Genomic and transcriptomic comparison of Aspergillus oryzae strains: a case study in soy sauce koji fermentation.

The filamentous fungus Aspergillus oryzae is used in soy sauce koji making due to its high productivity of hydrolytic enzymes. In this study, we compared the genomes and transcriptomes of an industrial strain RD2 and a strain with decreased fermentation performance TS2, aiming to explain their phenotypic differences at the molecular level. Under the regulation of conidiation and fermentation conditions, the enhanced hydrolytic enzyme production and flavor precursor formation in RD2 described a complete expression profile necessary to maintain desirable fermentation performance. By contrast, central carbon metabolism was up-regulated in TS2 for fast growth, suggesting a conflicting relationship between mycelium growth and fermentation performance. Accumulation of mutations also lowered the fermentation performance of TS2. Our study has deepened the understanding of the metabolism and related regulatory mechanisms in desirable koji fermentation. A list of potential molecular markers identified here could facilitate targeted strain maintenance and improvement for better koji fermentation.

On equal footing: Trends in ankle/foot injuries for men vs. women.

OBJECTIVES: The objective of the current study was to examine trends in ankle/foot (A/F) injuries during the period 2001-2014, in order to determine whether the incidence of these injuries has changed and whether a previously identified difference in risk by gender still existed. In addition, other driver and crash-related risk factors were examined separately for men and women. METHODS: Passenger vehicle drivers aged 16+ were identified from NASS-CDS; weighted data were analyzed for model years 2001-2014. Model years (MY) were grouped as 2001-2004 (older) vs. 2005-2014 (newer), and drivers in frontal crashes were included. Ankle injuries included fractures and dislocations to the malleolus and distal tibia/fibula. Foot injuries included fractures and dislocations of the talus, calcaneus, and tarsal/metatarsal bones. Logistic regression models were constructed to identify risk factors, including MY, age, belt use, toepan/instrument panel intrusion, and body mass index (BMI) separately for each gender using odds ratios. RESULTS: The incidence of A/F injuries declined significantly between older and newer MY, especially for women. Whereas before MY 2005, ankle and foot injury risk was significantly higher for women than men, risks for ankle injury are now virtually the same for both genders, and women are only 1.2 times more likely than men to sustain a foot injury in a frontal crash. From multivariable regression models, however, it is apparent that there are different risk factors for A/F injuries for men vs. women. Body weight was a significant factor for both groups, but for men it was a risk only for those extremely obese, whereas for women those who were categorized as overweight were also at increased risk. Age greater than 55 was also found to be a risk factor for foot injuries among women but not men. For men and women, toepan intrusion remained the most important factor for both foot and ankle injuries, with significantly higher odds ratios noted for men. Foot pedals were a more likely injury source for women, whereas the toepan was more likely for men. In addition, belt use was protective for ankle injuries in women but not men. CONCLUSIONS: Significant declines in A/F injuries have been noted in recent years, especially for women, whose risks are now similar to those for men. However, significant risk factors remain for each gender, primarily related to body habitus (BMI) and toepan intrusion. Age was a risk factor for foot injuries among women, for whom the foot pedals were more likely to be an injury source. Toepan intrusion remains a major factor for both men and women, but, with the exception of 30+ cm of intrusion, odds ratios were primarily much higher for men in each category of intrusion.

Are frailty markers associated with serious thoracic and spinal injuries among motor vehicle crash occupants?

BACKGROUND: While age is a known risk factor in trauma, markers of frailty are growing in their use in the critically ill. Frailty markers may reflect underlying strength and function more than chronologic age, as many modern elderly patients are quite active. However, the optimal markers of frailty are unknown. METHODS: A retrospective review of The Crash Injury Research and Engineering Network (CIREN) database was performed over an 11-year period. Computed tomographic images were analyzed for multiple frailty markers, including sarcopenia determined by psoas muscle area, osteopenia determined by Hounsfield units (HU) of lumbar vertebrae, and vascular disease determined by aortic calcification. RESULTS: Overall, 202 patients were included in the review, with a mean age of 58.5 years. Median Injury Severity Score was 17. Sarcopenia was associated with severe thoracic injury (62.9% vs. 42.5%; p = 0.03). In multivariable analysis controlling for crash severity, sarcopenia remained associated with severe thoracic injury (p = 0.007) and osteopenia was associated with severe spine injury (p = 0.05). While age was not significant in either multivariable analysis, the association of sarcopenia and osteopenia with development of serious injury was more common with older age. CONCLUSIONS: Multiple markers of frailty were associated with severe injury. Frailty may more reflect underlying physiology and injury severity than age, although age is associated with frailty. LEVEL OF EVIDENCE: Prognostic and epidemiologic study, level IV.

The implementation and evaluation of an evidence-based statewide prehospital pain management protocol developed using the national prehospital evidence-based guideline model process for emergency medical services.

BACKGROUND: In 2008, the National Highway Traffic Safety Administration funded the development of a model process for the development and implementation of evidence-based guidelines (EBGs) for emergency medical services (EMS). We report on the implementation and evaluation of an evidence-based prehospital pain management protocol developed using this model process. METHODS: An evidence-based protocol for prehospital management of pain resulting from injuries and burns was reviewed by the Protocol Review Committee (PRC) of the Maryland Institute for Emergency Medical Services Systems (MIEMSS). The PRC recommended revisions to the Maryland protocol that reflected recommendations in the EBG: weight-based dosing and repeat dosing of morphine. A training curriculum was developed and implemented using Maryland's online Learning Management System and successfully accessed by 3,941 paramedics and 15,969 BLS providers. Field providers submitted electronic patient care reports to the MIEMSS statewide prehospital database. Inclusion criteria were injured or burned patients transported by Maryland ambulances to Maryland hospitals whose electronic patient care records included data for level of EMS provider training during a 12-month preimplementation period and a 12-month postimplementation period from September 2010 through March 2012. We compared the percentage of patients receiving pain scale assessments and morphine, as well as the dose of morphine administered and the use of naloxone as a rescue medication for opiate use, before and after the protocol change. RESULTS: No differences were seen in the percentage of patients who had a pain score documented or the percent of patients receiving morphine before and after the protocol change, but there was a significant increase in the total dose and dose in mg/kg administered per patient. During the postintervention phase, patients received an 18% higher total morphine dose and a 14.9% greater mg/kg dose. CONCLUSIONS: We demonstrated that the implementation of a revised statewide prehospital pain management protocol based on an EBG developed using the National Prehospital Evidence-based Guideline Model Process was associated with an increase in dosing of narcotic pain medication consistent with that recommended by the EBG. No differences were seen in the percentage of patients receiving opiate analgesia or in the documentation of pain scores.

Liver fatty acid-binding protein binds monoacylglycerol in vitro and in mouse liver cytosol.

Liver fatty acid-binding protein (LFABP; FABP1) is expressed both in liver and intestinal mucosa. Mice null for LFABP were recently shown to have altered metabolism of not only fatty acids but also monoacylglycerol, the two major products of dietary triacylglycerol hydrolysis (Lagakos, W. S., Gajda, A. M., Agellon, L., Binas, B., Choi, V., Mandap, B., Russnak, T., Zhou, Y. X., and Storch, J. (2011) Am. J. Physiol. Gastrointest. Liver Physiol. 300, G803-G814). Nevertheless, the binding and transport of monoacylglycerol (MG) by LFABP are uncertain, with conflicting reports in the literature as to whether this single chain amphiphile is in fact bound by LFABP. In the present studies, gel filtration chromatography of liver cytosol from LFABP(-/-) mice shows the absence of the low molecular weight peak of radiolabeled monoolein present in the fractions that contain LFABP in cytosol from wild type mice, indicating that LFABP binds sn-2 MG in vivo. Furthermore, solution-state NMR spectroscopy demonstrates two molecules of sn-2 monoolein bound in the LFABP binding pocket in positions similar to those found for oleate binding. Equilibrium binding affinities are ∼2-fold lower for MG compared with fatty acid. Finally, kinetic studies examining the transfer of a fluorescent MG analog show that the rate of transfer of MG is 7-fold faster from LFABP to phospholipid membranes than from membranes to membranes and occurs by an aqueous diffusion mechanism. These results provide strong support for monoacylglycerol as a physiological ligand for LFABP and further suggest that LFABP functions in the efficient intracellular transport of MG.

Causation and outcomes of diaphragmatic injuries in vehicular crashes.

BACKGROUND: This study aimed to identify vehicular and crash factors associated with diaphragmatic injuries (DIs). METHODS: DI presence was analyzed among Crash Injury Research and Engineering Network vehicular occupants (age ≥ 16 years) in relation to occupant and crash factors. Contact points (i.e., components in direct contact with occupants) involved in injury causation were analyzed. Cases with and without DI were compared in relation to mortality, Injury Severity Score (ISS) and the occurrence of injuries with a maximum Abbreviated Injury Scale (MAIS) score of 3+. Student's t test, Wilcoxon test, χ(2), and multiple logistic regression were used for statistical analysis. RESULTS: Of a total of 2,344 cases with complete data for analysis, 80 cases (3.4%) experienced DI. Multiple logistic regression analysis revealed a significant association with the occurrence of DI for near lateral impact (odds ratio, 7.71 [4.20-14.58]) and change in velocity (ΔV) of 40 km/h or greater (odds ratio 2.58 [1.29-5.24]). The seat belt and steering wheel were the most common contact points among frontal impact crashes, and the side interior surface and side hardware or armrest were the more common contact points among side impact crashes. DI cases experienced more MAIS3+ injuries in the head, chest and abdominal regions and exhibited higher median ISS (q1-q3) [42 (29-62.5) vs. 17 (10-27), p < 0.0001]. While mortality was higher among occupants with DI (48% vs. 14%), this effect disappeared when adjusted by ISS. CONCLUSION: DI are associated with lateral crashes and higher ΔV. Head, chest, and abdominal injuries occur more commonly among DI patients, and the presence of these injuries rather than the DI itself is responsible for the increased mortality in these patients. LEVEL OF EVIDENCE: Epidemiologic study, level III.

Changes in the Severity and Injury Sources of Thoracic Aorta Injuries due to Vehicular Crashes.

Research using the National Automotive Sampling System-Crashworthiness Data System (NASS-CDS) suggested a decreased adjusted risk of thoracic aorta injuries (TAI) for newer vehicles during near-side crashes and an increased adjusted TAI risk during frontal crashes. This study attempted to explore possible explanations of these findings. Adult front seat occupants in the Crash Injury Research and Engineering Network (CIREN) database through June 2012 were studied. TAI cases were compared with remaining cases in relation to crash and vehicular characteristics. TAI cases of later crash year (CY) (2004-2012) were compared to those in earlier CY (1996-2003) in relation to TAI severity (minor, moderate, severe and non-survivable). TAI cases in newer model year (MY) vehicles (1999-2012) were compared to those in older vehicles (1988-98) in relation to injury source (steering wheel, front, left, seat belt, air bag and other or unknown). Analysis was stratified by direction of impact (frontal and near-side) and the use of restraints. The similar TAI severity of earlier and later CY among frontal crashes suggests that the observed changes in the adjusted odds of injury seen in NASS-CDS are not due to an increase in injury detection. The decrease in TAI severity among newer vehicles in near-side crashes of later CY is consistent with a beneficial effect of crashworthiness improvements for this crash configuration. A shift of injury source in frontal crashes from the steering wheel in older vehicles to "front of vehicle structures", "seat belts" and "unknown and other" in newer vehicles should suggest potential sites for crashworthiness improvements.

Declining statewide trends in motor vehicle crashes and injury-related hospital admissions.

Numbers of crashes, rates of police-reported injury severity, and hospital admission rates were calculated for the ten year period between 2001 and 2010 in Maryland. Comparisons were made for two 5-year periods of 2001-2005 and 2006-2010. Crash characteristics remained similar for the two five-year periods, but there was a significant increase in occupant age. Declines in police-reported injury severity were noted for each of four age groups: 16-29, 30-54, 55-64, and 65+, with smaller declines among older occupants. In addition, there were significant declines in hospital admissions, comparing the two time periods. Although reductions in crashes may be attributable to various roadway, behavioral, and other safety improvement efforts, reductions in hospital admission rates most likely reflect major improvements in crashworthiness implemented during the past decade. For those admitted to hospitals, significant increases in injury severity were noted between the first and second time periods. There was an association between age and ISS, a measure of total bodily injury, with the highest ISS scores noted for the youngest and oldest groups (16-29 and 55+, respectively). In addition, there was a significant increase in the mean age over time, from 39 in 2001 to 43 in 2010, p<.001. In general, the incidence and severity of injuries increased for all body regions. There was also a significant increase in hospital mortality, although length of hospital stay remained the same. Given these trends, increased efforts need to focus on both injury prevention and treatment for the increasing population of older, sometimes frail, vehicle occupants.

Underreporting of driver alcohol involvement in United States police and hospital records: capture-recapture estimates.

This paper analyzes what portion of US nonfatal crashes are alcohol-involved and how well police and hospitals detect involvement. A capture recapture model estimated alcohol involvement from levels detected by police and hospitals and the extent of detection overlap. We analyzed 550,933 Crash Outcome Data Evaluation System driver records from 2006-2008 police crash report censuses probabilistically linked to hospital inpatient and emergency department (ED) discharge censuses for CT, KY (admissions only), MD, NE, NY, SC, and UT. We computed national estimates from NHTSA's General Estimates System.Nationally an estimated 7.5% of drivers in nonfatal crashes and 12.9% of nonfatal crashes were alcohol-involved. (Crashes often involve multiple drivers but rarely are two alcohol-involved.) Police correctly identified an estimated 32% of alcohol-involved drivers in non-fatal crashes including 48% in injury crashes. Excluding KY, police in the six states reported 47% of alcohol involvement for cases treated in EDs and released and 39% for admitted cases. In contrast, hospitals reported 28% of involvement for ED cases and 51% for admitted cases. Underreporting varied widely between states. Police reported alcohol involvement for 44% of those who hospitals reported were alcohol-involved, while hospitals reported alcohol involvement for 33% of those who police reported were alcohol-involved. Police alcohol reporting completeness rose with police-reported driver injury severity. At least one system reported 62% of alcohol involvement. Police and hospitals need to communicate better about alcohol involvement. Despite the proven effectiveness of brief alcohol intervention, EDs rarely detect, much less intervene with crash-involved drinking drivers. Both police and EDs particularly need to assess alcohol involvement in minor injury better.

Prescription medication usage and crash culpability in a population of injured drivers.

UNLABELLED: There has been increasing concern regarding the role of prescription drug use in the causation of traffic crashes. The goal of this research is to describe the prevalence of prescription drug use among injured trauma patients and determine the association between classes of drugs and crash culpability, a surrogate measure of crash risk. METHODS: Patient records, including chronic medication usage, for all drivers admitted to a trauma center following a traffic collision in 2008 (N=1,558) were linked with police crash reports to determine crash culpability. Multivariable analyses explored the association between medication use and crash culpability among non-drinking drivers. Adjusted odds ratios and 95% confidence intervals were compared among drivers who were and were not using central nervous system (CNS)-acting medications (single and multiple). RESULTS: 61.5% of all drivers were using any medications and usage increased with age, as did numbers of prescriptions per driver. Logistic regression analyses revealed that drivers who used CNS medications had an increased risk of culpability; those on more than one such medication had a crude (unadjusted) odds ratio of 2.16 for having caused the crash. Among drivers less than 45 years old, CNS medications did not significantly increase the risk of crash culpability. However, among drivers aged 45 or greater, the odds ratios for one, two, or 2+ CNS medications vs. none increased dramatically from 1.89 to 4.23 to 7.99, respectively. CONCLUSIONS: These results suggest that special attention should be given to older drivers (45+) using two or more CNS-acting agents.

Abca12-mediated lipid transport and Snap29-dependent trafficking of lamellar granules are crucial for epidermal morphogenesis in a zebrafish model of ichthyosis.

Zebrafish (Danio rerio) can serve as a model system to study heritable skin diseases. The skin is rapidly developed during the first 5-6 days of embryonic growth, accompanied by expression of skin-specific genes. Transmission electron microscopy (TEM) of wild-type zebrafish at day 5 reveals a two-cell-layer epidermis separated from the underlying collagenous stroma by a basement membrane with fully developed hemidesmosomes. Scanning electron microscopy (SEM) reveals an ordered surface contour of keratinocytes with discrete microridges. To gain insight into epidermal morphogenesis, we have employed morpholino-mediated knockdown of the abca12 and snap29 genes, which are crucial for secretion of lipids and intracellular trafficking of lamellar granules, respectively. Morpholinos, when placed on exon-intron junctions, were >90% effective in preventing the corresponding gene expression when injected into one- to four-cell-stage embryos. By day 3, TEM of abca12 morphants showed accumulation of lipid-containing electron-dense lamellar granules, whereas snap29 morphants showed the presence of apparently empty vesicles in the epidermis. Evaluation of epidermal morphogenesis by SEM revealed similar perturbations in both cases in the microridge architecture and the development of spicule-like protrusions on the surface of keratinocytes. These morphological findings are akin to epidermal changes in harlequin ichthyosis and CEDNIK syndrome, autosomal recessive keratinization disorders due to mutations in the ABCA12 and SNAP29 genes, respectively. The results indicate that interference of independent pathways involving lipid transport in the epidermis can result in phenotypically similar perturbations in epidermal morphogenesis, and that these fish mutants can serve as a model to study the pathomechanisms of these keratinization disorders.

Occupant and crash characteristics for case occupants with cervical spine injuries sustained in motor vehicle collisions.

BACKGROUND: Motor vehicle collisions (MVCs) are the leading cause of spine and spinal cord injuries in the United States. Traumatic cervical spine injuries (CSIs) result in significant morbidity and mortality. This study was designed to evaluate both the epidemiologic and biomechanical risk factors associated with CSI in MVCs by using a population-based database and to describe occupant and crashes characteristics for a subset of severe crashes in which a CSI was sustained as represented by the Crash Injury Research Engineering Network (CIREN) database. METHODS: Prospectively collected CIREN data from the eight centers were used to identify all case occupants between 1996 and November 2009. Case occupants older than 14 years and case vehicles of the four most common vehicle types were included. The National Automotive Sampling System's Crashworthiness Data System, a probability sample of all police-reported MVCs in the United States, was queried using the same inclusion criteria between 1997 and 2008. Cervical spinal cord and spinal column injuries were identified using Abbreviated Injury Scale (AIS) score codes. Data were abstracted on all case occupants, biomechanical crash characteristics, and injuries sustained. Univariate analysis was performed using a χ analysis. Logistic regression was used to identify significant risk factors in a multivariate analysis to control for confounding associations. RESULTS: CSIs were identified in 11.5% of CIREN case occupants. Case occupants aged 65 years or older and those occupants involved in rollover crashes were more likely to sustain a CSI. In univariate analysis of the subset of severe crashes represented by CIREN, the use of airbag and seat belt together (reference) were more protective than seat belt alone (odds ratio [OR]=1.73, 95% confidence interval [CI]=1.32-2.27) or the use of neither restraint system (OR=1.45, 95% CI=1.02-2.07). The most frequent injury sources in CIREN crashes were roof and its components (24.8%) and noncontact sources (15.5%). In multivariate analysis, age, rollover impact, and airbag-only restraint systems were associated with an increased odds of CSI. Using the population-based National Automotive Sampling System's Crashworthiness Data System data, 0.35% of occupants sustained a CSI. In univariate analysis, older age was noted to be a significant risk factor for CSI. Airbag-only restraint systems and both rollover and lateral crashes were also identified as risk factors for CSI. In addition, increasing delta v was highly associated with CSIs. In multivariate analysis, similar risk factors were noted. Of all the restraint systems, seat belt use without airbag deployment was found to be the most protective restraint system (OR=0.29, 95% CI=0.16-0.50), whereas airbag-only restraint was associated with the highest risk of CSI (OR=3.54, 95% CI=2.29-5.46). CONCLUSIONS: Despite advances in automotive safety, CSIs sustained in MVC continue to occur too often. Older case occupants are at an increased risk of CSI. Rollover crashes and severe crashes led to a much higher risk of CSI than other types and severity of MVCs. Seat belt use is very effective in preventing CSI, whereas airbag deployment may increase the risk of occupants sustaining a CSI. More protection for older occupants is needed and protection in both rollover and lateral crashes should remain a focus of the automotive industry. The design of airbag restraint systems should be evaluated so that they are not causative of serious injury. In addition, engineers should continue to focus on improving automotive design to minimize the risk of spinal injury to occupants in high severity crashes.

Pursuit of Licensure by Senior Drivers Referred by Police to a State Licensing Agency's Medical Advisory Board.

In all fifty United States and the District of Columbia, police Requests for Re-examination (RRE) concerning fitness to driver are accepted by licensing agencies. This study assessed licensing outcomes of senior drivers, ≥75 years of age, who had RREs submitted to the Medical Advisory Board (MAB) of a Maryland Motor Vehicle Administration from March 2005 through April 2007. RRE traffic event information (including crashed, did not crash), driver demographic information, initial MAB recommendations (suspension vs no suspension), driving occupational therapists assessments, and drivers' pursuit of continued licensure were entered into a database. During the period of study, 475 RREs were referred to the MAB. The percent of referred senior drivers (n=240, 50.4%) was similar to that of younger drivers (n=235, 49.5%). A higher percentage of senior drivers retired from driving compared to younger drivers; being, 57.1% vs 23.8% (p <.01), respectively. Further analyses limited to the 240 senior drivers found: 139 (57.9%) were men, 150 (62.5%) were 75-84 years of age, 119 (49.5%) were noted to be disoriented at the traffic scene, 141 (58.8%) were involved in a crash, and 127 (52.9%) were initially suspended as the result of MAB review. The following factors were significantly related to retiring from driving, initial MAB suspension and greater age. Of the 127 drivers who were initially suspended, 82 (64.6%) retired from driving, and 45 (35.4%) pursued further licensure (p <0.01). In contrast, the percentage of non-suspended drivers who did or did not pursue further licensure was similar; being 48.7% vs 51.3%. Among drivers ≥85 years of age, 68.9% retired from driving, compared with 50% of the drivers who were 75 to 84 years of age (p <0.01) While not statisically significant, higher percentages of driving retirement were noted for the following: sex - a greater percentage of men compared to women (61.9% vs 49.5%); confusion at the traffic scene (confused, 57.1% vs non-confused, 42.9%); and crash involvement (56.7% who crashed, retired vs 43.3% of those who did not crash, retired). Overall, the most important finding of this study is that as a result of police referral, only one-fifth (20.4%) of senior drivers 75 years of age or older, continued to maintain their driving privilege. However, only 40 drivers (16.7%) retained their original driving privilege without added restrictions. The data suggest that senior drivers who are not medically fit to drive may be identified by police referrals to a licensing agency. Driving occupational therapy assessments and training, and additional driving restrictions are recommended to facilitate continuation of the driving privilege for some drivers.

Zebrafish fat-free, a novel Arf effector, regulates phospholipase D to mediate lipid and glucose metabolism.

Zebrafish fat-free mutants (ffr) exhibit defective intestinal lipid metabolism and fat-free protein (Ffr) is involved in Golgi-related vesicular trafficking. In this study, we show that ffr mutants also display defective glucose metabolism. Using microarray and real-time PCR, we found that a ffr mutant with a nonsense mutation exhibits increased transcript level of ADP-ribosylation factor gene (arfs). Further analysis indicated that Ffr contains a putative Arf binding motif and can bind GTP-bound Arfs. In addition, ffr exhibited increased transcript and activity levels of the Arf downstream effector phospholipase D (PLD). Inhibition of PLD partially restored lipid and glucose metabolism in ffr, suggesting that Ffr is involved in a pathway regulating PLD activity by regulating Arfs. We propose that local over-production of phosphatidic acid (PA) by excess PLD promotes membrane curvature, which affects Golgi membrane structure and secretory processes, contributing to impairment of lipid and glucose metabolism.

Zebrafish type XVII collagen: gene structures, expression profiles, and morpholino "knock-down" phenotypes.

The human COL17A1 gene encodes type XVII collagen (also known as the 180-kDa bullous pemphigoid antigen), an integral component of hemidesmosomes, attachment complexes providing integrity to the dermal-epidermal junction. Zebrafish, a useful model system to study skin development, displays fully developed hemidesmosomes at approximately 5 days post-fertilization (dpf). We have identified two COL17A1 orthologues in the zebrafish genome, col17a1a and col17a1b, which are expressed in the skin and the neural system, respectively. The proteins coded by these genes have structural module organizations homologous to the human type XVII collagen. "Knock-down" of the expression of col17a1a with a specific morpholino targeting the 5' UTR of the gene resulted in a blistering phenotype and in perturbations in the basement membrane zone. "Knock-down" of col17a1b expression resulted in ablation or in marked reduction of neuromasts in the lateral line. Thus, zebrafish has two COL17A1 orthologues which may have evolved tissue-specific functions during vertebrate development. Collectively, zebrafish provides a model system to study the molecular aspects of skin development and offers insights into the corresponding human diseases.

The abcc6a gene expression is required for normal zebrafish development.

Pseudoxanthoma elasticum (PXE) is caused by mutations in the ABCC6 gene, which encodes a putative efflux transporter, ABCC6. The zebrafish (Danio rerio) has two ABCC6-related sequences. To study the function of abcc6 during zebrafish development, the mRNA expression levels were measured using RT-PCR and in situ hybridization. The abcc6a showed a relatively high level of expression at 5 days post-fertilization (d.p.f.) and the expression was specific to the Kupffer's vesicles. The abcc6b expression was evident at 6 hours post-fertilization (h.p.f.) and remained high up to 8 d.p.f., corresponding to embryonic kidney proximal tubules. Morpholinos were designed to both genes to prevent pre-mRNA splicing and block translation. Injection of the abcc6a morpholinos into 1-4 cell zebrafish embryos decreased gene expression by 54-81%, and induced a phenotype, pericardial edema and curled tail associated with death at around 8 d.p.f. Microinjecting zebrafish embryos with full-length mouse Abcc6 mRNA together with the morpholino completely rescued this phenotype. No phenotypic changes were observed when the abcc6b gene morpholino was injected into embryos with knock-down efficiency of 100%. These results suggest that abcc6a is an essential gene for normal zebrafish development and provide insight into the function of ABCC6, the gene mutated in PXE.

Gene expression profiles for predicting the efficacy of the anticancer drug 5-fluorouracil in breast cancer.

Chemotherapy is an important postsurgery adjuvant therapy in the treatment of breast cancer. However, because of the individual genotype differences of patients, the drug efficacy differs from person to person, even when the same chemotherapy drug is administered. The purpose of this research was to probe the gene expression profiles to predict the efficacy of 5-fluorouracil (5-FU), the common drug used in chemotherapy for various type of cancers, in Taiwanese breast cancer patients. Microarray analysis was conducted on the cancer cell line ZR-75-1 with and without 5-FU stimulation to identify the differentially expressed genes. The significant overexpressed gene groups were selected after bioinformatics software analysis to explore the molecular mechanism of 5-FU. Six strains of breast cancer cell line purchased from American Type Culture Collection were used to analyze the expression profiles of the above target gene groups. IL18, CCL28, CXCL2, SOD1, HRAS, FDXR, and CHI3L1 genes were significantly differentially expressed in 5-FU responder and nonresponder cell lines. The selected gene groups were validated with 20 strains of breast cancer primary cultures established previously in our laboratory. The experimental results demonstrated that FAM46A, IL18, CCL28, TNF, CXCL2, PLEKHA8, HRAS, FDXR, and CHI3L1 genes showed statistically significant differential expression between primary breast cancer culture cells that respond and nonrespond to 5-FU. Six genes, IL18, CCL28, CXCL2, HRAS, FDXR, and CHI3L1, showed significant differential expression pattern in both American Type Culture Collection and primary breast cancer cultured cells. The findings of this study may serve as basis for predicting the effectiveness of 5-FU on breast cancer.

Crash test ratings and real-world frontal crash outcomes: a CIREN study.

OBJECTIVE: To establish whether the Insurance Institute for Highway Safety (IIHS) offset crash test ratings are linked to different mortality rates in real world frontal crashes. METHODS: The study used Crash Injury Research Engineering Network drivers of age older than 15 years who were involved in frontal crashes. The Crash Injury Research Engineering Network is a convenience sample of persons injured in crashes with at least one Abbreviated Injury Scale score of 3+ injury or two Abbreviated Injury Scale score of 2+ injuries who were either treated at a Level I trauma center or died. Cases were grouped by IIHS crash test ratings (i.e., good, acceptable, marginal, poor, and not rated). Those rated marginal were excluded because of their small numbers. Mortality rates experienced by these ratings-based groups were compared using the Mantel-Haenszel chi test. Multiple logistic regression models were built to adjust for confounders (i.e., occupant, vehicular, and crash factors). RESULTS: A total of 1,226 cases were distributed within not rated (59%), poor (12%), average (16%), and good (14%) categories. Those rated good and average experienced a lower unadjusted mortality rate. After adjustment by confounders, those in vehicles rated good experienced a lower risk of death (adjusted OR 0.38 [0.16-0.90]) than those in vehicles rated poor. There was no significant effect for "acceptable" rating. Other factors influencing the occurrence of death were age, DeltaV >or=70 km/h, high body mass index, and lack of restraint use. CONCLUSION: After adjusting for occupant, vehicular, and crash factors, drivers of vehicles rated good by the IIHS experienced a lower risk of death in frontal crashes.

Vehicle model year and crash outcomes: a CIREN study.

OBJECTIVE: To quantify the effect of model year (MY) on the occurrence of severe injuries and death after involvement in motor vehicle crashes. METHODS: Cases involving adult front seat occupants of vehicles MY > or = 1994 equipped with frontal airbags were selected from the Crash Injury Research and Engineering Network (CIREN) database. Cases were grouped by MY: 1994-1997, 1998-2004, and 2005-2007 (MY groups [MYG] 1, 2, and 3, respectively. MYGs were compared in relation to mortality, Injury Severity Score (ISS), and the occurrence of Abbreviated Injury Severity score (AIS) 3+ and AIS4+ injuries to each body region using Mantel Haenszel chi-square, Kruskal-Wallis, and Bonferroni corrected t test. To adjust for confounders, multiple logistic regression models were built to explore the association of MYG with death. Covariates included age, BMI, delta v, principal direction of force (PDOF), restraint use, and vehicle type. RESULTS: A total of 1888 cases was distributed within MYG1 (34%), MYG2 (62%), and MYG (34%). Age, gender, BMI, and PDOF distribution did not differ among MYGs. Though ISS distribution was not different, a decrease in the occurrence of AIS4+ of the thorax and spine regions was noted over time. Mortality also decreased over time (18, 9, and 4% MYG 1, 2, and 3, respectively). Multivariate analysis revealed a protective effect of MYG2 and MGY3 (odds ratio [OR], 0.57 [0.44-0.75] and 0.22 [0.07-0.50], respectively) in relation to death. CONCLUSIONS: Front seat occupants of later MY vehicles injured during crashes experience a decreased likelihood of very severe thoracic injuries, spinal injuries, and death.

Crash and occupant predictors of pulmonary contusion.

OBJECTIVES: Pulmonary contusions (PCs) are a common injury sustained in motor vehicle collisions. The crash and occupant characteristics of PC in motor vehicle collisions are currently unknown. Additionally, the clinical significance and the impact on mortality have not been determined. METHODS: A retrospective review of the Crash Injury Research and Engineering Network database with inclusion criteria of frontal (F) and near-side lateral (L) crashes involving occupants older than 15 years, yielded 2,184 case occupants. Pearson's chi and multivariate logistic regression were used with a p < 0.05 conferring statistical significance. RESULTS: Median age was 38 years, 80% were drivers and mortality was 16%. Forty-nine percent of case occupants were not wearing lap-shoulder belts. Chest trauma was sustained by 1,131 (52%), of whom 379 had PC. Crash characteristics included: 38 kph median change in velocity (delta V), 72% frontal deformation, and 35% struck a fixed object. Injury characteristics included median Injury Severity Score 17 with the following Abbreviated Injury Score (AIS) >2 injuries: thoracic 40%, abdominal 19%, and head 24%. Univariate predictors of PC included: age <25, male, higher Injury Severity Score, fatality, delta V >45 kph, L impacts, and collision with fixed object. PC was significantly associated with occupant compartment intrusion in F but not L crashes. In multivariate analysis, significant predictors of PC included: age <25 (odds ratios [OR] = 1.5), delta V >45 kph (OR = 1.9), and fixed object (frontal crash only) (OR = 1.8). Controlling for head, spine, abdominal, and extremity injuries AIS >2, PC was not a statistically significant risk factor for mortality. This was consistent whether or not another AIS >2 thoracic injury was present. The effectiveness of side-impact airbags was not evaluated due to the small sample size. CONCLUSIONS: Crash severity as demonstrated by higher delta V was strongly associated with PC in all crashes. Frontal crashes with a fixed object or intrusion are more likely to result in a PC. The risk of PC is greatly increased in near-side lateral impacts regardless of intrusion or object struck; suggesting occupant proximity may be the most important factor. Further investigations of the efficacy of side airbags as a counter measure should be considered and continued public education of the efficacy of lap-shoulder restrains should continue. Unexpectedly, although a marker for crash severity, PC is not an independent marker of mortality.