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.

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.

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.

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.

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.

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.

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.

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.

A systematic review of the effectiveness of Kinesio Taping--fact or fashion?

In this systematic review article, we assessed the effects of therapeutic Kinesio Taping (KT) on pain and disability in participants suffering from musculoskeletal, neurological and lymphatic pathologies. Four online databases (CINAHL, Cochrane Library, MEDLINE, PEDro) were comprehensively searched from their inception through March 2012. The initial literature search found 91 controlled trials. Following elimination procedures, 26 studies were fully screened. Subsequently, 12 met our inclusion criteria. The final 12 articles were subdivided according to the basic pathological disorders of the participants' musculoskeletal (N.=9), neurological (N.=1) and lymphatic (N.=2) systems. As to the effect on musculoskeletal disorders, moderate evidence was found supporting an immediate reduction in pain while wearing the KT. In 3 out of 6 studies, reduction of pain was superior to that of the comparison group. However, there is no support indicating any long-term effect. Additionally, no evidence was found connecting the KT application to elevated muscle strength or long-term improved range of movement. No evidence to support the effectiveness of KT for neurological conditions. As to lymphatic disorders, inconclusive evidence was reported. Although KT has been shown to be effective in aiding short-term pain, there is no firm evidence-based conclusion of the effectiveness of this application on the majority of movement disorders within a wide range of pathologic disabilities. More research is clearly needed.

The effect of cattle manure cultivation on moisture content and survival of Escherichia coli.

A new practice whereby wet slurry is added daily to the cattle manure bedding at the barn and cultivated has been developed in Israel. The objective of the present study was to examine the effect of manure cultivation on the persistence of Escherichia coli in a model system. A cow manure-derived E. coli strain was tagged with green fluorescence protein (GFP) and antibiotic resistance markers and was used to inoculate cow manure in 10-L buckets. After 3 successive cycles of inoculation and cultivation, wet slurry was added during an additional 2 cycles. After 32 d, the cultivated and noncultivated manure contained 677 ± 14 and 505 ± 2 g·kg(-1) DM, respectively. The cultivated manure remained drier compared with the noncultivated manure after the addition of wet slurry, and its texture remained lumpy compared with the compact, cohesive, and sticky texture of the noncultivated manure. Throughout the experiment, the counts of the tagged E. coli were less (P < 0.05) and disappeared faster in the cultivated than in the noncultivated manure. These results support the hypothesis that daily cultivation of manure may result in reduced incidence of mastitis and improves the welfare and performance of dairy cows.

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.

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.

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.

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.

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.

Fate of inoculated Escherichia coli in hay.

AIMS: To monitor the fate of inoculated Escherichia coli in dry and moist hay of various types, under laboratory conditions. METHODS AND RESULTS: Wheat, vetch and clover hay were used as received or wetted to 250-300 g kg(-1) moisture. The hay was inoculated at about 10(6) CFU g(-1) with a kanamycin-resistant E. coli strain that expresses the green fluorescence protein, and was stored in small open glass jars that were covered with aluminium foil. Three jars per treatment were sampled on days 1 and 3, or 4 and 7, or 8, 20 and 50, respectively, after the initiation of the experiments, and the numbers of E. coli in the hay were determined. The results indicated that E. coli disappeared from both dry and moist hay by 7-8 days after inoculation. However, in a few cases colonies that were presumed to be E. coli developed after incubation in Luria broth medium. CONCLUSIONS: The tagged E. coli strain usually disappeared rapidly from both the dry and the moist hay, in spite of the high level of inocula used. However, in some cases a few, possibly injured E. coli might have persisted, and could be detected after incubation in a rich growth medium. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is part of a risk assessment associated with sewage irrigation of forage crops in Israel. The results indicate that E. coli added to the hay is not likely to pose a health risk to cattle or to humans. Nevertheless, more research with natural strains of E. coli and other enteric pathogens that might be more adapted to forage conditions is warranted in order to ensure the safety of sewage-irrigated crops.

Astigmatism outcomes of scleral tunnel and clear corneal incisions for congenital cataract surgery.

PURPOSE: To evaluate astigmatism outcomes after congenital cataract surgery with intraocular lens implantation using clear corneal or scleral tunnel incisions. METHODS: We retrospectively reviewed the medical records of 46 children (67 eyes), aged 2 months to 12 years, who had undergone nontraumatic cataract extraction and intraocular lens implantation between 1996 and 2001, using a scleral tunnel incision (group 1), or a clear corneal incision (group 2). Refractive astigmatism was measured at 1 week, 3 months, and 5 months after surgery. Paired t-test was used to compare those variables, and Spearman's correlation was used to determine their relation to patient's age. RESULTS: Mean+/-SD astigmatism at 1 week postoperatively was 3.1+/-2.8 Diopter (D) and 2.1+/-1.7 D in groups 1 and 2, respectively. It significantly reduced to 1.1+/-1.2 D and 0.9+/-1.0 D, respectively, in the two groups at 5 months postoperatively (P<0.007). In both groups patients' age was significantly correlated with 1-week postoperative astigmatism (group 1: r=0.64; P=0.001; group 2: r=-0.58; P=0.003), and with the change in cylinder magnitude between 1 week and 3 months postoperatively (group 1: r=-0.67; P=0.001; group 2: r=0.50; P=0.013). CONCLUSION: Children who underwent congenital cataract surgery using clear corneal or scleral tunnel incisions showed high postoperative astigmatism at 1 week postoperatively, which spontaneously reduced during 5 months follow-up. Therefore, suture removal is not necessary in those cases.

Fate of Escherichia coli during ensiling of wheat and corn.

A recombinant Escherichia coli strain carrying a plasmid with an antibiotic resistance marker and expressing the green fluorescent protein was inoculated at a concentration of 3.8 x 10(8) CFU/g into direct-cut wheat (348 g of dry matter kg(-1)), wilted wheat (450 g of dry matter kg(-1)), and corn (375 g of dry matter kg(-1)). The forages were ensiled in mini-silos. The treatments included control (no E. coli added), application of tagged E. coli, and delayed sealing of the inoculated wheat. Three silos per treatment were sampled on predetermined dates, and the numbers of E. coli were determined on Chromocult TBX medium with or without kanamycin. Colonies presumptively identified as E. coli were also tested for fluorescence activity. Addition of E. coli at the time of ensiling resulted in a more rapid decrease in the pH but had almost no effect on the chemical composition of the final silages or their aerobic stability. E. coli disappeared from the silages when the pH decreased below 5.0. It persisted longer in silages of wilted wheat, in which the pH declined more slowly. Control silages of all crops also contained bacteria, presumptively identified as E. coli, that were resistant to the antibiotic, which suggests that some epiphytic strains are naturally resistant to antibiotics.