Differential intracellular processing of the anthracycline drug ME2303 in doxorubicin-sensitive (A2780) and -resistant (A2780AD) human ovarian cancer cells as studied with confocal laser scanning microscopy and image analysis.
Laser scanning confocal microscopy has been used to follow the uptake and efflux of the 2-fluoroglycoside of doxorubicin, ME2303, in live cultures of the human ovarian cancer cell line A2780 and its doxorubicin-resistant variant A2780AD. Our methods combine confocal laser scanning microscopy and image analysis to examine the dynamics of anthracycline drugs in cancer cells. Cytotoxicity determined by MTT dye reduction showed that A2780AD cells were more than 400 times less sensitive to doxorubicin compared to A2780 cells but almost 9 times more sensitive to ME2303 compared to doxorubicin. The naturally fluorescent drug was tracked within live cells at 37 degrees C to provide time-course information in relation to nuclear and Golgi-associated cellular domains, as indicated by BODIPY FL ceramide-associated fluorescence. In both cell types, ME2303 was characterised by strong nuclear membrane and perinucleolar fluorescence and as a localised punctate pattern within the nucleus. A2780AD cells accumulated ME2303 in their nuclei at a much reduced rate compared to the doxorubicin-sensitive cells, and ME2303 efflux from resistant cell nuclei was approximately twice as fast as from A2780 cells. The relative uptake of ME2303 into Golgi-associated domains and the nucleus were monitored simultaneously during the initial 35 min of exposure to 10 microM ME2303 and during the first 45 min of a "chase" culture following exposure to 20 microM ME2303. ME2303 was detectable within the Golgi-associated domains of A2780 cells several minutes later and in less relative concentration than in A2780AD cells 15 min into a chase culture. Our results suggest the direct involvement of differences in drug processing via the Golgi apparatus in the expression of P-glycoprotein-related drug resistance.


Secretory expression and efficient purification of recombinant anthrax toxin lethal factor with full biological activity in E. coli.
Lethal factor (LF), a virulence factor of Bacillus anthracis, plays key roles in anthrax pathogenesis and host-pathogen interactions. The detailed mechanisms by which LF contributes to infection are still under investigation. While these studies require pure, homogeneous and reliable LF preparations, most methods reported for production of recombinant LF (rLF) in B. anthracis or Escherichia coli either are complicated or add extra residues to the protein. In this work, we modified our previous method by codon optimization and chromatograph workflow refinement and developed an improved strategy for efficient production of rLF from the periplasm of E. coli. We were able to obtain fully functional rLF with a purity above 95% and with a considerable yield of 5 mg/L. The preparation was characterized by SDS-PAGE, Western blot, and N-terminal sequencing, and the activity was validated by intoxication of macrophages and Fischer 344 rats. Our final product is suitable for most research involving drug development and mechanism analysis of anthrax pathogenesis.


Report on malignancies in the German juvenile idiopathic arthritis registry.
TNF inhibitors have markedly improved the therapeutic options for JIA patients. The report on malignancies observed in children exposed to TNF inhibitors flagged up questions about a potential increased risk especially for lymphoma. Cases with malignancies observed in the German registry have been reviewed to give detailed information on the patients' pretreatment and outcome. From 2001 to 2009, five cases of malignancies were documented in the German JIA biologics registry actually covering 1260 patients, one of each non-Hodgkin's lymphoma, Hodgkin's lymphoma, thyroid carcinoma, yolk sac carcinoma and cervical dysplasia. All patients have been exposed to a number of cytotoxic drugs including MTX, LEF, AZA, CSA before institution of TNF-α blockers. The malignant diagnosis has been given after exposure to etanercept for 3 weeks up to >6 years. Two patients have also been exposed to adalimumab or infliximab. At occurrence of the malignancy, three patients were still on etanercept, four on MTX and one on infliximab. In three patients, the malignancy occurred at adulthood. All patients recovered. While in some of the patients exposed to TNF inhibitors the exposure time was too short for a meaningful relationship, in others the extensive pretreatment may also be causative. Two lymphomas have been observed while other entities occurred only once. JIA patients exposed to biologics or cytotoxic drugs have to be followed carefully and long-term observation has to be continued in adulthood.


High-resolution melting curve analysis of genomic and whole-genome amplified DNA.
High-resolution melting curve analysis is an accurate method for mutation detection in genomic DNA. Few studies have compared the performance of high-resolution DNA melting curve analysis (HRM) in genomic and whole-genome amplified (WGA) DNA. In 39 paired genomic and WGA samples, 23 amplicons from 9 genes were PCR amplified and analyzed by high-resolution melting curve analysis using the 96-well LightScanner (Idaho Technology). We used genotyping and bidirectional resequencing to verify melting curve results. Melting patterns were concordant between the genomic and WGA samples in 823 of 863 (95%) analyzed sample pairs. Of the discordant patterns, there was an overrepresentation of alternate melting curve patterns in the WGA samples, suggesting the presence of a mutation (false positives). Targeted resequencing in 135 genomic and 136 WGA samples revealed 43 single nucleotide polymorphisms (SNPs). All SNPs detected in genomic samples were also detected in WGA. Additional genotyping and sequencing allowed the classification of 628 genomic and 614 WGA amplicon samples. Heterozygous variants were identified by non-wild-type melting pattern in 98% of genomic and 97% of WGA samples (P = 0.11). Wild types were correctly classified in 99% of genomic and 91% of WGA samples (P < 0.001). In WGA DNA, high-resolution DNA melting curve analysis is a sensitive tool for SNP discovery through detection of heterozygote variants; however, it may misclassify a greater number of wild-type samples.


Which patients are most likely to benefit from total joint arthroplasty?
To evaluate patient predictors of good outcome following total joint arthroplasty (TJA). A population cohort with hip/knee arthritis (osteoarthritis [OA] or inflammatory arthritis) ages ≥55 years was recruited between 1996 and 1998 (baseline) and assessed annually for demographics, troublesome joints, health status, and overall hip/knee arthritis severity using the Western Ontario and McMaster Universities OA Index (WOMAC). Survey data were linked with administrative databases to identify primary TJAs. Good outcome was defined as an improvement in WOMAC summary score greater than or equal to the minimal important difference (MID; 0.5 SD of the mean change). Logistic regression and Akaike's information criterion were used to determine the optimal number of predictors and the best model of that size. Log Poisson regression was used to determine the relative risk (RR) for a good outcome. Primary TJA was performed in 202 patients (mean age 71.0 years; 79.7% female; 82.7% with >1 troublesome hip/knee; 65.8% knee replacements). Mean improvement in WOMAC summary score was 10.2 points (SD 18.05; MID 9 points). Of these patients, 53.5% experienced a good outcome. Four predictors were optimal. The best 4-variable model included pre-TJA WOMAC, comorbidity, number of troublesome hips/knees, and arthritis type (C statistic 0.80). The probability of a good outcome was greater with worse (higher) pre-TJA WOMAC summary scores (adjusted RR 1.32 per 10-point increase; P < 0.0001), fewer troublesome hips/knees (adjusted RR 0.82 per joint; P = 0.002), OA (adjusted RR for rheumatoid arthritis versus OA 0.33; P = 0.009), and fewer comorbidities (adjusted RR per condition 0.88; P = 0.01). In an OA cohort with a high prevalence of multiple troublesome joints and comorbidity, only half achieved a good TJA outcome, defined as improved pain and disability. A more comprehensive assessment of the benefits and risks of TJA is warranted.


Contrast-Enhanced Ultrasound for Assessing Renal Perfusion Impairment and Predicting Acute Kidney Injury to Chronic Kidney Disease Progression.
Acute kidney injury (AKI) is increasingly recognized as a major risk factor leading to progression to chronic kidney disease (CKD). However, the diagnostic tools for predicting AKI to CKD progression are particularly lacking. Here, we tested the utility of contrast-enhanced ultrasound (CEUS) for predicting progression to CKD after AKI by using both mild (20-min) and severe (45-min) bilateral renal ischemia-reperfusion injury mice. Renal perfusion measured by CEUS reduced to 25% ± 7% and 14% ± 6% of the pre-ischemic levels in mild and severe AKI 1 h after ischemia (p < 0.05). Renal perfusion returned to pre-ischemic levels 1 day after mild AKI followed by restoration of kidney function. However, severe AKI caused persistent renal perfusion impairment (60% ± 9% of baseline levels) accompanied by progressive renal fibrosis and sustained decrease in renal function. Renal perfusion at days 1-21 significantly correlated with tubulointerstitial fibrosis 42 days after AKI. For predicting renal fibrosis at day 42, the area under the receiver operating characteristics curve of renal perfusion impairment at day 1 was 0.84. Similar changes in the renal image of CEUS were observed in patients with AKI-CKD progression. This study demonstrates that CEUS enables dynamic and noninvasive detection of renal perfusion impairment after ischemic AKI and the perfusion abnormalities shown by CEUS can early predict the progression to CKD after AKI. These results indicate that CEUS enables the evaluation of renal perfusion impairment associated with CKD after ischemic AKI and may serve as a noninvasive technique for assessing AKI-CKD progression. Antioxid. Redox Signal. 27, 1397-1411.


CD59 polymorphisms are associated with gene expression and different sexual susceptibility to pemphigus foliaceus.
Pemphigus foliaceus (PF) is an autoimmune disease, endemic in Brazilian rural areas, characterized by acantholysis and accompanied by complement activation, with generalized or localized distribution of painful epidermal blisters. CD59 is an essential complement regulator, inhibiting formation of the membrane attack complex, and mediating signal transduction and activation of T lymphocytes. CD59 has different transcripts by alternative splicing, of which only two are widely expressed, suggesting the presence of regulatory sites in their noncoding regions. To date, there is no association study with polymorphisms in CD59 noncoding regions and susceptibility to autoimmune diseases. In this study, we aimed to evaluate if CD59 polymorphisms have a possible regulatory effect on gene expression and susceptibility to PF. Six noncoding polymorphisms were haplotyped in 157 patients and 215 controls by sequence-specific PCR, and CD59 mRNA levels were measured in 82 subjects, by qPCR. The rs861256-allele-G (rs861256*G) was associated with increased mRNA expression (p = .0113) and PF susceptibility in women (OR = 4.11, p = .0001), which were also more prone to develop generalized lesions (OR = 4.3, p = .009) and to resist disease remission (OR = 3.69, p = .045). Associations were also observed for rs831625*G (OR = 3.1, p = .007) and rs704697*A (OR = 3.4, p = .006) in Euro-Brazilian women, and for rs704701*C (OR = 2.33, p = .037) in Afro-Brazilians. These alleles constitute the GGCCAA haplotype, which also increases PF susceptibility (OR = 4.9, p = .045) and marks higher mRNA expression (p = .0025). In conclusion, higher CD59 transcriptional levels may be related with PF susceptibility (especially in women), probably due to the effect of genetic polymorphism and to the CD59 role in T cell signal transduction.


Economic benefits of optimizing anchor therapy for rheumatoid arthritis.
The total cost of RA is substantial, particularly in patients with high levels of disability. There are considerable differences in cost between countries, driven in part by differences in the use of biologic therapies. Economic evaluations are needed to assess the extra cost of using these treatments and the benefits obtained, to ensure appropriate allocation of limited health care resources. The BeSt trial, evaluating four treatment strategies, found comparable medium-term efficacy but substantially higher costs with early biologic therapy. A systematic review of such cost-effectiveness analyses concluded that biologic therapy should be used after therapy has failed with less costly alternatives such as synthetic DMARDs and glucocorticoids. Optimizing such relatively low-cost therapy to improve outcomes may delay the need for biologic therapy, thereby saving costs. A simple model has confirmed the value of this approach. The addition of modified-release prednisone 5 mg/day to existing synthetic DMARD therapy in patients with active disease resulted in improvement in DAS-28 to below the threshold level for initiation of reimbursed biologic therapy in 28-34% of patients. On a conservative estimate (i.e. assuming no further benefits beyond the 12 weeks of the study and a 12-week wait-and-see approach to starting biologic therapy), cost savings amounted to nearly € 400 per patient. While treatment decisions should never be based only on cost considerations, prolonging disease control by optimizing the use of non-biologic treatments may bring benefits to patients and also economic benefits by delaying the need for biologic treatments.


A radioimmunoassay for insulin-like growth factor II specific for the C-peptide region.
Insulin-like growth factor II (IGF-II) is a human plasma peptide whose sequence is homologous to both insulin-like growth factor I/somatomedin C (IGF-I/SM-C) and human proinsulin in the A and B regions. However, there is no obvious homology in the C (connecting peptide) region. The synthetic 8-amino acid C-peptide segment of IGF-II (Ser-Arg-Val-Ser-Arg-Arg-Ser-Arg) was covalently linked to thyroglobulin to render it more antigenic. Antiserum against the IGF-II C-peptide was generated which had a titer of 1:2000 determined with [125I]IGF-II C-peptide. Half-maximum displacement was by 350 pg/ml IGF-II C-peptide or 80 ng/ml IGF-II. There was no displacement by IGF-I/SM-C, insulin, or a wide variety of peptides. There was also a high degree of species specificity of this antisera. Isoelectric focusing studies of immunoreactive IGF-II showed an apparent pI of 6-6.5. The mean (+/- SEM) level of IGF-II after acid chromatography of 28 normal adult males was 687.0 +/- 31.9 ng/ml. The mean of 8 acromegalics was 600.5 +/- 57.4 ng/ml, indistinguishable from normal. The IGF-II levels of 21 hypopituitary children were significantly lower (231.5 +/- 32.3 ng/ml). Thus, GH action appears to be necessary for normal levels of IGF-II, but excess GH does not cause an elevation above normal of IGF-II, unlike what is seen with IGF-I/SM-C. These structurally related IGF peptides have different control mechanisms and ultimately may play different functional roles. The availability of specific RIAs for the measurement of IGF-II will help to clarify its role in human physiology and disease states.


Biomarkers of oxidative stress and health risk assessment of heavy metal contaminated aquatic and terrestrial organisms by oil extraction industry in Ogale, Nigeria.
Ogale community in Rivers State, Nigeria is characterized by crude-oil contamination of its land resources. The present study aimed to evaluate the health risk and metal (Cd, Pb, Zn, Cr, and Ni) contamination level of the vegetable (Telfairia occidentalis), snail (Achatina achatina) and the catfish (Clarias gariepinus) collected from Ogale community. Samples collected from Elele Alimini community, a less polluted area was used as control. Oxidative damage was evaluated in tissues of snail and in the liver of catfish. The concentration of most of the tested metals in the food samples collected from the polluted sites were higher than those from the reference sites and in most cases exceeded the acceptable permissible limits. The accumulation of the metals by the food samples followed the order: T. occidentalis > A. achatina > C. gariepinus. The tissues of the snail from the polluted sites showed higher malondialdehyde (MDA) and lower glutathione (GSH) levels, and higher MDA and GSH levels in the fish liver compared to control values. The health risks associated with these metals in terms of dietary intake and target hazard quotients (THQs) showed higher non-carcinogenic effect and carcinogenic risks especially for Pb and Cd from the ingestion of Telfairia occidentalis and Achatina achatina from polluted sites. The health hazards due to metal pollution for the highly-exposed consumers of the food samples, especially in Ogale require attention. The oxidative stress response to accumulation status of metals provides a relevant tool for the assessment of metal pollution.


Possible Digenic Disease in a Caucasian Family with COL4A3 and COL4A5 Mutations.
Microscopic hematuria is a common feature of patients with Alport syndrome, a familial nephropathy due to mutations in COL4A3, COL4A4 or COL4A5. These genes encode for α3, α4, and α5 type IV collagen polypeptide chains (collagen IV α345), crucial for the structural component of the glomerular basement membrane. Even patients with mild phenotype, namely isolated microhematuria (X-linked females with thin basement membrane on electron microscopy or heterozygous carriers of COL4A3 or COL4A4 mutations), can potentially progress to proteinuria and to end-stage renal disease. Recent pedigree analyses provided evidence for digenic inheritance of Alport syndrome by concomitant mutations in COL4A3/COL4A4 or COL4A4/COL4A5. We describe a Caucasian family with concomitant COL4A3 and COL4A5 mutations, consisting of a novel c.4484A>G COL4A3 (p.Gln1495Arg) mutation and a previously reported c.1871G>A COL4A5 (p.Gly624Asp) mutation. Our segregation analysis raises the possibility that Alport syndrome resembles also digenic inheritance by COL4A3/COL4A5.


Common and specific effects of the two major forms of prolactin in the rat testis.
Prolactin (PRL) has both stimulatory and inhibitory effects on testicular function, a finding we hypothesized may be related in some part to the form of the hormone present or administered. In the analysis of the pituitary secretion profiles of early pubescent vs. mature male rats, we found PRL released from early pubescent pituitaries had about twice the degree of phosphorylation. Treatment of mature males with either unmodified PRL (U-PRL) or phosphorylated PRL (via the molecular mimic S179D PRL) for a period of 4 wk (circulating level of approximately 50 ng/ml) showed serum testosterone decreased by approximately 35% only by treatment with the phospho-mimic S179D PRL. Given the specificity of this effect, it was initially surprising that both forms of PRL decreased testicular expression of 3beta-hydroxysteroid dehydrogenase and steroidogenic acute regulatory protein. Both forms also increased expression of the luteinizing hormone receptor, but only S179D PRL increased the ratio of short to long PRL receptors. Endogenous PRL and luteinizing hormone levels were unchanged in all groups in this time frame, suggesting that effects on steroidogenic gene expression were directly on the testis. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling analysis combined with staining for 3beta-hydroxysteroid dehydrogenase and morphometric analysis showed that S179D PRL, but not U-PRL, increased apoptosis of Leydig cells, a finding supported by increased staining for Fas and Fas ligand in the testicular interstitium, providing an explanation for the specific effect on testosterone. S179D PRL, but not U-PRL, also increased apoptosis of primary spermatogonia, and U-PRL, but not S179D PRL, decreased apoptosis of elongating spermatids. Thus, in mature males, hyperprolactinemic levels of both forms of PRL have common effects on steroidogenic proteins, but specific effects on the apoptosis of Leydig and germ cells.


Physiological Functioning of Carbonic Anhydrase in the Hydrothermal Vent Tubeworm Riftia Pachyptila.
On the basis of our experiments, it is clear that carbonic anhydrase (CA) plays an important role in the CO2-concentrating mechanisms in Riftia pachyptila. Plume tissue from freshly collected animals had the highest CA activity, 253.7 +/- 36.0 {mu}mol CO2 min-1 g-1 wet wt, and trophosome activity averaged 109.4 +/- 17.9 {mu}mol CO2 min-1 g-1 wet wt. Exposure of living worms to ethoxyzolamide, a carbonic anhydrase inhibitor, resulted in a 99% decrease in CA activity (from 103.9 +/- 38.6 to 0.7 +/- 0.2 {mu}mol CO2 min-1 g-1 wet wt in the plume tissue and 57.6 +/- 17.9 to 0.04 +/- 0.11 {mu}mol CO2 min-1 g-1 wet wt in the trophosome) and essentially a complete cessation of {Sigma}CO2 uptake. High concentrations of CA appear to facilitate the equilibration between inorganic carbon (Ci) in the external and internal environments, greatly enhancing the diffusion of CO2 into the animal. In summary, R. pachyptila demonstrates very effective acquisition of inorganic carbon from the environment, thereby providing the symbionts with large amounts of CO2. This effective acquisition is made possible by three factors: extremely effective pH regulation, a large external pool of CO2, and, described in this paper, high levels of carbonic anhydrase.


Association of postmenopausal hormone replacement therapy with carotid atherosclerosis and soluble thrombomodulin: the vascular aging (EVA) study. Etude du Vieillissement Artériel.
Hormone replacement therapy (HRT) may reduce atherosclerosis among postmenopausal women, partly by reducing vascular endothelium damage. We have tested this hypothesis by evaluating the association of HRT with firstly, carotid intima media thickness (IMT) and plaques, and secondly, with endothelial cell damage, indicated by soluble thrombomodulin (sTM). Then, we tested the association between the two markers of atherosclerosis and the levels of sTM. Among 747 postmenopausal women included into the EVA study, we compared 154 HRT users (including 80% transdermal treatment) with 593 never users. Carotid IMT and plaques were measured with B-mode ultrasonography and sTM with ELISA. At least one plaque was detected among 13.6% of HRT users and 27.3% of never users. After adjustment for confounding factors, the odds ratio for the presence of plaque was 0.45 (95% confidence interval, 0.25-0.78, P=0.005) in HRT users in comparison with nonusers. HRT users had a slightly lower crude mean IMT than nonusers, but the difference was not significant. sTM was positively associated with mean IMT (P for trend=0.001) but not with plaques. Finally, estrogen users had a lower sTM level than nonusers (difference 0.14 ng/ml, P=0.03). As HRT was associated with sTM and plaques, but not with IMT, while sTM was only associated with IMT, our hypothesis was not confirmed. This suggests that the possible beneficial effects of HRT on atherosclerosis may not go through the endothelial cell damage assessed by plasma thrombomodulin.


[Dopplersonographic findings in neonates with significant persistent ductus arteriosus].
The aim of the study was the description and review of a diagnostic management for treatment of patent ductus arteriosus in preterm neonates. Indomethacin, widely used to effect nonoperative closure of patent ductus arteriosus, has been implicated in vasoactive side effects and requires an accurate diagnosis. Firstly, the hemodynamic significance of the ductus arteriosus was assessed by clinical signs, such as tachycardia, disturbed microcirculation and a high difference of central and peripheral temperature. The patent ductus arteriosus was confirmed by echocardiography. The left ventricular systolic time intervals and the cerebral perfusion were obtained by pulsed doppler recordings. 48 preterm infants below 1500 g were investigated within the first 12 hours of life and during the first week. In 32 preterm neonates (67 %) a patent ductus arteriosus without hemodynamic significance and in 9 neonates a patent ductus arteriosus with hemodynamic changes was detected. In 9 neonates there were no signs of patent ductus arteriosus. Neonates with typical clinical signs of patent ductus arteriosus exhibited significantly diminished preejection time, prolonged ejection time and a decreased quotient of preejection and ejection time. We found pathologically changed parameters of anterior cerebral artery in neonates with clinical signs of patent ductus arteriosus. To judge the efficiency of the diagnostic management the groups of neonates were compared concerning the evidence of complications. Neonates with ductus arteriosus but without therapy did not reveal more pulmonary problems as well as intracerebral hemorrhages, renal or intestinal disturbances than the group of neonates with treated ductus arteriosus. Summarizing, we suggest that the described criteria are to be taken into account before treatment of ductus arteriosus in preterm neonates. In this way a wide clinical and echocardiographical investigation will be performed in risk neonates and a useless therapy can be avoided.


Facet-dependent disorder in pristine high-voltage lithium-manganese-rich cathode material.
Defects and surface reconstructions are thought to be crucial for the long-term stability of high-voltage lithium-manganese-rich cathodes. Unfortunately, many of these defects arise only after electrochemical cycling which occurs under harsh conditions, making it difficult to fully comprehend the role they play in degrading material performance. Recently, it has been observed that defects are present even in the pristine material. This study, therefore, focuses on examining the nature of the disorder observed in pristine Li1.2Ni0.175Mn0.525Co0.1O2 (LNMCO) particles. Using atomic-resolution Z-contrast imaging and electron energy loss spectroscopy measurements, we show that there is indeed a significant amount of antisite defects present in this material, with transition metals substituting on Li metal sites. Furthermore, we find a strong segregation tendency of these types of defects toward open facets (surfaces perpendicular to the layered arrangement of atoms) rather than closed facets (surfaces parallel to the layered arrangement of atoms). First-principles calculations identify antisite defect pairs of Ni swapping with Li ions as the predominant defect in the material. Furthermore, energetically favorable swapping of Ni on the Mn sites was observed to lead to Mn depletion at open facets. Relatively, low Ni migration barriers also support the notion that Ni is the predominant cause of disorder. These insights suggest that certain facets of the LNMCO particles may be more useful for inhibiting surface reconstruction and improving the stability of these materials through careful consideration of the exposed surface.


miR-21-5p regulates mitochondrial respiration and lipid content in H9C2 cells.
Cardiovascular-related pathologies are the single leading cause of death in patients with chronic kidney disease (CKD). Previously, we found that a 5/6th nephrectomy model of CKD leads to an upregulation of miR-21-5p in the left ventricle, targeting peroxisome proliferator-activated receptor-α and altering the expression of numerous transcripts involved with fatty acid oxidation and glycolysis. In the present study, we evaluated the potential for knockdown or overexpression of miR-21-5p to regulate lipid content, lipid peroxidation, and mitochondrial respiration in H9C2 cells. Cells were transfected with anti-miR-21-5p (40 nM), pre-miR-21-5p (20 nM), or the appropriate scrambled oligonucleotide controls before lipid treatment in culture or as part of the Agilent Seahorse XF fatty acid oxidation assay. Overexpression of miR-21-5p attenuated the lipid-induced increase in cellular lipid content, whereas suppression of miR-21-5p augmented it. The abundance of malondialdehyde, a product of lipid peroxidation, was significantly increased with lipid treatment in control cells but attenuated in pre-miR-21-5p-transfected cells. This suggests that miR-21-5p reduces oxidative stress. The cellular oxygen consumption rate (OCR) was increased in both pre-miR-21-5p- and anti-miR-21-5p-transfected cells. Levels of intracellular ATP were significantly higher in anti-mR-21-5p-transfected cells. Pre-miR-21-5p blocked additional increases in OCR in response to etomoxir and palmitic acid. Conversely, anti-miR-21-5p-transfected cells exhibited reduced OCR with both etomoxir and palmitic acid, and the glycolytic capacity was concomitantly reduced. Together, these results indicate that overexpression of miR-21-5p attenuates both lipid content and lipid peroxidation in H9C2 cells. This likely occurs by reducing cellular lipid uptake and utilization, shifting cellular metabolism toward reliance on the glycolytic pathway. NEW & NOTEWORTHY Both overexpression and suppression of miR-21-5p augment basal and maximal mitochondrial respiration. Our data suggest that reliance on glycolytic and fatty acid oxidation pathways can be modulated by the abundance of miR-21-5p within the cell. miR-21-5p regulation of mitochondrial respiration can be modulated by extracellular lipids.


Imipenem in acute lung infections and bacterial re-exacerbations of chronic obstructive lung disease.
The Authors examine the changes observed in the epidemiology of lower respiratory tract infections and the role of imipenem in acute lung infections as well as in bacterial re-exacerbations of chronic obstructive lung disease. Imipenem is a new beta-lactam antibiotic characterized by an unusually broad spectrum of antibacterial activity that includes most gram-positive and gram-negative aerobic and anaerobic bacteria, by a remarkable stability to both plasmid and chromosomally-mediated beta-lactamases and by good pharmacokinetic properties. These peculiar features make this drug an attractive candidate to be used in the treatment of lower respiratory tract infections. The use of imipenem, however, should be restricted to nosocomial infections due to gram-negative aerobic resistant to other antibiotics, mixed infections due to both aerobic and anaerobic flora and as initial empirical therapy of infections in immunocompromised patients.


Epitaxial growth of a colloidal hard-sphere hcp crystal and the effects of epitaxial mismatch on crystal structure.
We demonstrate the epitaxial growth of hard-sphere hcp and double hcp crystals using a surface pattern that directly dictates the stacking sequence. A detailed three-dimensional analysis based on real-space measurements is performed on crystal structure as a function of template-crystal mismatch, which demonstrates the possibilities of colloidal epitaxy as a model system for studying the effects of a patterned substrate on crystal structure. Perfect template-induced hcp-crystal growth occurs at an isotropically deformed template. At deformed lattices we observe growth of a non-close-packed superstructure and of a perfect (100)-aligned fcc crystal. Small mismatches lead to increased out-of-plane displacements followed by a structural breakup in "crystal" grains where particle positions in successive layers are strictly periodic and "defect" grains where these positions are displaced with respect to each other. Large mismatches prevent crystallization in the surface layers. The volume fraction was found to vary drastically (up to about 20% ) as a function of template deformation.


[Pharmacokinetics of flavonoids from xiexin decoction in rats].
To study the pharmacokinetics of flavonoids from Xiexin decoction in rats. SD rats were given a single ig dose of Xiexin decoction 12 g x kg(-1), plasma and urine were collected before and after dosing. Flavonoids components in plasma and urine were measured by HPLC. Pharmacokinetic parameters were determined from the plasma concentration-time data and urinary excretion-time data with the DAS software package. Baicalin was incubated with the rat renal homogenate to investigate its metabolism in vitro. After oral administration of Xiexin decoction baicalin and wogonoside were quickly absorbed and exhibited double peak phenomena in their plasma concentrations. The first peaks in plasma concentrations of baicalin and wogonoside reached Cmax1 of (10 +/- 8) and (1.5 +/- 0.5) mg x L(-1) at Tmax1 of (0.27 +/- 0.09) and (0.17 +/- 0.00) h, while the second peaks reached Cmax2 of (3. 9 0. 5) and (0. 74 +/- 0.11) mg x L(-1) at Tmax2 of (7.6 +/- 2.6) and (16.0 +/- 0.0) h, respectively. The T(1/2) of baicalin and wogonoside were (7 +/- 3) and (6.4 +/- 2.1) h, AUC(0-infinity) were (57 +/- 12) and (15 +/- 3) mg x h x L(-1), respectively. After oral administration of Xiexin decoction, not only baicalin and wogonoside but also baicalein and wogonin can be detected in the urine. The amounts of baicalin, wogonoside, baicalein and wogonin excreted from urine during 0-72 h were (1.4 +/- 0.3), (3.4 +/- 1.3), (2.2 +/- 0.97), (10 +/- 4)% of dose given in rats, respectively. The excretion T(1/2) of the four flavonoids were (6.9 +/- 2.1), (9 +/- 4) , (8.2 +/- 2.0) and (7.2 +/- 1.8) h, respectively. Baicalin was metabolized into baicalein in the rat renal homogenate in vitro, and the kinetic parameters were measured as Vmax = 702 nmol x min(-1) x g(-1) (protein) and Km=135 micromol x L(-1). After oral administration of Xiexin decoction, flavonoids can be absorbed quickly. Only a small quantity of baicalin, wogonoside, baicalein and wogonin were excreted from urine. Baicalin may be metabolized into baicalein in the rat kidney.


[Present situation and new challenges for medical treatment of Kawasaki disease].
To predict resistance to intravenous immunoglobulin (MIG) treatment in patients with severe Kawasaki disease (KD), Gunma, Kurume, and Osaka risk scores were established. Using these scores, prospective randomised trials were performed, and addition of predni- solone or intravenous methylprednisolone -pulse to the standard regimen of IVIG improves coronary artery outcomes in severe KD. Adding the evidences of new and existing therapies, such as infliximab, cyclosporin A, ulinastatin, and plasma exchange, the clinical guideline for medical treatment of the acute phase of KD was revised in 2012. Additional large cohort studies are needed to clarify the applicability of intensive initial treatment. Challenges for the future are: 1) clarification of etiology and establishment of specific therapy, 2) establishment of personalized therapy using genetic markers related to the severity of KD.


Impact of the characteristics of patients and their clinical management on outcomes in children with homozygous familial hypercholesterolemia.
To relate clinical factors to the development of cardiovascular atherosclerosis for patients with homozygous familial hypercholesterolemia. Homozygous familial hypercholesterolemia is associated with extreme elevations in levels of cholesterol causing aggressive atherosclerosis. We reviewed 10 children, 8 of whom were male, assessed at a single institution. We found that individual characteristics, levels of lipid, cardiovascular investigations, and management were related to the activity of low density lipoprotein receptors. Activity of low density lipoprotein receptors was defined as absent, being less than 2% of normal, in 4 patients who presented at the ages of 0.3, 1.4, 1.8, and 4.5 years, respectively. The activity was minimal, representing 5%-30% of normal, in another 4 patients presenting at the ages of 6.1, 9.6, 9.9, and 12 years, and was undetermined in 2 patients who presented at the ages of 3.5, and 12.1 years. Levels of low density lipoprotein cholesterol at presentation ranged from 12.2 to 24 millimoles per litre. Plasmapheresis was performed bi-weekly in 9 patients. Patients with absence of receptor activity were less likely to have a serial decrease in the levels of low density lipoprotein cholesterol prior to plasmapheresis, and one of these patients was increased to weekly plasmapheresis. In addition, they had more aggressive cardiovascular involvement of the coronary arteries, aortic valve and aorta, requiring surgical intervention at the age of 8 and 12 years in 2 patients, with sudden death at the age of 3.1 years in one patient. In contrast, 1 patient with minimal receptor activity had surgical intervention at the age of 16.6 years and another patient died suddenly at the age of 33.6 years. Complete cardiac assessment is recommended at presentation. The frequency of plasmapheresis should be adjusted according to the activity of low density lipoprotein receptors and the individual response of the patient.


Updated Strategies and Therapies for Reducing Ischemic and Vascular Events (STRIVE) ST-segment elevation myocardial infarction critical pathway toolkit.
The Strategies and Therapies for Reducing Ischemic and Vascular Events (STRIVE) acute coronary syndromes critical pathway toolkit has been revised again based on the 2007 focused update of the American College of Cardiology/American Heart (ACC/AHA) Association guidelines for the management of patients with ST-segment elevation myocardial infarction (STEMI). A previous update of the toolkit incorporated the 2007 ACC/AHA guidelines for unstable angina/non-ST-segment elevation myocardial infarction. This review highlights the major revisions to the STEMI guidelines, and illustrates and describes the revised STRIVE critical pathway tools for STEMI, which include pathway flowcharts, standing orders, pocket cards, and posters. The updated STEMI tools are available to clinicians online on the STRIVE Website.


Do Placental Histopathologic Characteristics Differ with Gestational Ages in Preterm and Term Deliveries?
The study aim is to evaluate the placental histopathological characteristics and maternal risk factors in preterm and term births according to their weeks of gestation. We designed a prospective study involving a patient population (n = 355) composed of pregnant women who delivered preterm (n = 216) and term neonates (n = 139). The preterm births were divided into three groups as extremely (n = 22), moderate (n = 96) and late preterm (n = 98) births. The statistical analyses were performed using SPSS version 15 software. There was significant difference regarding maternal vascular underperfusion and inflammation in the extremely preterm group compared with the other groups (P = 0.001), but fetal vascular obstruction and villitis of unknown etiology were not found significantly different. According to our study results, the careful examination of the placenta of premature babies, particularly those of extremely preterm births, should be part of routine obstetrical management to determine the causes of preterm birth.


Further insight into the bioactivity of the freshwater sponge Ochridaspongia rotunda.
Bioprospection has become a dynamic scientific field that explores novel possibilities for the implementation of natural products in medicine and pharmacy. Compared to marine species from all kingdoms, freshwater species have been highly neglected. This work focuses on the screening of acetylcholinesterase inhibitory (AChE) and mutagenic activities of the acetone extract (obtained by maceration) of the freshwater sponge Ochridaspongia rotunda Arndt (Malawispongiidae) in vitro. AChE inhibitory activity was evaluated both in liquid (five different concentrations of the extract, from 1 to 100 μg/mL) and in solid (seven different concentrations of the extract, from 0.5 to 10.0 μg) by methods well described in literature, while mutagenicity was estimated using the Ames test (four different concentrations of the extract, from 0.106 to 1.328 mg/plate). Ochridaspongia rotunda acetone extract exhibited promising AChE inhibitory activity in a dose-dependent manner both in liquid (IC50 23.07 μg/mL) and in solid (1.50 μg). Furthermore, the Ames test revealed no sign of mutagenicity at any concentration tested. Its FTIR spectrum coupled with the positive Liebermann?Burchard, Salkowski and Zak color reactions (tests) indicated the presence of sterol compounds. The screened extract may inspire a search for novel anticholinesterase therapeutic agent(s) potentially used in the treatment of Alzheimer's disease. Further research will be directed toward its detailed chemical analysis along with addressing the issue of a real producer of the natural product(s) responsible for the AChE activity observed.


Sex in Penicillium series Roqueforti.
Various fungi were isolated during the course of a survey in a cold-store of apples in the Netherlands. One of these fungi belongs to the genus Penicillium and produces cleistothecia at 9 and 15 °C. A detailed study using a combination of phenotypic characters, sequences and extrolite patterns showed that these isolates belong to a new species within the series Roqueforti. The formation of cleistothecia at low temperatures and the inability to produce roquefortine C, together with a unique phylogenetic placement, make these isolates a novel entity in the Roqueforti series. The name Penicillium psychrosexualis sp. nov. (CBS 128137(T)) is proposed here for these isolates.


The methionine-rich domain of the 54 kd protein subunit of the signal recognition particle contains an RNA binding site and can be crosslinked to a signal sequence.
The 54 kd protein subunit of the signal recognition particle (SRP54) has been shown to bind signal sequences by UV crosslinking. Primary structure analysis and phylogenetic comparisons have suggested that SRP54 is composed of two domains: an amino-terminal domain that contains a putative GTP-binding site (G-domain) and a carboxy-terminal domain that contains a high abundance of methionine residues (M-domain). Partial proteolysis of SRP revealed that the two proposed domains of SRP54 indeed represent structurally discrete entities. Upon proteolysis the intact G-domain was released from SRP, whereas the M-domain remained attached to the core of the particle. Reconstitution experiments demonstrated that the isolated M-domain associates with 7SL RNA in the presence of SRP19. In addition, we observed a specific binding of the M-domain directly to 4.5S RNA of Escherichia coli, which contains a structural motif also present in 7SL RNA. This shows that the M-domain contains an RNA binding site, and suggests that SRP54 may be linked to the rest of SRP through this domain by a direct interaction with 7SL RNA. Using UV crosslinking, we found that in an in vitro translation system the preprolactin signal sequence contacts SRP through the M-domain of SRP54. These results imply that the M-domain contains the signal sequence binding site of SRP54, although we cannot exclude that the G-domain may also be in proximity to bound signal sequences.(ABSTRACT TRUNCATED AT 250 WORDS)


Survival benefit of adding chemotherapy to intensity modulated radiation in patients with locoregionally advanced nasopharyngeal carcinoma.
To evaluate the contribution of chemotherapy for patients with locoregionally advanced nasopharyngeal carcinoma (NPC) treated by intensity modulated radiotherapy (IMRT) and to identify the optimal combination treatment strategy. Between 2006 and 2010, 276 patients with stage II-IVb NPC were treated by IMRT alone or IMRT plus chemotherapy. Cisplatin-based chemotherapy included neoadjuvant or concurrent, or neoadjuvant plus concurrent protocols. The IMRT alone and chemoradiotherapy groups were well-matched for prognostic factors, except N stage, with more advanced NPC in the chemoradiotherapy arm. With a mean follow-up of 33.8 months, the 3-year actuarial rates of overall survival (OS), metastasis-free survival (MFS), relapse-free survival (RFS), and disease-free survival (DFS) were 90.3%, 84.2%, 80.3%, and 69.2% for all of the patients, respectively. Compared with the IMRT alone arm, patients treated by concurrent chemoradiotherapy had a significantly better DFS (HR = 2.64; 95% CI, 1.12-6.22; P = 0.03), patients with neoadjuvant-concurrent chemoradiotherapy had a significant improvement in RFS and DFS (HR = 4.03; 95% CI, 1.35-12.05; P = 0.01 and HR = 2.43; 95% CI, 1.09-5.44; P = 0.03), neoadjuvant chemoradiotherapy provided no significant benefit in OS, MFS, RFS, and DFS. Stage group and alcohol consumption were prognostic factors for OS and N stage was a significant predictor for DFS. Addition of concurrent or neoadjuvant-concurrent chemotherapy to IMRT is available to prolong RFS or DFS for locoregionally advanced NPC. Such work could be helpful to guide effective individualized therapy.


[Results of a serological examination of amyotrophic lateral sclerosis patients].
Examinations of specimens from 12 patients with amyotrophic lateral sclerosis (ALS) by means of enzyme-labeled immunosorbent assay (ELISA) and solid-phase radioimmunoassay ( SPIRA ) demonstrated antibody to lymphocytic choriomeningitis virus (LCM) in the blood sera of 3 patients and antigens of LCM virus in sera of 2 other patients. LCM virus antigen was also found in organs of a patient who died of ALS. LCM virus was isolated from the serum of one patient positive for LCM antigen by inoculation of newborn white mice. The new isolate (the Kanta -220 strain) examined by electron microscope was shown to have some morphological differences from the museum strain CA-1371 of LCM virus available in the laboratory.


Historical Spatial Distribution of Zoonotic Diseases in Domestic, Synanthropic, and Wild Animals in the Mexican Territory of the Yucatan Peninsula.
The Mexican territory of the Yucatan Peninsula has a tropical climate and harbors a wide variety of domestic, synanthropic, and wild animals, as well as disease vectors. To determine the distribution of recorded zoonotic diseases in the Yucatan Peninsula, scientific publications referring to these diseases in animals and containing geographic coordinates of disease occurrence, were studied. The epidemiological bulletins of the national government were also consulted to obtain information on zoonotic diseases reported in humans in the territory. The territory harbors a wide variety of tropical zoonotic pathogens, including Trypanosoma cruzi, Leptospira interrogans, Toxoplasma gondii, Leishmania mexicana, Dirofilaria immitis, and Rickettsia felis. A variety of domestic, synanthropic, and wild animals act as hosts or reservoirs in the transmission cycle of the zoonotic diseases in the Yucatan Peninsula, and some spillover into human populations has also been recorded. There are still zoonotic diseases that have rarely or never been reported in humans, but it is not clear whether this is because these diseases in humans are not common, there is a lack of viable transmission cycle or there is a lack of appropriate diagnosis. It is necessary to continue monitoring vectors, animal hosts, and humans to identify risk factors for zoonotic diseases in the Yucatan Peninsula.


The Brutal but Utile Truth of "Futile".
Though futility may be difficult to consensually define as a description of the limits of medicine, the term futility as it applies to quantitative futility is useful and apt. The term conveys a sometimes harsh but truthful reality for patients and families. When talking with patients and families, conveying the concept of futility might be more compassionately expressed as an inability to provide a curative medical benefit to the patient, with a concomitant commitment to provide comfort care. The ultimate arbiter of quantitative futility should be the medical profession.


Ganglion cell-inner plexiform layer and retinal nerve fiber layer thickness according to myopia and optic disc area: a quantitative and three-dimensional analysis.
To determine the influences of myopia and optic disc size on ganglion cell-inner plexiform layer (GCIPL) and peripapillary retinal nerve fiber layer (RNFL) thickness profiles obtained by spectral domain optical coherence tomography (OCT). One hundred and sixty-eight eyes of 168 young myopic subjects were recruited and assigned to one of three groups according to their spherical equivalent (SE) values and optic disc area. All underwent Cirrus HD-OCT imaging. The influences of myopia and optic disc size on the GCIPL and RNFL thickness profiles were evaluated by multiple comparisons and linear regression analysis. Three-dimensional surface plots of GCIPL and RNFL thickness corresponding to different combinations of myopia and optic disc size were constructed. Each of the quadrant RNFL thicknesses and their overall average were significantly thinner in high myopia compared to low myopia, except for the temporal quadrant (all Ps ≤0.003). The average and all-sectors GCIPL were significantly thinner in high myopia than in moderate- and/or low-myopia (all Ps ≤0.002). The average OCT RNFL thickness was correlated significantly with SE (0.81 μm/diopter, P < 0.001), axial length (-1.44 μm/mm, P < 0.001), and optic disc area (5.35 μm/mm2, P < 0.001) by linear regression analysis. As for the OCT GCIPL parameters, average GCIPL thickness showed a significant correlation with SE (0.84 μm/diopter, P < 0.001) and axial length (-1.65 μm/mm, P < 0.001). There was no significant correlation of average GCIPL thickness with optic disc area. Three-dimensional curves showed that larger optic discs were associated with increased average RNFL thickness and that more-myopic eyes were associated with decreased average GCIPL and RNFL thickness. Myopia can significantly affect GCIPL and RNFL thickness profiles, and optic disc size has a significant influence on RNFL thickness. The current OCT maps employed in the evaluation of glaucoma should be analyzed in consideration of refractive status and optic disc size.


DNA damage response (DDR) and senescence: shuttled inflamma-miRNAs on the stage of inflamm-aging.
A major issue in aging research is how cellular phenomena affect aging at the systemic level. Emerging evidence suggests that DNA damage response (DDR) signaling is a key mechanism linking DNA damage accumulation, cell senescence, and organism aging. DDR activation in senescent cells promotes acquisition of a proinflammatory secretory phenotype (SASP), which in turn elicits DDR and SASP activation in neighboring cells, thereby creating a proinflammatory environment extending at the local and eventually the systemic level. DDR activation is triggered by genomic lesions as well as emerging bacterial and viral metagenomes. Therefore, the buildup of cells with an activated DDR probably fuels inflamm-aging and predisposes to the development of the major age-related diseases (ARDs). Micro (mi)-RNAs - non-coding RNAs involved in gene expression modulation - are released locally and systemically by a variety of shuttles (exosomes, lipoproteins, proteins) that likely affect the efficiency of their biological effects. Here we suggest that some miRNAs, previously found to be associated with inflammation and senescence - miR-146, miR-155, and miR-21 - play a central role in the interplay among DDR, cell senescence and inflamm-aging. The identification of the functions of shuttled senescence-associated miRNAs is expected to shed light on the aging process and on how to delay ARD development.


ERRα promotes glycolytic metabolism and targets the NLRP3/caspase-1/GSDMD pathway to regulate pyroptosis in endometrial cancer.
Tumor cells can resist chemotherapy-induced pyroptosis through glycolytic reprogramming. Estrogen-related receptor alpha (ERRα) is a central regulator of cellular energy metabolism associated with poor cancer prognosis. Herein, we refine the oncogenic role of ERRα in the pyroptosis pathway and glycolytic metabolism. The interaction between ERRα and HIF-1α was verified using co-immunoprecipitation. The transcriptional binding sites of ERRα and NLRP3 were confirmed using dual-luciferase reporter assay and cleavage under targets and tagmentation (CUT&Tag). Flow cytometry, transmission electron microscopy, scanning electron microscopy, cell mito stress test, and extracellular acidification rate analysis were performed to investigate the effects of ERRα on the pyroptosis pathway and glycolytic metabolism. The results of these experiments were further confirmed in endometrial cancer (EC)-derived organoids and nude mice. In addition, the expression of ERRα-related pyroptosis genes was analyzed using The Cancer Genome Atlas and Gene Expression Omnibus database. Triggered by a hypoxic microenvironment, highly expressed ERRα could bind to the promoter of NLRP3 and inhibit caspase-1/GSDMD signaling, which reduced inflammasome activation and increased pyroptosis resistance, thereby resulting in the resistance of cancer cells to cisplatin. Moreover, ERRα activated glycolytic rate-limiting enzyme to bridge glycolytic metabolism and pyroptosis in EC. This phenomenon was further confirmed in EC-derived organoids and nude mice. CUT & Tag sequencing and The Cancer Genome Atlas database analysis showed that ERRα participated in glycolysis and programmed cell death, which resulted in EC progression. ERRα inhibits pyroptosis in an NLRP3-dependent manner and induces glycolytic metabolism, resulting in cisplatin resistance in EC cells.


Label-free detection of Alzheimer's disease through the ADP3 peptoid recognizing the serum amyloid-beta42 peptide.
The early diagnosis of Alzheimer's disease (AD) is challenging due to the lack of reliable methods for detecting its biomarkers in the noninvasive biopsies. We used surface plasmon resonance imaging to identify AD based on the detection of amyloid-beta42 in the serum by the ADP3 peptoid.


Identification of Key Functional Modules and Immunomodulatory Regulators of Hepatocellular Carcinoma.
Despite the advances in the treatment of hepatocellular carcinoma (HCC), the prognosis of HCC patients remains unsatisfactory due to postsurgical recurrence and treatment resistance. Therefore, it is important to reveal the mechanisms underlying HCC and identify potential therapeutic targets against HCC, which could facilitate the development of novel therapies. Based on 12 HCC samples and 12 paired paracancerous normal tissues, we identified differentially expressed mRNAs and lncRNAs using the "limma" package in R software. Moreover, we used the weighted gene coexpression network analysis (WGCNA) to analyze the expression data and screened hub genes. Furthermore, we performed pathway enrichment analysis based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. In addition, the relative abundance of a given gene set was estimated by single-sample Gene Set Enrichment Analysis. We identified 687 differentially expressed mRNAs and 260 differentially expressed lncRNAs. A total of 6 modules were revealed by WGCNA, and MT1M and MT1E genes from the red module were identified as hub genes. Moreover, pathway analysis revealed the top 10 enriched KEGG pathways of upregulated or downregulated genes. Additionally, we also found that CD58 might act as an immune checkpoint gene in HCC via PD1/CTLA4 pathways and regulate the levels of tumor-infiltrating immune cells in HCC tissues, which might be an immunotherapeutic target in HCC. Our research identified key functional modules and immunomodulatory regulators for HCC, which might offer novel diagnostic biomarkers and/or therapeutic targets for cancer immunotherapy.


Introduction to the special section on integrative approaches to source memory.
Source memory involves recognizing or recalling particular features that cause us to think of a mental experience as representing a particular past event. Research directed at understanding source memory comes from many domains, including cognitive-behavioral psychology, neuroscience, lifespan psychology, clinical psychology, and psychiatry. The authors believe that integrating multiple approaches to investigating source memory--most notably here, cognitive-behavioral and cognitive neuroscience--provides productive cross-fertilization of theory, methods, and analyses that in turn will yield unique constraints to our understanding of human memory. The investigations in this special section demonstrate the promise of such an integrative approach for advancing theories of source memory.


Oxygen intermediates are involved in ultraviolet radiation-induced damage of Langerhans cells.
Experiments were conducted to determine whether ultraviolet (UV) radiation exerts its effect through the generation of oxygen intermediates on Langerhans cells (LC). Guinea pigs were exposed to one single dose of UVB (0.9-2.7J/cm2), and biopsy specimens were taken 5 days after the irradiation. The population of LC was evaluated using ATPase-stained epidermal sheets. These exposures reduced the number of LC to 20-25% of the original density. On the other hand, superoxide dismutase (SOD) (0.02-0.2 mg), a scavenger of superoxide anion, which had been injected intradermally just before UV radiation, significantly prevented the depletion of LC, although not completely (37-40% of the original density). The injection immediately after the exposure was still significantly effective, but less so. Other scavengers of oxygen intermediates including catalase, D-mannitol, and L-histidine revealed no detectable effect. A single exposure of UVB at doses of 0.3-0.6 J/cm2 did not deplete the ATPase-positive LC. However, the same dose of UVB reduced the number of LC to 70%, when exposed after the injection of an SOD inactivator, diethyldithiocarbamate, possibly due to inactivation of physiologically existing SOD. These observations indicate that oxygen intermediates such as superoxide anion or its subsequent species are generated by UV radiation exposure and damage the epidermal LC.


Permanent renal sequelae secondary to drug reaction with eosinophilia and systemic symptoms (DRESS) syndrome induced by quetiapine.
We describe a Caucasian man in his late 60s who was admitted to the intensive care unit (ICU) with a history of cardiogenic shock secondary to an acute myocardial infarction. The patient's baseline serum creatinine levels were 0.9-1 mg/dL. On day 7 of the admission treatment with quetiapine was initiated due to a delirium episode. The next day the patient developed an erythematous-maculopapular rash and fever, with eosinophilia in the blood count. Over the following days the patient experienced an acute deterioration of kidney function requiring continuous renal replacement therapy. The skin lesions and eosinophilia resolved after withdrawal of quetiapine and systemic steroid therapy was administered. The patient was discharged from the ICU with a serum creatinine level of 2.6 mg/dL. Three months later, blood tests showed no recovery of the kidney function. According to the Naranjo adverse drug reaction probability scale, this event would be classified as 'probable' DRESS syndrome and, based on the RegiSCAR scoring system, was classified as 'definite' DRESS syndrome.


Selective Capture and Determination of Receptor-Binding Hemagglutinin in Influenza Vaccine Preparations Using a Coupled Receptor-Binding/RP-HPLC Assay.
Influenza vaccine potency is determined by the quantification of immunologically active hemagglutinin capable of eliciting neutralizing antibodies upon immunization. Currently, the single radial immunodiffusion (SRID) method is the standard in vitro potency assay used for lot release of seasonal inactivated influenza vaccines. Despite the proven usage of SRID, significant limitations such as the time-consuming preparation of reagents and limited dynamic range warrant the need for the development of alternative potency assays. Such alternative approaches need to discriminate and quantify relevant hemagglutinin material, provide strain identity, and be independent of strain-specific and seasonal reagents. Herein, we present a proof of concept method that combines the capture of conformationally well-folded hemagglutinin via a sialic acid binding step with the resolving power of reversed-phase high-performance liquid chromatography for strain identity and determination. Details of the protocol for the selective capture of receptor-binding hemagglutinin, its release from the receptor, and its relative determination are presented. This approach was found to provide flexibility for the reagents to be used and was adaptable to varying strain compositions of influenza vaccines. This proof of concept approach was developed as an antibody-independent methodology.


Design and synthesis of bis 1-chloromethyl-5-hydroxy-1,2- dihydro-3H-benz[e]indole (seco-CBI)-pyrrole polyamide conjugates.
[carbohydrate structure: see text] The design and synthesis of bis 1-chloromethyl-5-hydroxy-1,2-dihydro-3H-benz[e]indole (seco-CBI)-pyrrole polyamide conjugates (13, 17) as DNA minor groove binding agents are described.


C-X-C motif chemokine ligand 12: a potential therapeutic target in Duchenne muscular dystrophy.
Duchenne muscular dystrophy (DMD) is an X-linked recessive disease caused by a mutant dystrophin protein. DMD patients undergo gradual progressive paralysis until death. Chronic glucocorticoid therapy remains one of the main treatments for DMD, despite the significant side effects. However, its mechanisms of action remain largely unknown. We used bioinformatics tools to identify pathogenic genes involved in DMD and glucocorticoid target genes. Two gene expression profiles containing data from DMD patients and healthy controls (GSE38417 and GSE109178) were downloaded for further analysis. Differentially expressed genes (DEGs) between DMD patients and controls were identified using GEO2R, and glucocorticoid target genes were predicted from the Pharmacogenetics and Pharmacogenomics Knowledge Base. Surprisingly, only one gene, CXCL12 (C-X-C motif chemokine ligand 12), was both a glucocorticoid target and a DEG. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis, Gene Ontology term enrichment analysis, and gene set enrichment analysis were performed. A protein-protein interaction network was constructed and hub genes identified using the Search Tool for the Retrieval of Interacting Genes (STRING) database and Cytoscape. Enriched pathways involving the DEGs, including CXCL12, were associated with the immune response and inflammation. Levels of CXCL12 and its receptor CXCR4 (C-X-C motif chemokine receptor 4) were increased in X-linked muscular dystrophy (mdx) mice (DMD models) but became significantly reduced after prednisone treatment. Metformin also reduced the expression of CXCL12 and CXCR4 in mdx mice. In conclusion, the CXCL12-CXCR4 pathway may be a potential target for DMD therapy.


Complete Response to Pembrolizumab in Advanced Colon Cancer Harboring Somatic POLE F367S Mutation with Microsatellite Stability Status: A Case Study.
Polymerase epsilon (POLE) mutations are considered as one of the most potential and promising biomarkers for immune checkpoint inhibitors (ICIs) in patients with colorectal cancer. However, the treatment of ICIs sometimes also resulted in unsatisfactory results in patients with POLE mutations, which revealed that not all mutations on POLE contribute to tumor regression in colorectal cancer. We herein reported a case in which the patient with advanced colon cancer harboring somatic POLE F367S mutation, along with microsatellite stability status, has achieved efficacy of complete response to the programmed cell death 1 (PD-1) receptor inhibitor pembrolizumab, as well as a progression-free survival more than 49 months, and still in extension. Somatic POLE F367S mutation might be presented as a sensitive predictor to pembrolizumab in patients with colon cancer.


Legionella Pneumophila Transcriptome during Intracellular Multiplication in Human Macrophages.
Legionella pneumophila is the causative agent of Legionnaires' disease, an acute pulmonary infection. L. pneumophila is able to infect and multiply in both phagocytic protozoa, such as Acanthamoeba castellanii, and mammalian professional phagocytes. The best-known L. pneumophila virulence determinant is the Icm/Dot type IVB secretion system, which is used to translocate more than 150 effector proteins into host cells. While the transcriptional response of Legionella to the intracellular environment of A. castellanii has been investigated, much less is known about the Legionella transcriptional response inside human macrophages. In this study, the transcriptome of L. pneumophila was monitored during exponential and post-exponential phase in rich AYE broth as well as during infection of human cultured macrophages. This was accomplished with microarrays and an RNA amplification procedure called selective capture of transcribed sequences to detect small amounts of mRNA from low numbers of intracellular bacteria. Among the genes induced intracellularly are those involved in amino acid biosynthetic pathways leading to l-arginine, l-histidine, and l-proline as well as many transport systems involved in amino acid and iron uptake. Genes involved in catabolism of glycerol are also induced during intracellular growth, suggesting that glycerol could be used as a carbon source. The genes encoding the Icm/Dot system are not differentially expressed inside cells compared to control bacteria grown in rich broth, but the genes encoding several translocated effectors are strongly induced. Moreover, we used the transcriptome data to predict previously unrecognized Icm/Dot effector genes based on their expression pattern and confirmed translocation for three candidates. This study provides a comprehensive view of how L. pneumophila responds to the human macrophage intracellular environment.


Angiogenesis and lymphangiogenesis of gastric cancer.
Tumor angiogenesis is the result of an imbalance between positive and negative angiogenic factors released by tumor and host cells into the microenvironment of the neoplastic tissue. The stroma constitutes a large part of most solid tumors, and cancer-stromal cell interactions contribute functionally to tumor growth and metastasis. Activated fibroblasts and macrophages in tumor stroma play important roles in angiogenesis and tumor progression. In gastric cancer, tumor cells and stromal cells produce various angiogenic factors, including vascular endothelial growth factor, interleukin-8, platelet-derived endothelial cell growth factor, and angiopoietin. In addition, Helicobacter pylori infection increases tumor cell expression of metastasis-related genes including those encoding several angiogenic factors. We review the current understanding of molecular mechanisms involved in angiogenesis and lymphangiogenesis of human gastric cancer.


Effect of type 2 diabetes, surgical incision, and volatile anesthesia on hemodynamics in the rat.
Diabetic patients have increased cardiac complications during surgery, possibly due to impaired autonomic regulation. Anesthesia lowers blood pressure and heart rate (HR), whereas surgical intervention has opposing effects. The interaction of anesthesia and surgical intervention on hemodynamics in diabetes is unknown, despite being a potential perioperative risk factor. We aimed to determine the effect of diabetes on the integrative interaction between hemodynamics, anesthesia, and surgical incision. Zucker type 2 diabetic rats (DM) and their nondiabetic littermates (ND) were implanted with an intravenous port for drug delivery, and a radiotelemeter to measure mean arterial blood pressure (MAP) and derive HR (total n = 50). Hemodynamic pharmacological responses were assessed under conscious, isoflurane anesthesia (~2-2.5%), and anesthesia-surgical conditions; the latter performed as a laparotomy. MAP was not different between groups under conscious conditions (ND 120 ± 6 vs. DM 131 ± 4 mmHg, P > 0.05). Anesthesia reduced MAP, but not differently in DM (ND -30 ± 6 vs. DM -38 ± 4 ΔmmHg, P > 0.05). Despite adequate anesthesia, surgical incision increased MAP, which tended to be less in DM (ND +21 ± 4 vs. DM +13 ± 2 ΔmmHg, P = 0.052). Anesthesia disrupted central baroreflex HR responses to sympathetic activation (sodium nitroprusside 10 μg·kg-1, ND conscious 83 ± 13 vs. anesthetized 16 ± 5 Δbpm; P < 0.05) or to sympathetic withdrawal (phenylephrine 10 μg·kg-1, ND conscious -168 ± 37 vs. anesthetized -20 ± 6 Δbpm; P < 0.05) with no additional changes observed after surgical incision or during diabetes. During perioperative conditions, type 2 diabetes did not impact on short-term hemodynamic regulation. Anesthesia had the largest hemodynamic impact, whereas surgical effects were limited to modulation of baseline blood pressure.


The role of immune checkpoints in cardiovascular disease.
Immune checkpoint inhibitors (ICI) are monoclonal antibodies which bind to immune checkpoints (IC) and their ligands to prevent inhibition of T-cell activation by tumor cells. Currently, multiple ICI are approved targeting Cytotoxic T-lymphocyte antigen 4 (CTLA-4), Programmed Death Protein 1 (PD-1) and its ligand PD-L1, and Lymphocyte-activation gene 3 (LAG-3). This therapy has provided potent anti-tumor effects and improved prognosis for many cancer patients. However, due to systemic effects, patients can develop immune related adverse events (irAE), including possible life threatening cardiovascular irAE, like atherosclerosis, myocarditis and cardiomyopathy. Inhibition of vascular IC is associated with increased atherosclerotic burden and plaque instability. IC protect against atherosclerosis by inhibiting T-cell activity and cytokine production, promoting regulatory T-cell differentiation and inducing T-cell exhaustion. In addition, PD-L1 on endothelial cells might promote plaque stability by reducing apoptosis and increasing expression of tight junction molecules. In the heart, IC downregulate the immune response to protect against cardiac injury by reducing T-cell activity and migration. Here, inhibition of IC could induce life-threatening T-cell-mediated-myocarditis. One proposed purpose behind lymphocyte infiltration is reaction to cardiac antigens, caused by decreased self-tolerance, and thereby increased autoimmunity because of IC inhibition. In addition, there are several reports of ICI-mediated cardiomyopathy with immunoglobulin G expression on cardiomyocytes, indicating an autoimmune response. IC are mostly known due to their cardiotoxicity. However, t his review compiles current knowledge on mechanisms behind IC function in cardiovascular disease with the aim of providing an overview of possible therapeutic targets in prevention or treatment of cardiovascular irAEs.


ROCK/actin/MRTF signaling promotes the fibrogenic phenotype of fibroblast-like synoviocytes derived from the temporomandibular joint.
Malocclusion caused by abnormal jaw development or muscle overuse during mastication results in abnormal mechanical stress to the tissues surrounding the temporomandibular joint (TMJ). Excessive mechanical stress against soft and hard tissues around the TMJ is involved in the pathogenesis of inflammatory diseases, including osteoarthritis (OA). OA-related fibrosis is a possible cause of joint stiffness in OA. However, cellular and molecular mechanisms underlying fibrosis around the TMJ remain to be clarified. Here, we established a cell line of fibroblast‑like synoviocytes (FLSs) derived from the mouse TMJ. Then, we examined whether the Rho‑associated coiled‑coil forming kinase (ROCK)/actin/myocardin-related transcription factor (MRTF) gene regulatory axis positively regulates the myofibroblast (MF) differentiation status of FLSs. We found that i) FLSs extensively expressed the MF markers α‑smooth muscle actin (α‑SMA) and type I collagen; and ii) an inhibitor against the actin‑polymerizing agent ROCK, Y‑27632; iii) an actin-depolymerizing agent cytochalasin B; iv) an inhibitor of the MRTF/serum response factor‑regulated transcription, CCG‑100602, clearly suppressed the mRNA levels of α‑SMA and type I collagen in FLSs; and v) an MF differentiation attenuator fibroblast growth factor‑1 suppressed filamentous actin formation and clearly suppressed the mRNA levels of α-SMA and type I collagen in FLSs. These results strongly suggest that the ROCK/actin/MRTF axis promotes the fibrogenic activity of synoviocytes around the TMJ. Our findings partially clarify the molecular mechanisms underlying the emergence of TMJ‑OA and may aid in identifying drug targets for treating this condition at the molecular level.


Inhibition of Adenosine Kinase Attenuates Acute Lung Injury.
Extracellular adenosine has tissue-protective potential in several conditions. Adenosine levels are regulated by a close interplay between nucleoside transporters and adenosine kinase. On the basis of the evidence of the role of adenosine kinase in regulating adenosine levels during hypoxia, we evaluated the effect of adenosine kinase on lung injury. Furthermore, we tested the influence of a pharmacologic approach to blocking adenosine kinase on the extent of lung injury. Prospective experimental animal study. University-based research laboratory. In vitro cell lines, wild-type and adenosine kinase+/- mice. We tested the expression of adenosine kinase during inflammatory stimulation in vitro and in a model of lipopolysaccharide inhalation in vivo. Studies using the adenosine kinase promoter were performed in vitro. Wild-type and adenosine kinase+/- mice were subjected to lipopolysaccharide inhalation. Pharmacologic inhibition of adenosine kinase was performed in vitro, and its effect on adenosine uptake was evaluated. The pharmacologic inhibition was also performed in vivo, and the effect on lung injury was assessed. We observed the repression of adenosine kinase by proinflammatory cytokines and found a significant influence of nuclear factor kappa-light-chain-enhancer of activated B-cells on regulation of the adenosine kinase promoter. Mice with endogenous adenosine kinase repression (adenosine kinase+/-) showed reduced infiltration of leukocytes into the alveolar space, decreased total protein and myeloperoxidase levels, and lower cytokine levels in the alveolar lavage fluid. The inhibition of adenosine kinase by 5-iodotubercidin increased the extracellular adenosine levels in vitro, diminished the transmigration of neutrophils, and improved the epithelial barrier function. The inhibition of adenosine kinase in vivo showed protective properties, reducing the extent of pulmonary inflammation during lung injury. Taken together, these data show that adenosine kinase is a valuable target for reducing the inflammatory changes associated with lung injury and should be pursued as a therapeutic option.


Triple-decker-sandwich versus rice-ball structures for tris(benzene)dimetal derivatives of the first-row transition metals.
Compounds of the type M(2)Bz(3) (Bz = benzene, C(6)H(6)) have been of interest since the related triple-decker mesitylenechromium sandwich (1,3,5-Me(3)C(6)H(3))(3)Cr(2) has been synthesized and characterized structurally by X-ray crystallography. Theoretical studies predict the lowest-energy M(2)Bz(3) structures of the early transition metals Ti, V, and Cr to be the triple-decker sandwiches trans-Bz(2)M(2)(η(6),η(6)-μ-C(6)H(6)) having quintet, triplet, and singlet spin states, respectively. In these structures, the central benzene ring functions as a hexahapto ligand to each metal atom. The singlet rice-ball cis-Bz(2)M(2)(μ-C(6)H(6)) structures with a 2.64-Å Mn═Mn double bond or a 2.81-Å Fe-Fe single bond are preferred for the central transition metals Mn and Fe. Singlet triple-decker-sandwich structures trans-Bz(2)M(2)(μ-C(6)H(6)) return as the lowest-energy structures for the late transition metals Co and Ni but with the central benzene ring only partially bonded to each metal atom. Thus, the lowest-energy cobalt derivative has a trans-Bz(2)Co(2)(η(3),η(3)-μ-C(6)H(6)) structure in which the central benzene ring acts as a trihapto ligand to each metal atom. Similarly, the lowest-energy nickel derivative has a trans-Bz(2)Ni(2)(η(2),η(2)-μ-C(6)H(6)) structure in which the central benzene ring acts as a dihapto ligand to each metal atom, leaving an uncomplexed C═C double bond. The metal-metal bond orders in the singlet "rice-ball" structures cis-Bz(2)M(2)(μ-C(6)H(6)) (M = Mn, Fe) and the hapticities of the central benzene rings in the singlet late-transition-metal triple-decker-sandwich structures trans-Bz(2)M(2)(μ-C(6)H(6)) (M = Co, Ni) are governed by the desirability for the metal atoms to attain the favored 18-electron configuration.


A comparison of supervised classification methods for auditory brainstem response determination.
The ABR is commonly used in the Audiology clinic to determine and quantify hearing loss. Its interpretation is subjective, dependent upon the expertise and experience of the clinical scientist. In this study we investigated the role of machine learning for pattern classification in this domain. We extracted features from the ABRs of 85 test subjects (550 waveforms) and compared four complimentary supervised classification methods: Naïve Bayes, Support Vector Machine Multi-Layer Perceptron and KStar. The Abr dataset comprised both high level and near threshold recordings, labeled as 'response' or 'no response' by the human expert. Features were extracted from single averaged recordings to make the classification process straightforward. A best classification accuracy of 83.4% was obtained using Naïve Bayes and five relevant features extracted from time and wavelet domains. Naïve Bayes also achieved the highest specificity (86.3%). The highest sensitivity (93.1%) was obtained with Support Vector Machine-based classification models. In terms of the overall classification accuracy, four classifiers have shown the consistent, relatively high performance, indicating the relevance of selected features and the feasibility of using machine learning and statistical classification models in the analysis of ABR.


Morphological and proteomic responses of Eruca sativa exposed to silver nanoparticles or silver nitrate.
Silver nanoparticles (AgNPs) are widely used in commercial products, and there are growing concerns about their impact on the environment. Information about the molecular interaction of AgNPs with plants is lacking. To increase our understanding of the mechanisms involved in plant responses to AgNPs and to differentiate between particle specific and ionic silver effects we determined the morphological and proteomic changes induced in Eruca sativa (commonly called rocket) in response to AgNPs or AgNO3. Seedlings were treated for 5 days with different concentrations of AgNPs or AgNO3. A similar increase in root elongation was observed when seedlings were exposed to 10 mg Ag L(1) of either PVP-AgNPs or AgNO3. At this concentration we performed electron microscopy investigations and 2-dimensional electrophoresis (2DE) proteomic profiling. The low level of overlap of differentially expressed proteins indicates that AgNPs and AgNO3 cause different plant responses. Both Ag treatments cause changes in proteins involved in the redox regulation and in the sulfur metabolism. These responses could play an important role to maintain cellular homeostasis. Only the AgNP exposure cause the alteration of some proteins related to the endoplasmic reticulum and vacuole indicating these two organelles as targets of the AgNPs action. These data add further evidences that the effects of AgNPs are not simply due to the release of Ag ions.


Isolation and gene expression analysis of Arabidopsis thaliana mutants with constitutive expression of ATL2, an early elicitor-response RING-H2 zinc-finger gene.
Genes with unstable transcripts often encode proteins that play important regulatory roles. ATL2 is a member of a multigene family coding highly related RING-H2 zinc-finger proteins that may function as E3 ubiquitin ligases. ATL2 mRNA accumulation occurs rapidly and transiently after incubation with elicitors of pathogen response. We screened 50,000 M(2) families from a line that carries a fusion of pATL2 to the GUS reporter gene and isolated five mutants, which we named eca (expresión constitutiva de ATL2), that showed constitutive expression of the reporter gene. One mutant exhibits a drastic stunted phenotype while the other four grow similarly to wild type. Two early chitin-induced genes and known pathogenesis-related genes such as NPR1, PAL, and CHS are activated in all the mutants whereas members of the ATL family and PR-1 and PDF2.1, which are markers of the salicylic acid (SA) jasmonate (JA) defense-response pathways, display differential expression between the mutants. These observations indicate that the ECA gene products may function in the early steps of an elicitor-response pathway, although some of them may function at other stages on the SA or JA defense-response pathways. Likewise, the fact that ATL2 and other members of the ATL family are activated in eca mutants links the induction of this putative class of ubiquitin ligases to plant defense signaling pathways.


The type II error probability of a group sequential test of efficacy and futility, and considerations for power and sample size.
For a clinical trial incorporating a group sequential test that allows early stopping for efficacy or futility (GSTEF), the primary hypothesis concerns efficacy. However, the type II error probability of the tests of efficacy is neither specified nor known. The type II error probability of a GSTEF is partitioned into the sum of its component type II error probabilities of futility and efficacy. This partitioning provides transparency, allowing researchers flexibility to set these component error probabilities directly and to know the impact on the total type II error probability and vice versa. This transparency and flexibility should improve the application of GSTEF to clinical trials.


A Method to Assess Fc-mediated Effector Functions Induced by Influenza Hemagglutinin Specific Antibodies.
Antibodies play a crucial role in coupling the innate and adaptive immune responses against viral pathogens through their antigen binding domains and Fc-regions. Here, we describe how to measure the activation of Fc effector functions by monoclonal antibodies targeting the influenza virus hemagglutinin with the use of a genetically engineered Jurkat cell line expressing an activating type 1 Fc-FcγR. Using this method, the contribution of specific Fc-FcγR interactions conferred by immunoglobulins can be determined using an in vitro assay.


Bezoar-induced small bowel obstruction: a rare cause of a common problem.
Small Bowel Obstruction (SBO) is one of the most common diagnoses that general surgeons encounter. Adhesive disease, hernia, and neoplasm are the most common causes. A more rare cause is bezoar. A 66-year-old female with a history of prior abdominal surgery presented with clinical concern for SBO. CT scan of the abdomen and pelvis demonstrated SBO with a transition point in the left lower abdomen. The patient failed nonoperative management and was taken to the operating room for exploration. On exploration, a segment of hemorrhagic jejunum was found with an intraluminal bezoar. SBO secondary to bezoar can be managed endoscopically or operatively depending on location and size of the stone. Operative intervention can vary between laparoscopic milking of the bezoar distally, enterotomy with stone extraction, or bowel resection and anastomosis. This case illustrates the importance of maintaining a broad differential for common surgical disease processes.


Towards microbiome transplant as a therapy for periodontitis: an exploratory study of periodontitis microbial signature contrasted by oral health, caries and edentulism.
Conventional periodontal therapy aims at controlling supra- and subgingival biofilms. Although periodontal therapy was shown to improve periodontal health, it does not completely arrest the disease. Almost all subjects compliant with periodontal maintenance continue to experience progressive clinical attachment loss and a fraction of them loses teeth. An oral microbial transplant may be a new alternative for treating periodontitis (inspired by fecal transplant). First, it must be established that microbiomes of oral health and periodontitis are distinct. In that case, the health-associated microbiome could be introduced into the oral cavity of periodontitis patients. This relates to the goals of our study: (i) to assess if microbial communities of the entire oral cavity of subjects with periodontitis were different from or oral health contrasted by microbiotas of caries and edentulism patients; (ii) to test in vitro if safe concentration of sodium hypochlorite could be used for initial eradication of the original oral microbiota followed by a safe neutralization of the hypochlorite prior transplantation. Sixteen systemically healthy white adults with clinical signs of one of the following oral conditions were enrolled: periodontitis, established caries, edentulism, and oral health. Oral biofilm samples were collected from sub- and supra-gingival sites, and oral mucosae. DNA was extracted and 16S rRNA genes were amplified. Amplicons from the same patient were pooled, sequenced and quantified. Volunteer's oral plaque was treated with saline, 16 mM NaOCl and NaOCl neutralized by ascorbate buffer followed by plating on blood agar. Ordination plots of rRNA gene abundances revealed distinct groupings for the oral microbiomes of subjects with periodontitis, edentulism, or oral health. The oral microbiome in subjects with periodontitis showed the greatest diversity harboring 29 bacterial species at significantly higher abundance compared to subjects with the other assessed conditions. Healthy subjects had significantly higher abundance in 10 microbial species compared to the other conditions. NaOCl showed strong antimicrobial properties; nontoxic ascorbate was capable of neutralizing the hypochlorite. Distinct oral microbial signatures were found in subjects with periodontitis, edentulism, or oral health. This finding opens up a potential for a new therapy, whereby a health-related entire oral microbial community would be transplanted to the diseased patient.


An epidemiologic study to screen for chronic myelocytic leukemia in war victims exposed to mustard gas.
Chemical agents such as mustard gas (or sulfur mustard), which has alkylating characteristics, were used against Iranian combatants in the Iraq-Iran war. Previous studies have not shown a strong link between these chemical agents and the development of chronic myelocytic leukemia (CML). The purpose of this study was to evaluate the increased risk of CML development in Iranian soldiers exposed to mustard gas during the war. Based on a descriptive study of 2,500 cases with documented exposure to various chemical warfare agents, 665 patients had documented exposure to mustard gas. We screened the latter using the leukocyte alkaline phosphatase (LAP) test and performed further cytochemical studies on cases with positive results. From among the 665 cases with documented exposure to mustard gas, 9 cases had LAP scores < 20; 2 of these 9 cases had CML and a score of zero (0.3%). We detected cytogenetic abnormalities in 7 patients with low LAP scores and atypical lymphocytes of 5-11% in 40 patients. The risk ratio of CML developing in victims exposed to mustard gas (cutaneous or respiratory) may be higher in comparison with the normal population, although confounding factors (e.g., the possibility of exposure to combined chemical agents, excluding patients who did not manifest blisters) limited our results. Because the increased development of CML in young patients with a documented history of exposure to mustard gas cannot be disregarded, further studies are needed.


THE OCCURRENCE OF DINOFLAGELLATE LUMINESCENCE AT WOODS HOLE.
1. Bioluminescence in the water in Woods Hole was measured once a week from November, 1965, to November, 1966, and was related to the occurrence of armored luminescent dinoflagellates. 2. Thirty-two species of dinoflagellates were identified, 23 of these were tested for luminescence as isolated cells. Sixteen species were found to be luminescent. 3. Population densities of dinoflagellate species were measured by counting individuals screened from water samples. The concentrations of luminescent species varied from nearly 1000 cells per liter in February, 1966, to less than 5 cells per liter during June and July, while total dinoflagellates ranged from more than 4000 cells per liter to 5 cells per liter. 4. Amounts of luminescence radiometrically recorded in air-bubble-stimulated water samples correlated with the numbers of luminescent armored dinoflagellates present. Exceptions to this correlation were found when population densities were too high for the radiometric recording techniques to be accurate and during May and August, 1966, when the luminescent organisms were too small to be collected and counted. 5. Endogenous diurnal rhythms of flashing were found only during August, 1965 (Kelly and Katona, 1966). 6. Light inhibition of flashing was found whenever it was tested (at least once each month), although sensitivity to light inhibition varied greatly-as little as 1 second of exposure to 350 lux illumination reduced flashing by ⅔. 7. The ecology and evolution of luminescent dinoflagellates are discussed. It is suggested that dinoflagellates are responsible for most near-surface luminescence, that luminescent forms are derived from a single common ancestor, that the ability serves an adaptive function, as yet undetermined, and that much more work on a number of areas will be required before a detailed ecological understanding of dinoflagellate luminescence is obtained.


Is there a homolytic substitution chemistry (SH2) of sulfones?
[reaction: see text] A series of 2-alkylsulfonyl-2'-biphenyl radicals, in which the alkyl group is primary, secondary, or tertiary, were generated and the products of their reactions investigated. Dibenzothiophene S,S-dioxide was not identified among the products, which arose mainly from intramolecular hydrogen abstraction from the alkyl group or addition to the solvent, benzene. On this basis, it is concluded that homolytic substitution at sulfonyl sulfur, if possible at all, is too slow to take precedence over a number of competing decomposition pathways. Previous literature results suggesting the possibility of intramolecular homolytic substitution at sulfonyl sulfur may be explained by alternative processes.


Effects of a remote patient monitoring system for patients on automated peritoneal dialysis: a randomized crossover controlled trial.
Remote patient monitoring (RPM) has contributed to improved patient-centered outcomes and prognosis in patients with end-stage renal disease on automated peritoneal dialysis (APD). However, evidence from prospective trials is lacking. The participants (n = 15; median age: 65 years; males: 10; peritoneal dialysis vintage: 6.4 ± 3.5 years) randomly received APD therapy using the Kaguya® APD system either with or without the connective use of the cloud-based RPM software Sharesource® for 12 weeks. The primary outcome was patient satisfaction assessed using a modified nine-item Treatment Satisfaction Questionnaire for Medication (TSQM-9) questionnaire. The secondary outcomes were healthcare resource consumption, the health-related quality of life (HRQOL) subscales assessed with the Kidney Disease Quality of Life-Short Form questionnaire, and clinical laboratory parameters. Significant improvements were observed in the TSQM-9 subscales of Effectiveness (64.4 ± 18.8 vs. 57.8 ± 18.8; P = 0.006) and Convenience (76.3 ± 15.4 vs. 63.3 ± 17.3; P < 0.001) in patients on Sharesource®. Moreover, Sharesource® reduced the total amount of healthcare resource consumption (0.80 ± 1.32 vs. 1.87 ± 2.39 times/12 weeks; P = 0.02) and consultation time during regular monthly visits (813 ± 269 vs. 1024 ± 292 s; P < 0.001). A significant increase in ultrafiltration volume was found associated with more frequent modification of APD prescription in patients with Sharesource®. Sharesource® also improved the HRQOL subscale of General Health and Vitality. Sharesource® can improve patient-centered outcomes in patients on APD while reducing the treatment burden for both patients and medical staff. The study was registered in the Japan Registry of Clinical Trials (jRCT Number: jRCTs032190005).


Expression patterns of the native Shrunken-2 promoter in Sorghum bicolor visualised through use of the GFP reporter gene.
The AGPase large subunit (shrunken-2) promoter was demonstrated to be active in the placentochalaza and endosperm of developing grain as well as the root tips in transgenic sorghum. The temporal and spatial expression patterns of the Sorghum bicolor Shrunken-2 (Sh2) promoter were evaluated using the green fluorescence protein reporter gene (gfp) in transgenic sorghum, within the context of upregulating starch biosynthesis in the developing grain. GFP fluorescence was analysed throughout development in various tissue types using confocal laser scanning microscopy techniques. Sh2 promoter activity was first detected in the placentochalaza region of the developing caryopsis and apoplasm adjacent to the nucellar epidermis at 7 days post anthesis (dpa) where fluorescence remained relatively constant until 17 dpa. Fluorescence in this region weakened by 20 dpa and disappeared by 25 dpa. Expression was also detected in the developing endosperm, but not until 12 dpa, continuing until 25 dpa. Whilst the endosperm expression was expected, the fluorescence detected in the placentochalaza was completely unexpected. Although transcript presence does not mean the resulting biochemistry is also present, these preliminary findings may suggest alternate spatial activity of ADP-glucose pyrophosphorylase prior to uptake by the developing grain. Sh2 promoter activity was also unexpectedly detected in the root tips at all developmental time points. Sh2 promoter activity was not detected in any reproductive floral tissue (both pre and post anthesis) or in pollen. Similarly, no expression was detected in leaf tissue at any stage.


A precise and useful method for coordinating computed tomography and jaw movement frames of reference.
The aim of this study was to develop a means of coordinating helical computed tomography (CT)- based morphological data in 3 dimensions (3-D) with that pertaining to jaw movement as recorded by a device that measures jaw movement in six-degrees-of-freedom (6-DOF), thus producing multi-point movement analysis of the condyle. The study sample was two volunteers. One of the subjects had erosive bony changes in both condyles, while the other had healthy condyles. We employed a customized facebow, which enabled us to coordinate jaw movement data and morphological volume data from CT. Total uncertainty of the coordination was computed, according to International Organization for Standardization (ISO). In order to demonstrate the effects of multi-point analysis for complex condylar movement, we tried to visualize the trajectory of the working condyle in lateral excursion. The overall uncertainty at a condylar center chosen as an example to illustrate the method was 0.38 mm, 0.19 mm, and 0.50 mm in antero-posterior, lateromedial, and supero-inferior directions, respectively, in terms of 95% coverage as defined by the ISO. We developed facebow-based X-ray markers with high clinical operability, which could correlate the helical CT's coordinate system with our 6-DOF jaw movement measuring system for precise analysis of 3-D condylar movements. In motion analysis of rotational condyle, even a small amount of measurement error cannot necessarily be neglected. Then, a multi-point approach such as that realized by our system presents the best option.


Genome-wide association study of language performance in Alzheimer's disease.
Language impairment is common in prodromal stages of Alzheimer's disease (AD) and progresses over time. However, the genetic architecture underlying language performance is poorly understood. To identify novel genetic variants associated with language performance, we analyzed brain MRI and performed a genome-wide association study (GWAS) using a composite measure of language performance from the Alzheimer's Disease Neuroimaging Initiative (ADNI; n=1560). The language composite score was associated with brain atrophy on MRI in language and semantic areas. GWAS identified GLI3 (GLI family zinc finger 3) as significantly associated with language performance (p<5×10-8). Enrichment of GWAS association was identified in pathways related to nervous system development and glutamate receptor function and trafficking. Our results, which warrant further investigation in independent and larger cohorts, implicate GLI3, a developmental transcription factor involved in patterning brain structures, as a putative gene associated with language dysfunction in AD.