Pulsed light and antimicrobial combination treatments for surface decontamination of cheese: Favorable and antagonistic effects.

Postprocessing cross-contamination of cheese can lead to both food safety issues and significant losses due to spoilage. Pulsed light (PL) treatment, consisting of short, high-energy, broad-spectrum light pulses, has been proven effective in reducing the microbial load on cheese surface. As PL treatment effectiveness is limited by light-cheese interactions, the possibility to improve its effectiveness by combining it with the antimicrobial nisin was explored. The effect of natamycin, which is added to cheeses as an antifungal agent, on PL effectiveness was also investigated. Pseudomonas fluorescens, Escherichia coli ATCC 25922, and Listeria innocua were used as challenge microorganisms. Bacterial cultures in stationary growth phase were diluted to initial inoculum levels of 5 or 7 log cfu per cheese slice. Slices of sharp white Cheddar cheese and white American singles were cut in rectangles of 2.5 × 5 cm. For cheese slices receiving antimicrobial treatment before PL, slices were dipped in natamycin or nisin, spot inoculated with 100 µL of bacterial suspension, and then treated with PL. Cheese slices receiving PL treatment before antimicrobials were spot inoculated, treated with PL, and then treated with antimicrobials. The PL fluence levels from 1.02 to 12.29 J/cm2 were used. Survivors were enumerated by standard plate counting or the most probable number technique, as appropriate. All treatments were performed in triplicate, and the data were analyzed using a general linear model. Treatment with nisin or natamycin before PL decreased the effectiveness of PL for all bacteria tested. For instance, PL reduced P. fluorescens on Cheddar cheese by 2.19 ± 0.27 log after 6.14 J/cm2, whereas combination treatments at the same PL fluence yielded barely 1 log reduction. Inactivation of L. innocua on Cheddar was only 0.78 ± 0.01 log when using PL after nisin, compared with a 1.30 ± 0.76 log reduction by nisin alone. This was attributed to the absorption of UV light by the 2 antimicrobials, which diminished the UV fluence received by the bacteria. Increased inactivation was obtained when antimicrobials were applied after PL. On process cheese, a maximum reduction of 3.73 ± 0.96 log of L. innocua was obtained at 9.22 J/cm2 for PL followed by nisin, compared with 3.01 ± 0.48 by PL alone. This study demonstrates that antimicrobials may increase the antimicrobial effectiveness of PL on cheese surface, but the order of treatments is critical.

Downregulation of COMMD1 by miR-205 promotes a positive feedback loop for amplifying inflammatory- and stemness-associated properties of cancer cells.

Sustained activation of nuclear factor-κB (NF-κB) in cancer cells has been shown to promote inflammation, expansion of cancer stem cell (CSC) population, and tumor development. In contrast, recent studies reveal that CSCs exhibit increased inflammation due to constitutive NF-κB activation; however, the underlying molecular mechanism remains unclear. In the present study, the analysis of microarray data revealed upregulation of NF-κB-regulated pro-inflammatory genes and downregulation of copper metabolism MURR1 domain-containing 1 (COMMD1) during the enrichment for stemness in SAS head and neck squamous-cell carcinoma (HNSCC) cells. The 3'-UTR of COMMD1 mRNA contains microRNA (miR)-205 target site. Parallel studies with HNSCC and NSCLC cells indicated that miR-205 is upregulated upon NF-κB activation and suppresses COMMD1 expression in stemness-enriched cancer cells. COMMD1 negatively regulates the inflammatory responses induced by TLR agonists, IL-1ß, and TNF-α by targeting RelA for degradation. The shRNA-mediated downregulation of COMMD1 in cancer cells enhanced inflammatory response, generating favorable conditions for macrophage recruitment. In addition, genes associated with stemness were also upregulated in these cells, which exhibited increased potential for anchorage-independent growth. Furthermore, COMMD1 downregulation promoted in vivo tumorigenesis and tumor growth, and tumors derived from COMMD1-knockdown cells displayed elevated level of NF-κB activation, increased expression of inflammatory- and stemness-associated genes, and contain expanded population of tumor-associated leukocytes and stemness-enriched cancer cells. These results suggest that COMMD1 downregulation by miR-205 promotes tumor development by modulating a positive feedback loop that amplifies inflammatory- and stemness-associated properties of cancer cells.

Pulsed-light inactivation of pathogenic and spoilage bacteria on cheese surface.

Cheese products are susceptible to postprocessing cross-contamination by bacterial surface contamination during slicing, handling, or packaging, which can lead to food safety issues and significant losses due to spoilage. This study examined the effectiveness of pulsed-light (PL) treatment on the inactivation of the spoilage microorganism Pseudomonas fluorescens, the nonenterohemorrhagic Escherichia coli ATCC 25922 (nonpathogenic surrogate of Escherichia coli O157:H7), and Listeria innocua (nonpathogenic surrogate of Listeria monocytogenes) on cheese surface. The effects of inoculum level and cheese surface topography and the presence of clear polyethylene packaging were evaluated in a full factorial experimental design. The challenge microorganisms were grown to early stationary phase and subsequently diluted to reach initial inoculum levels of either 5 or 7 log cfu/slice. White Cheddar and process cheeses were cut into 2.5×5 cm slices, which were spot-inoculated with 100 µL of bacterial suspension. Inoculated cheese samples were exposed to PL doses of 1.02 to 12.29 J/cm(2). Recovered survivors were enumerated by standard plate counting or the most probable number technique, as appropriate. The PL treatments were performed in triplicate and data were analyzed using a general linear model. Listeria innocua was the least sensitive to PL treatment, with a maximum inactivation level of 3.37±0.2 log, followed by P. fluorescens, with a maximum inactivation of 3.74±0.8 log. Escherichia coli was the most sensitive to PL, with a maximum reduction of 5.41±0.1 log. All PL inactivation curves were nonlinear, and inactivation reached a plateau after 3 pulses (3.07 J/cm(2)). The PL treatments through UV-transparent packaging and without packaging consistently resulted in similar inactivation levels. This study demonstrates that PL has strong potential for decontamination of the cheese surface.

Cerebral hemodynamics and baroreflex sensitivity after carotid artery stenting.

OBJECTIVES: The long-term hemodynamic effects of carotid angioplasty and stenting (CAS) are unclear. We performed a longitudinal study to investigate the variations in cerebral hemodynamics in patients undergoing CAS. MATERIALS AND METHODS: We performed prospective evaluation of 63 symptomatic male patients (19 patients had transient ischemic attack and 44 had minor stroke; mean age: 77.3 ± 6.3 years [range: 51-86]). The mean blood flow velocities (MBFV) and pulsatility index (PI) of the middle cerebral arteries (MCA) on both sides were evaluated using transcranial color-coded Doppler (TCCD) ultrasonography. Cardiac autonomic activities were evaluated by measuring baroreflex sensitivity (BRS). All parameters were measured at baseline prior to CAS and at 1, 3, 6, and 12 months after CAS. RESULTS: The preoperative MBFV and PI of the ipsilateral MCA were significantly lower than those of the contralateral side. However, after CAS, MBFV in the ipsilateral MCA increased significantly until 2 weeks after stenting, after which the MBFV gradually decreased and remained stable for 1 year after CAS. Further, we observed a nonsignificant increase in MBFV in the contralateral MCA after CAS. In contrast to the MBFV, the BRS values decreased significantly 1 month after stenting and returned to baseline levels 6 months after CAS. CONCLUSIONS: Patients with CAS showed improved global cerebral hemodynamic status. However, the BRS did not normalize initially, and baseline value was achieved at 6 months after stenting.

Contribution of intracranial vertebral artery asymmetry to vestibular neuropathy.

OBJECTIVES: To test the hypothesis that vertebral artery hypoplasia (VAH) may affect the lateralisation of vestibular neuropathy (VN), probably through haemodynamic effect on the vestibular labyrinth. METHODS: 69 patients with unilateral VN were examined with a magnetic resonance angiographic (MRA) and caloric test. 50 healthy subjects served as controls. The diagnosis of intracranial VAH was based on MRA if <0.22 cm in VA diameter and a diameter asymmetry index >40%. The authors then correlated the canal paretic side with the VAH side. RESULTS: MRA study revealed 29 VAH (right/left: 23/6) in VN subjects and six VAH in controls (right/left: 5/1). The RR of VAH in VN subjects compared with controls was elevated (RR=2.2; 95% CI 1.8 to 2.8). There was a high accordance rate between the side of VAH and VN. Among 29 patients with unilateral VAH, 65.5% (N=19) had an ipsilateral VN, in which left VAH showed a higher accordance rate (83.3%) than the right side (60.9%). VN subjects with vascular risk factors also had a higher VAH accordance rate (81%) than those without (25%). CONCLUSIONS: VAH may serve as a regional haemodynamic negative contributor and impede blood supply to the ipsilateral vestibular labyrinth, contributing to the development of VN, which could be enhanced by atherosclerotic risk factors and the left-sided location.

Cr(VI) removal on fungal biomass of Neurospora crassa: the importance of dissolved organic carbons derived from the biomass to Cr(VI) reduction.

Interactions of toxic Cr(VI) with renewable biomaterials are considered an important pathway for Cr(VI) removal in ecosystems. Biomaterials are susceptible to dissolution, and their dissolved derivatives may provide an alternative to surface-involved pathway for scavenging of Cr(VI). In this study, dissolved organic carbon (DOC) derived from Neurospora crassa biomass was investigated. The proportion of Cr(VI) reduction by DOC to that on biomass was determined to evaluate the importance of DOC to Cr(VI) reduction. A rapid increase in DOC concentration from 145.6 to 193.7 mg L(-1) was observed when N. crassa-biomass was immersed in 0.01 M KCl solution at pH of 1-5, and polysaccharides, peptides, and glycoproteins with carboxyl, amide, and -NH functional groups, are the major compositions of DOC. On reaction of 96.2 microM Cr(VI) with N. crassa-biomass or DOC, it was estimated that DOC contributed approximately 53.8-59.5% of the total Cr(VI) reduction on biomass in the dark. Illumination enhanced Cr(VI) reduction via photo-oxidation of biomass/DOC under aeration conditions, which formed superoxide for Cr(VI) reduction. At pH 1, photoinduced Cr(VI) reduction by DOC proceeded more rapidly than reduction on the biomass surface. However, at pH >3, with a decrease in Cr(VI) reduction by DOC, photon-excited biomass may become an important electron source for Cr(VI) photoreduction.

Gene expression of P-glycoprotein and cytochrome P450 3A4 in peripheral blood mononuclear cells and correlation with expression in liver.

Long-term immunosuppression in renal transplant recipients generally includes calcineurin inhibitors (CNIs), which demonstrate marked interindividual diversity and a narrow therapeutic range. In the clinical setting, it is important to reach therapeutic drug concentrations to prevent allograft rejection. The same immunosuppressant dosage leads to different drug concentrations. Therefore, we investigated factors that influence the metabolism of immunosuppressant agents. The CNIs are substrates of cytochrome P450 (CYP450) and P-glycoprotein. The CYP450 3A genotype significantly influences CNI concentration. Differences in expression of these proteins may explain interindividual pharmacokinetic variations. However, it is risky and impractical to obtain specimens from the liver in renal transplant recipients. Therefore, we investigated the correlation of gene expression between peripheral blood mononuclear cells (PBMCs) and liver parenchyma. We observed that the correlation of relative P-glycoprotein gene expression between PBMCs and liver is not significant (r2=0.03; P=.65). In addition, the correlation of CYP450 3A4 gene expression between PBMCs and liver is not strong (r2=0.23; P=.42). The expression level of CYP450 3A5 is too low to be detected in the sample from PBMCs.

N-Methyl-D-aspartate receptor antagonist MK-801 attenuates morphine tolerance and associated glial fibrillary acid protein up-regulation: a proteomic approach.

BACKGROUND: It is well known that long-term morphine administration results in tolerance, which limits the clinical use of this drug in pain management. METHODS: Male Wistar rats were randomly assigned to receive one of four different infusions: morphine [15 microg/h, intrathecal (i.t.)], saline, MK-801 (5 microg/h, i.t.) plus morphine (15 microg/h, i.t.), or MK-801 (5 microg/h, i.t.) alone. RESULTS: Morphine infusion induced a maximal antinociceptive effect on day 1 and tolerance on day 3, and the maximal anti-receptive tolerance was observed on day 5. Co-infusing MK-801 with morphine attenuated morphine's anti-receptive tolerance. Two-dimensional gel electrophoretic analysis of spinal proteins revealed that eight protein spots were up-regulated in morphine-tolerant rats, and that they were significantly inhibited by MK-801 co-infusion. Among the up-regulated proteins, glial fibrillary acid protein (GFAP), a glial-specific maker, was identified by mass spectrometry. This finding was also confirmed by Western blot analysis. CONCLUSION: Using proteomic analysis, we identified eight GFAP protein spots that were up-regulated in the dorsal horn of morphine-tolerant rat spinal cords. This up-regulation was partly inhibited by N-methyl-D-aspartate receptor antagonist MK-801 co-infusion, which suggests that GFAP protein can be considered to be a pathogenesis marker of morphine tolerance.

The removal and recovery of Cr(VI) by Li/Al layered double hydroxide (LDH).

Hexavalent Cr has been identified as one of the toxic metals commonly present in industrial effluents. Among the treatment techniques developed for removing Cr(VI) from waste waters, sorption is most commonly applied, due to its simplicity and efficiency. However, few adsorbents can be recycled and reused cost-effectively. In this study, the removal and recovery of Cr(VI) from water using Li/Al LDH was investigated. The removal of Cr(VI) by Li/Al LDH was evaluated in a batch mode. The results demonstrated that Cr(VI) adsorption onto Li/Al LDH occurs by replacing the Cl(-) that originally exists in the interlayer of the adsorbent. The degree of Cr(VI) adsorption observed for Li/Al LDH was relatively high and the process occurred rapidly; however, a portion of adsorbed Cr(VI) was gradually desorbed, due to the Li de-intercalation of Li/Al LDH. Lithium de-intercalation from Li/Al LDH with interlayer Cl(-) and interlayer Cr(VI) follows the first order kinetics and has the activation energies of 76.6 and 41.5 kJ mol(-1), respectively. The properties of thermal unstability and the high adsorption capacity of Li/Al LDH may lead to the development of an innovative technique for the removal of Cr(VI) from Cr(VI)-containing wastewater. That is, Li/Al LDH may be used as an effective adsorbent for the adsorption of Cr(VI) in an ambient environment. Following the adsorptive process, the adsorbed Cr(VI) may be released, using heated water to treat the Cr(VI)-containing Li/Al LDH particles. Through this hydrothermal treatment of the used adsorbent, Cr(VI) can be recovered and the solid product (gibbsite) can be recycled for further use.

Novel diffusion-weighted magnetic resonance imaging findings in leptomeningeal carcinomatosis: a case report.

This report presents a rare case of leptomeningeal carcinomatosis initially presenting with mental impairment and rapidly progressing to coma without any history of malignancy. In addition to highlighting the diagnostic difficulties, the linear high signal intensity along the cortex on the diffusion-weighted imaging (DWI) sequence of magnetic resonance (MR) imaging was identified accidentally. High signal change in the corresponding areas was also noted on unenhanced fluid-attenuated inversion recovery (FLAIR) MR imaging, which may be a novel method of diagnosing leptomeningeal carcinomatosis, which should be studied further.

Gene amplification and overexpression of protein phosphatase 1alpha in oral squamous cell carcinoma cell lines.

Gene amplification of chromosomal band 11q13 is observed frequently in oral squamous cell carcinomas (OSCC). Several genes have been identified in the 11q13 amplicon, including FGF3, FGF4, CCND1, EMS1 and TAOS1. Some of these genes show good correlation between gene copy number and gene expression, and are thought to play a role in driving 11q13 amplification. The PPP1CA gene, which encodes the catalytic subunit of serine/threonine protein phosphatase protein phosphatase 1alpha (PP1alpha), is also located in 11q13. Protein phosphatase 1alpha, one of the isoforms of PP1, regulates critical cellular events, such as cell cycle progression, and apoptosis. We sought to explore the possibility that PPP1CA was amplified and overexpressed in OSCC cells. Indeed, some OSCC cell lines had PPP1CA gene amplification, as analysed by fluorescence in situ hybridization. We have also demonstrated that PPP1CA gene copy number is increased in 21% of the OSCC cell lines determined by quantitative microsatellite analysis. PP1alpha RNA expression determined by quantitative reverse transcription-polymerase chain reaction was significantly higher in OSCC cell lines with 11q13 amplification compared to those without 11q13 amplification (P=0.011). The difference was even more significant between cell lines with at least three copies of the PPP1CA gene and those with less than three copies of the gene (P=0.00045). Relative PP1alpha protein levels were also significantly associated with PPP1CA gene copy number (P=0.014). Furthermore, knockdown of PP1alpha and/or cyclin D1 by small interfering RNA suppressed OSCC cell growth, at least in part by modulating pRB phosphorylation, resulting in G0 growth arrest. These data suggest that like the cyclin D1 gene, CCND1, amplification and overexpression of the PP1alpha gene, PPP1CA, may be involved in OSCC tumorigenesis and/or progression.

Evolution of migraine diagnoses in adolescents: a 3-year annual survey.

We conducted a 3-year annual questionnaire survey of the International Headache Society (IHS) migraine diagnoses among 2414 adolescents aged 13. For those with migraine without aura (IHS 1.1) at the baseline survey (n = 89), 28% and 24% retained the IHS 1.1 diagnosis at the 2nd and 3rd surveys. Only 0.5% of adolescents had a persistent IHS 1.1 diagnosis for 3 years. Of those with IHS 1.1 or migrainous disorder (IHS 1.7) (n = 449), 43% and 48% retained the IHS 1.1 or IHS 1.7 diagnosis at the 2nd and 3rd surveys. Conversion between IHS 1.1 and IHS 1.7 was common. About 5.6% of the adolescents suffered from IHS 1.1 or IHS 1.7 for all 3 years. Independent predictors for persistent IHS 1.1 or IHS 1.7 diagnosis included frequent headache (>5 days/month) (relative risk (RR) = 1.8) and pulsatile headache (RR = 1.5). The diagnosis of IHS 1.1 in adolescents was quite unstable. Conversion between IHS 1.1 and IHS 1.7 was an important cause. Factoring IHS 1.7 into the spectrum of migraine diagnoses during epidemiological surveys provides a realistic impression of the disease burden in this age group.

Identification of a gamma-tubulin-binding domain in BRCA1.

We previously reported (L-C. Hsu and R. L. White, Proc. Natl. Acad. Sci. USA, 95: 12983-12988, 1998) that hypophosphorylated BRCA1 is associated with mitotic centrosomes in vivo, perhaps through its interaction with gamma-tubulin. In vitro evidence presented here indicates that full-length BRCA1 protein generated by in vitro translation interacts with gamma-tubulin. A specific domain of BRCA1 protein, BRCA1 fragment no. 3 (BF3; amino acids 504-803), is both necessary and sufficient to bind gamma-tubulin. BF3 and gamma-tubulin coimmunoprecipitated when coexpressed in cells. In addition, expression of BF3 interfered with the interaction between BRCA1 and gamma-tubulin. Stable transformants of COS-7 cells that overexpressed BF3 showed a reduced growth rate partly because of increased apoptosis. Furthermore, overexpression of BF3 in COS-7 cells results in the accumulation of mitotic cells with multiple centrosomes and abnormal spindles. Okadaic acid, an inhibitor of protein phosphatases types 1 and 2A, induces hyperphosphorylation of BRCA1, a reduction of both BRCA1 and gamma-tubulin associated with mitotic centrosomes, and an accumulation of abnormal spindle formation. Thus, attenuating the interaction between BRCA1 and gamma-tubulin, and their association with mitotic centrosomes, may induce an increase of aneuploid cell population and contribute to tumorigenesis.

Predictors of use of highly active antiretroviral therapy (HAART) among persons with AIDS in San Francisco, 1996-1999.

Highly active antiretroviral therapy (HAART) has contributed to a decrease in AIDS-related morbidity and mortality. This study used population-based AIDS surveillance data to evaluate the prevalence and predictors of HAART use among persons with AIDS in San Francisco. Use of HAART among persons living with AIDS increased from 41% in 1996 to 72% in 1999. Fourteen percent of persons diagnosed with AIDS between 1996 and 1999 initiated HAART before their AIDS diagnosis. Use of HAART before an AIDS diagnosis increased from 5% in 1996 to 26% in 1999. In the multivariable analysis, African Americans, injection drug users, and those without insurance at the time of AIDS diagnosis were less likely to use HAART before AIDS diagnosis. Delayed initiation of HAART after AIDS was more likely to occur among African Americans, injection drug users, homeless persons, those with public insurance, and those with higher CD4 counts. Although the overall prevalence of HAART use was high, disparity in use of HAART existed by race and risk group, patient's insurance status, and facility of diagnosis. Barriers in use of treatment should be identified so all persons with AIDS can benefit from improved therapies.

Heparin-coated cardiopulmonary bypass circuits: current status.

Heparin-coated circuits have been subjected to vigorous testing, both experimentally and clinically, for the past decade. When the functions of heparin are preserved on the surface, the heparinized surface plays multiple roles in attenuating the systemic inflammatory response. These include the ability to attenuate contact activation, coagulation activation, complement activation and, directly or indirectly, platelet and leukocyte activation. The heparinized surface also renders the cardiopulmonary bypass (CPB) circuits hydrophilic and protein resistant and augments lipoprotein binding. The multifunctional nature of the heparinized surface contributes to the overall biocompatibility of the surface. Clinically, heparin-coated circuits become most effective in reducing systemic inflammatory response and in improving morbidity, mortality, and other patient outcome related parameters when material-independent blood activation is controlled or minimized through a global biocompatibility strategy. Techniques involved in the global biocompatibility strategy are readily available and are being effectively and safely practiced at several centers. With the global biocompatibility strategy, outstanding and reproducible results have been routinely achieved with conventional CPB techniques. Alternative revascularization procedures should equal or surpass conventional CPB, using best clinically proven strategies with respect to patient outcome and long-term graft patency.

MR perfusion studies of brain for patients with unilateral carotid stenosis or occlusion: evaluation of maps of "time to peak" and "percentage of baseline at peak".

Maps of "time to peak" (TTP) and "percentage of baseline at peak" (PBP) were compared with maps of conventional brain perfusion parameters, namely, mean transit time (MTT) and relative cerebral blood volume (rCBV). We performed MR perfusion studies in 11 patients. All of them had occlusion or high-grade stenosis of the unilateral carotid artery. Three areas of old infarct, 4 areas of new infarct, and 10 areas of brain without infarct were evaluated specifically. In all these cases, the TTP maps appeared similar to the MTT maps. They showed increases, normal values, or decreases at the same time in all areas evaluated. Most areas of abnormally decreased CBV had increased signal in PBP maps. In conclusion, the TTP map provided the same qualitative information as MTT. PBP seemed correlated inversely to CBV and was less sensitive in demonstrating abnormality.

Impact of protease inhibitors and other antiretroviral treatments on acquired immunodeficiency syndrome survival in San Francisco, California, 1987-1996.

The authors assessed temporal trends in acquired immunodeficiency syndrome (AIDS) survival for 15,271 persons in San Francisco, California, diagnosed between 1987 and 1996 with an opportunistic illness included in the 1987 AIDS case definition. Predictors of survival were evaluated for 5,686 persons who were diagnosed between 1993 and 1996 and met the 1993 AIDS case definition. Median survival was 19 months for persons diagnosed between 1987 and 1989, 17 months for persons diagnosed between 1990 and 1992, 15 months for persons diagnosed between 1993 and 1994, and 31 months for persons diagnosed between 1995 and 1996. Decreased mortality was associated with use of antiretroviral therapy without protease inhibitors before AIDS (relative hazard (RH) = 0.88, 95% confidence interval (CI): 0.8, 1.0) and after AIDS (RH = 0.83, 95% CI: 0.7, 0.9) and use of antiretroviral agents with protease inhibitors before AIDS (RH = 0.25, 95% CI: 0.2, 0.3) and after AIDS (RH = 0.36, 95% CI: 0.3, 0.4). Increased mortality was found for persons aged > or = 40 years (RH = 1.43, 95% CI: 1.3, 1.6), persons initially diagnosed with an opportunistic illness (RH = 1.97, 95% CI: 1.8, 2.2), and homosexual injection drug users (RH = 1.33, 95% CI: 1.2, 1.5). Survival after AIDS has increased. Treatment with antiretroviral agents, particularly protease inhibitors, strongly predicts improved survival.

Mouse type-2 retinaldehyde dehydrogenase (RALDH2): genomic organization, tissue-dependent expression, chromosome assignment and comparison to other types.

Retinaldehyde dehydrogenase (RALDH) isozymes catalyze the formation of an essential developmental modulator, retinoic acid. We determined the structural organization of mouse type-2 Raldh2 by isolation of overlapping genomic DNA clones from a phage library. The gene consists of 14 exons spanning more than 70 kb of genomic DNA. It was localized to mouse chromosome 6. Northern blot analysis revealed testis-specific expression. The RALDH genes belong to the aldehyde dehydrogenase (ALDH) multi-gene family. Three types of RALDH genes (e.g. human ALDH1/mouse Ahd2/rat RalDH(I), human ALDH11/mouse Raldh2/rat RalDH(II) and human ALDH6) are highly conserved during evolution, sharing about 70% identity at the amino acid level between any two gene types and 90% identity between any two mammalian genes of the same type. Different RALDH types show specific tissue and developmental expression patterns, suggesting (i) a regulatory mechanism of retinoic acid synthesis via different promoters of RALDH genes, and (ii) distinctive biological roles of different isozymes in embryogenesis and organogenesis.

Effect of heparin reversal and fresh platelet transfusion on platelet emboli post-cardiopulmonary bypass in a pig model.

Heparin reversal by protamine and fresh platelet transfusion may decrease bleeding complications post-cardiopulmonary bypass (CPB) and may increase the level of organ trapped platelet emboli. Platelet emboli were quantified in two groups of 12 Yorkshire pigs (30-35 kg), where 111indium labeled autologous platelets (INPLT: 850-1,200 microCi) were injected intravenously before and after CPB (BCPB, ACPB), and the platelet emboli level in intact organs and their samples (brain, heart, kidneys, lung, liver, and spleen) was quantified with an ion chamber and a gamma counter, respectively. All pigs were systemically heparinized (ACT > 400 sec). CPB was carried out at 2.5-3.5 L/min at 28 degrees C using a centrifugal pump, an oxygenator (OX:Bentley Univox 1.8 m2), an arterial filter (AF:0.25 m2), and a cardiotomy reservoir (CR: BMR 250) for 90 min. Heparin was reversed with an equivalent dose of protamine. The percent of INPLT dose (ID%, mean +/- SD) in organs of BCPB and ACPB pigs was calculated. The sequence of platelet emboli on a unit weight basis (ID%/g) had the following order: Spleen > Liver > Lung > Kidneys > Heart > Brain. The presence of significantly higher levels of emboli in brain, heart, and kidneys in the ACPB than the BCPB group suggest that platelet transfusion after heparin reversal with protamine may increase the risk of platelet emboli. However, it is an acceptable risk for patients having bleeding complications post-CPB.