Acidic fibroblast growth factor (aFGF), also known as heparin-binding growth factor 1, is a mitogen for a variety of mesoderm- and neuroectoderm-derived cells. Several different aFGF mRNA species ...resulting from alternative splicing have been reported. These results suggest that the gene structure and regulatory mechanism for gene expression of aFGF are complex. As a first step toward understanding aFGF gene structure, we have isolated nine overlapping genomic DNA clones spanning 54 kbp and determined the complete DNA sequences of all three coding exons. Comparison of the nucleotide sequences between the human and bovine DNA showed that the sequence similarity extended 2400 bp downstream from the coding region. Cloning of the aFGF gene allowed us to characterize this locus in acute nonlymphocytic leukemia (ANLL) patients. A fraction of ANLL patients (10-20%) have a deletion in the long arm of chromosome 5, whose distal breakpoint overlaps the aFGF locus. Therefore, a prospective cohort of eight ANLL patients was screened using three different repetitive sequence-free probes derived from the aFGF locus. Using beta-globin gene as a normalization probe for hybridizing band intensities, we conclude that there is no allelic loss or gross rearrangement within the 40 kbp stretch of the aFGF gene locus in ANLL patients with or without 5q- deletion. Consistent with this observation, the aFGF mRNA was not detected in the mononuclear cells derived from either an ANLL patient or a normal individual as judged by the reverse transcription and polymerase chain reaction. We also identified a DNA fragment, 10.7 kbp upstream from the first coding exon of human aFGF, whose sequence is conserved in both the primate and rodent genomes. Further characterization of this fragment is likely to provide insight into the significance of this high degree of conservation.
and
are pathogens responsible for high larval oyster mortality rates in shellfish hatcheries. Bacteriophage therapy was evaluated to determine its potential to remediate these mortalities. Sixteen ...phages against
and
were isolated and characterized from Hawaiian seawater. Fourteen isolates were members of the
family, and two were members of the
In proof-of-principle trials, a cocktail of five phages reduced mortalities of larval Eastern oysters (
) and Pacific oysters (
) by up to 91% 6 days after challenge with lethal doses of
Larval survival depended on the oyster species, the quantities of phages and vibrios applied, and the species and strain of
A later-generation cocktail, designated VCP300, was formulated with three lytic phages subsequently named
phages vB_VcorM-GR7B, vB_VcorM-GR11A, and vB_VcorM-GR28A (abbreviated 7B, 11A, and 28A, respectively). Together, these three phages displayed host specificity toward eight
strains and a
strain. Larval
mortalities from
strains RE98 and OCN008 were significantly reduced by >90% (
< 0.0001) over 6 days with phage treatment compared to those of untreated controls. Genomic sequencing of phages 7B, 11A, and 28A revealed 207,758-, 194,800-, and 154,046-bp linear DNA genomes, respectively, with the latter showing 92% similarity to
phage YC, a strain from the Great Barrier Reef, Australia. Phage 7B and 11A genomes showed little similarity to phages in the NCBI database. This study demonstrates the promising potential for phage therapy to reduce larval oyster mortalities in oyster hatcheries.
Shellfish hatcheries encounter episodic outbreaks of larval oyster mortalities, jeopardizing the economic stability of hatcheries and the commercial shellfish industry. Shellfish pathogens like
and
have been recognized as major contributors of larval oyster mortalities in U.S. East and West Coast hatcheries for many years. This study isolated, identified, and characterized bacteriophages against these
species and demonstrated their ability to reduce mortalities from
in larval Pacific oysters and from both
and
in larval Eastern oysters. Phage therapy offers a promising approach for stimulating hatchery production to ensure the well-being of hatcheries and the commercial oyster trade.
The proto-oncogene c-N-ras frequently bears point mutations in ANLL cell DNA which endow it with the capacity to transform NIH/3T3 cells in vitro. Chronic myelogenous leukemia (CML) is a neoplasm ...highly related to ANLL since it involves the same hematopoietic progenitor cells and ultimately transforms to a neoplasm virtually indistinguishable from acute nonlymphoblastic leukemia (ANLL). Thus, we and others have examined ras genes in CML. This report confirms that ras gene activation is a very infrequent event in CML. However, a lymphoblastic cell line derived from a patient with CML did exhibit a novel second exon 61st codon activating mutation of c-N-ras.
Correspondence to Dr Jack Needleman, Fielding School of Public Health, UCLA, Los Angeles, CA 90095, USA; needlema@ucla.edu To the editors, In their editorial commenting on our paper ‘Association of ...registered nurse and nursing support staffing with inpatient hospital mortality’,1 Aiken and Sloane present our study results, conclusions and implications as if we examined the impact of substituting nursing support staff for professional nurses or registered nurses (RNs). What our paper and Griffiths’ paper examine are shortfalls from established levels of nurse support staffing as a complement to RN staffing within established unit-level nurse staffing models. Nurse staffing, nursing assistants and hospital mortality: retrospective longitudinal cohort study.
Activation of the cellular oncogene c-N-ras has been frequently observed in DNA from leukemic cells in acute myeloid leukemia (AML). Ras gene activation sufficient to mediate in vitro transformation ...and rodent tumorigenesis usually results from point mutations and amino acid substitutions in the 12th or 61st codons. In AML and the related myelodysplastic syndromes, amino acid substitution at the 13th codon has been observed. An activated c-N-ras gene from a 45-year-old patient with AML was isolated by transfection analysis and subjected to molecular cloning and sequence analysis. A point mutation of the 12th codon (GGT to GAT) resulting in aspartic acid substitution for glycine was observed. In other neoplasms such as colon cancer, specific ras mutations occur predominantly (e.g., K-ras, codon 12). This predominance has been of demonstrable value in analyzing large cohorts for ras activation with techniques that are rapid and economical, such as oligonucleotide hybridization. It had previously been thought that such a predominance for activation of c-N-ras at codon 13 existed in AML; however, this study in concert with others underscores the importance of 12th codon c-N-ras mutations, along with 13th and 61st codon mutations in the molecular pathogenesis of AML. Guanylate to adenylate transition mutations are commonly observed in AML and may provide insight into potential environmental leukemogens. Addressing all commonly prevalent ras activating mutations bears impact in the future design of molecular surveys of the role of ras activation in leukemogenesis.
The maternal and paternal genomes play different roles in mammalian brains as a result of genomic imprinting, an epigenetic regulation leading to differential expression of the parental alleles of ...some genes. Here we investigate genomic imprinting in the cerebellum using a newly developed Bayesian statistical model that provides unprecedented transcript-level resolution. We uncover 160 imprinted transcripts, including 41 novel and independently validated imprinted genes. Strikingly, many genes exhibit parentally biased--rather than monoallelic--expression, with different magnitudes according to age, organ, and brain region. Developmental changes in parental bias and overall gene expression are strongly correlated, suggesting combined roles in regulating gene dosage. Finally, brain-specific deletion of the paternal, but not maternal, allele of the paternally-biased Bcl-x, (Bcl2l1) results in loss of specific neuron types, supporting the functional significance of parental biases. These findings reveal the remarkable complexity of genomic imprinting, with important implications for understanding the normal and diseased brain.
OBJECTIVETo examine whether healthcare-associated infections (HAIs) and nurse staffing are associated using unit-level staffing data.
BACKGROUNDPrevious studies of the association between HAIs and ...nurse staffing are inconsistent and limited by methodological weaknesses.
METHODSCross-sectional data between 2007 and 2012 from a large urban hospital system were analyzed. HAIs were diagnosed using the Centers for Disease Control and Preventionʼs National Healthcare Safety Network definitions. We used Cox proportional-hazards regression model to examine the association of nurse staffing (2 days before HAI onset) with HAIs after adjusting for individual risks.
RESULTSFifteen percent of patient-days had 1 shift understaffed, defined as staffing below 80% of the unit median for a shift, and 6.2% had both day and night shifts understaffed. Patients on units with both shifts understaffed were significantly more likely to develop HAIs 2 days later.
CONCLUSIONSUnderstaffing is associated with increased risk of HAIs.
Using the NIH/3T3 cell transfection assay, activated cellular oncogenes have been detected in around 10% to 20% of human tumors. From a series of DNA preparations from tissues infiltrated with acute ...myelogenous leukemia (AML), 50% (3/6) caused transformation of NIH/3T3 cells. Thus AML appears to be the human tumor with the highest frequency of oncogenes detected by DNA transfection. In each case the oncogene involved was N-ras, a member of the ras gene family. Biologic and clinical parameters of AML patients with and without N-ras oncogenes in their tumors are discussed.
Background: A new type of computer-enhanced telemanipulator device for "robotic" laparoscopic surgery was recently approved. We prospectively evaluated the initial patients undergoing procedures with ...this new device at our institution. Methods: Patient demographics, operative indications, port placement, operative time, robot time, complications, and hospital stay were recorded. Follow-up evaluation was appropriate for the individual procedure. Results: Initially, 35 cases were managed. There were 22 anti-reflux procedures, 9 Heller myotomies, 1 pyloroplasty, 1 distal pancreatectomy with splenectomy, 1 esophagectomy with intrathoracic anastomosis, and 1 diagnostic laparoscopy. The operative times ranged from 88 to 458 min. The robot use times were between 16 and 185 min. There were no device-related complications. Conclusions: Computer-enhanced robotic telesurgery is a safe and effective treatment method for a variety of diseases of the proximal gastrointestinal tract. Further study is needed to determine the benefits of this approach as compared with current technology.
Few studies have focused on the significance of ras protein levels in human malignancy, in part because of the inherent difficulty in quantitation of the ras gene product. We have developed a method ...for the enzymatic determination of the ras gene product and have used this method for the quantitation of ras gene product levels in 19 patients with acute leukemia. This technique provides a practical means to assess p21 expression in leukemic cells ex vivo while avoiding the use of radioactive reagents. In addition, the mobility of the ras species of interest is determined. This assay should be easily modified for the use of other antibodies such as those reported to be specific for various ras species (i.e., H-, K- and N-ras), for specific ras mutations or for other nonras proteins. Because of the use of electrophoresis prior to quantitation of protein, the antibody used does not need to possess high specificity for the protein of interest.
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