation protein products, carbonylated fibrinogen and carbonylated albumin were determined in plasma before and after applying HCP and in the cryogel. RESULTS: Treatment of HD plasma with HCP, beyond the significant reduction of the increased levels of all the above-mentioned molecules, reduced fibrinogen, TNF-alpha, carbonylated fibrinogen and carbonylated albumin to control levels which were simultaneously found in the cryogel. CONCLUSIONS: HCP applied to plasma enables the simultaneous precipitation of modified molecules and circulating non-traditional risk factors for atherosclerosis. This study may serve as a base for the future development of a clinical purification technique.

The removal of 10/0 polyester (Mersilene) sutures after small incision congenital cataract surgery.

PURPOSE: To evaluate the use of 10/0 polyester (Mersilene) sutures for closure of small corneal incision after congenital cataract surgery. METHODS: The authors retrospectively reviewed the medical records of 58 cases (42 patients) who underwent congenital cataract extraction and intraocular lens implantation between 1999 and 2004, using Mersilene sutures. An examination looking for suture-related complications and retinoscopy was done 1 week after surgery and then every month for 6 months. The sutures were removed in cases of local tissue reaction, but not due to high postoperative astigmatism. Paired t-test was used to compare patients' age and astigmatism level in those cases who had suture removal (Group 1) as opposed to those who did not (Group 2). RESULTS: In 10 cases (17%) corneal vascularization, necessitating suture removal, was found during 6-month follow-up period, without the trigger of loose suture. Patient age was 3.5+/-3.3 years and 4.4+/3.3 years in Groups 1 and 2, respectively. At 1 week postoperatively the astigmatism value was 1.7+/-1.7 diopter (D) and 2.3+/-2.2 D in Groups 1 and 2, respectively, and it reduced to 0.9+/-0.8 in both groups at 6 months postoperatively. One case of endophthalmitis was encountered 2 days after suture removal. CONCLUSIONS: Removal of Mersilene sutures after congenital cataract surgery is required in cases of corneal vascularization, occurring during the first months postoperatively. Owing to the risk of general anesthesia and infection, suture removal should be considered with caution in cases of postoperative astigmatism.

The impact of temperature during the storage of table eggs on the viability of Salmonella enterica serovars Enteritidis and Virchow in the Eggs.

Salmonellosis is a foodborne infection of major economic importance. Contamination of table eggs with Salmonella, especially Salmonella enterica serovar Enteritidis, is a major health concern worldwide. Recently, S. enterica serovar Virchow has emerged as a major pathogen in Israel, where it is among the 3 most prevalent serovars found in poultry and the second most prevalent serovar isolated from individuals with salmonellosis. Although there is ample knowledge regarding the role of S. enterica serovar Enteritidis in contamination of eggs, virtually nothing is known regarding the possible association of S. enterica serovar Virchow with table eggs. Therefore, our goal was to examine the capability of serovar Virchow to contaminate chicken eggs. Commercial table eggs were inoculated independently with serovar Enteritidis and with serovar Virchow cells at a concentration of 10(5) cfu/egg, either on the shell surface or by injection into the yolk. The numbers of live Salmonella cells on the shell and within the egg were determined at various time points. At both low (6 degrees C) and room temperatures (25 degrees C), S. enterica serovar Virchow was not detected on the eggshell after 2 wk, whereas S. enterica serovar Enteritidis could be detected only sporadically at 25 degrees C. In contrast, within the eggs, S. enterica serovar Virchow survived for up to 6 wk at 6 degrees C, and it multiplied up to 10(9) cfu/mL of egg content from 2 to 8 wk postinoculation at 25 degrees C. In comparison, S. enterica serovar Enteritidis survived within the eggs up to 8 wk at 6 degrees C and at 25 degrees C. Our results suggest that in cold storage, serovar Virchow is able to persist for long periods (6 wk), and at room temperature, these bacteria can multiply within eggs and reach high concentrations. Therefore, eggs might be considered potential vectors for transmitting S. enterica serovar Virchow into the food chain.

Reversal of multidrug resistance in human colon cancer cells expressing the human MDR1 gene by liposomes in combination with monoclonal antibody or verapamil.

BACKGROUND: Colorectal cancer is a major cause of cancer-related mortality in the world and the second leading cause of neoplastic death in the United States. A major obstacle in the chemotherapy of this neoplasm is the emergence of multidrug resistance that is frequently associated with the expression of P-glycoprotein (p170) encoded by MDR1 (also known as PGY1) genes. Previously, we demonstrated that liposome-encapsulated doxorubicin is more cytotoxic than free doxorubicin in human promyelocytic leukemia and human breast cancer cells with the multidrug-resistant phenotype. PURPOSE: Our purpose was to investigate modulation of multidrug resistance by liposome-encapsulated vincristine (VCR) in a drug-resistant human colon cancer cell line HT-29mdr1 and the potentiation of this modulation in combination with monoclonal antibody MRK-16 or verapamil. METHODS: HT-29 parental cells and HT-29mdr1 cells were exposed to free VCR or liposome-encapsulated VCR alone or in combination with MRK-16 or verapamil. Cytotoxicity of cells after various treatments was determined by neutral red staining, and cellular content of VCR was measured by using radiolabeled VCR; p170 expression of cells was assessed by azidopine. RESULTS: HT-29mdr1 cells express a high amount of p170, thus conferring sixfold to sevenfold resistance to VCR compared with the parent cell line. Liposome-encapsulated VCR lowers drug resistance in HT-29mdr1 cells fourfold; IC50 values (concentration that causes 50% reduction in cell number) were 12.5 +/- 2.5 ng/mL compared with 42.5 +/- 5.0 ng/mL with free VCR. IC50 values for free VCR with empty liposomes were 25 +/- 1.25 ng/mL. The combination of MRK-16 and free VCR produced a twofold increase in cytotoxicity over free VCR in p170-expressing cells; the combination of MRK-16 and liposome-encapsulated VCR produced a 10-fold potentiation of cytotoxicity. toxicity. Nonspecific monoclonal antibody NR-LU-10 had no effect on cytotoxicity of HT-29mdr1 cells with free VCR or liposome-encapsulated VCR. The combination of 1.5 microM verapamil potentiated the cytotoxicity of free VCR ninefold to 10-fold, IC50 values reduced to 5.0 +/- 1.5 ng/mL, and in combination with liposome-encapsulated VCR, IC50 values reduced to 2.5 +/- 1.0 ng/mL, demonstrating a 15- to 17-fold potentiation of cytotoxicity. There were no significant differences in drug accumulation in HT-29mdr1 cells when treated with liposome-encapsulated VCR or free VCR. Liposomes inhibited the photoaffinity labeling of azidopine to p170 HT-29mdr1 cells. CONCLUSIONS: Liposome encapsulation of VCR effectively modulates multidrug resistance in human colon cancer cells and may become an important modality in treatment for colon cancers.

Preparation of multivesicular liposomes.

A novel type of liposome, named here multivesicular liposomes, was prepared by evaporation of organic solvents from chloroform-ether spherules suspended in water. Within each spherule were numerous water droplets that contained solutes to be trapped in liposomes upon solvent evaporation. Liposome preparations of different average diameters were made, varying from 29 +/- 10 microns to 5.6 +/- 1.7 microns. The liposomes were morphologically characterized by light microscopy and transmission electron microscopy. Materials successfully trapped within the liposomes ranged in molecular size from glucose to nucleic acids. Extremely high percentages of encapsulation (up to 89%) were achieved.

Peripheral polymorphonuclear leukocyte priming contributes to oxidative stress in early pregnancy.

OBJECTIVE: The cause of elevated blood leukocyte count in pregnancy is unknown. We hypothesized that priming of peripheral polymorphonuclear leukocytes (PMNL) caused this elevation. METHODS: Eleven women in the first trimester of pregnancy were included in this prospective study. Peripheral venous blood was drawn twice from each woman, before and after a medical abortion (pregnant and nonpregnant, respectively). Complete blood cell count, plasma alkaline phosphatase (ALP), and rate of superoxide release from separated phobrol 12-myristate-13-acetate (PMA)-stimulated PMNL were determined. RESULTS: The PMNL count in early pregnancy was significantly higher, with a significant increase in the PMNL rate of superoxide release compared to the nonpregnant state. A linear correlation between the rates of superoxide release and PMNL counts before and during pregnancy was found. ALP levels were significantly elevated in early pregnancy. CONCLUSION: The increased PMNL count is probably a compensatory response to PMNL priming. The increased rate of superoxide release from primed PMNL may contribute to oxidative stress in early pregnancy.

Arterial embolism in thyrotoxicosis with atrial fibrillation.

The incidence of atrial fibrillation (AF) in 142 patients with thryotoxicosis was 21% and that of thromboembolic disease was 8.5%. Male sex, increasing age, and associated rheumatic or hypertensive heart disease were significantly associated with the presence of AF. Thromboembolic episodes occurred in 12 (40%) of the patients with AF and in none of the 112 patients without AF. Cerebral emboli accounted for 53% of the episodes. In view of the grave prognosis of the embolic episodes, patients with thyrotoxicosis should be treated with heparin during attacks of AF and with oral anticoagulants when they have chronic fibrillation.