p120-catenin and β-catenin differentially regulate cadherin adhesive function.
Vascular endothelial (VE)-cadherin, the major adherens junction adhesion molecule in endothelial cells, interacts with p120-catenin and β-catenin through its cytoplasmic tail. However, the specific functional contributions of the catenins to the establishment of strong adhesion are not fully understood. Here we use bioengineering approaches to identify the roles of cadherin-catenin interactions in promoting strong cellular adhesion and the ability of the cells to spread on an adhesive surface. Our results demonstrate that the domain of VE-cadherin that binds to β-catenin is required for the establishment of strong steady-state adhesion strength. Surprisingly, p120 binding to the cadherin tail had no effect on the strength of adhesion when the available adhesive area was limited. Instead, the binding of VE-cadherin to p120 regulates adhesive contact area in a Rac1-dependent manner. These findings reveal that p120 and β-catenin have distinct but complementary roles in strengthening cadherin-mediated adhesion.


Vibrational spectrum, ab initio calculations, conformational stabilities and assignment of fundamentals of the Ci conformer of 1,4-dichlorobutane.
The infrared and Raman spectrum of 1,4-dichlorobutane is reported in solid, liquid and gas. Ab initio calculations for the nine stable or metastable conformers of 1,4-dichlorobutane are reported for Moller-Ploessett second order electron correlation and B3LYP density functionals with a variety of basis sets, using approximations as high as 6-311+g(2d, 2p). Normal coordinate calculations were conducted for the nine conformers and the results used to provide assignments for some of the observed infrared and Raman bands. An attempt to use the assignments together with the ab initio intensities or Raman activities to investigate the composition of the liquid at room temperature proved modestly successful, and suggested that the populations are altered from those expected in the gas phase by interactions of the permanent electric dipole moments with the dipolar plasma in which the conformers are immersed in the liquid. A substantial disagreement between the Moeller-Ploessett and density functional results is reported, and the calculation of intensities and activities is insufficiently accurate to allow detailed interpretation of the spectrum of the room temperature liquid. A complete assignment of fundamentals is given for the conformer of Ci symmetry, and one Raman and one infrared band is identified with the C2h conformer. All the other infrared and Raman bands in the liquid or the gas are composites of several contributors.


Prognostic value of serum MMP-8, -9 and TIMP-1 in patients with hepatocellular carcinoma.
Prediction of prognosis in hepatocellular carcinoma (HCC) is difficult. The aim of this study was to evaluate the prognostic value of serum MMP-8, -9, -13, and TIMP-1 in patients with HCC. Pre-treatment serum samples from 134 patients with HCC were retrospectively analyzed. The serum concentration of MMP-8 was analyzed with immunofluorometric assay (IFMA), and those of MMP-9, MMP-13, and TIMP-1 were determined by enzyme-linked immunosorbent assays (ELISA). Clinical data were retrieved from patient records and survival data obtained from Statistics Finland. The overall cumulative disease-specific survival was 69% at 1 year, 50% at 2 years, and 33% at 5 years. Kaplan-Meier overall survival analysis showed that patients with low concentrations of serum MMP-8 or TIMP-1 had a statistically significantly better overall survival than patients with high concentrations of serum MMP-8 or TIMP-1 (P=0.013 and P=0.003). Interestingly, the overall survival in patients with high MMP-9/TIMP-1 ratio was statistically significantly better than in those patients with low MMP-9/TIMP-1 ratio (P=0.004). Our results suggest that serum MMP-8, TIMP-1, and the ratio of MMP-9/TIMP-1 might be useful adjuncts as predictors of prognosis in patients with HCC.


Potential mechanisms of the acute coronary syndrome presentation in patients with the coronary slow flow phenomenon - insight from a plasma proteomic approach.
The coronary slow flow phenomenon [CSFP] is a coronary microvascular disorder, characterized by delayed distal vessel opacification despite the absence of obstructive coronary artery disease. Patients frequently present with an acute coronary syndrome [ACS] although the pathophysiological mechanisms responsible are unknown. The aim of this study was to identify potential mechanisms for the ACS presentation associated with the CSFP using a plasma proteomic profiling approach. Plasma samples from nine CSFP subjects [56 ± 11years] were assayed for high sensitivity C-reactive protein [hsCRP], troponin T [TnT], creatine kinase [CK], and proteomic analyses (n=6), during an ACS presentation and one month later [chronic phase]. Proteomic analysis involved chromatographic depletion of abundant plasma proteins followed by two-dimensional differential gel electrophoresis [2-D DIGE]. Protein spots demonstrating ±1.5-fold change relative to the control were identified by mass spectrometry and two differentially expressed proteins were selected for validation via Western blotting. During the ACS presentation, hsCRP was elevated [ACS=14.9 ± 3.9 mg/L vs chronic=4.23 ± 1.37 mg/L, p=0.05] but TnT and CK levels were unchanged. Proteomic analysis identified six proteins that were significantly different in abundance between the acute and chronic samples. During the ACS presentation there was a 1.6 ± 0.13 fold increase in the anti-oxidant enzyme paraoxonase-1 and an increase in inflammatory proteins alpha-1-antichymotrypsin [1.65 ± 0.13 fold] and alpha-1-antitrypsin [2.5 ± 0.34 fold]. The latter was confirmed by Western blotting [1.33 ± 0.17 OD acute/chronic ratio, p=0.05]. The findings from this novel detailed approach, implicate an inflammatory/oxidative stress process in the pathogenesis of the ACS presentation associated with the CSFP. Future studies should further elucidate these mechanisms.


Abdominal wall hernia repair with a composite ePTFE/polypropylene mesh: clinical outcome and quality of life in 152 patients.
No consensus has yet been reached regarding the optimal mesh for the repair of small ventral hernias. A composite polytetrafluoroethylene/polypropylene mesh (Ventralex(®)) is designed for this purpose, and this paper reports its use in a larger series of patients. Open repair for a small ventral hernia was undertaken in 152 patients between April 2006 and June 2008. Data from medical files were gathered, and follow-up questionnaires were retrieved. Patients were asked about pain, intake of analgesics and various physical capabilities. Patients with postoperative complaints were offered a follow-up visit. Ultrasonography was performed if recurrence was suspected. Median surgery time was 39 min (range 16-129 min). Junior surgeons performed 63% of the operations. Questionnaires were returned by 81.6% with a mean follow-up of 15.6 months. Eighteen patients (11.8%) had complications. Pain score was significantly lower and the physical capabilities of the patients were significantly enhanced after the operation. Recurrent hernia was reported in four patients (2.6%). Five patients (3.3%) had the mesh removed due to deep infection, chronic pain or early recurrence. The training level of the surgeon had no influence on the incidence of complications. A 93.8% majority of the patients would recommend this specific procedure to others. The intraperitoneal placement of this composite mesh is associated with a high level of patient satisfaction as well as low rates of both recurrence and infection.


Novel cytogenetic expression of gene amplification in actinomycin D-resistant somatic cell hybrids: transfer of resistance by centric chromatin bodies.
SEWATC13 mouse cells, resistant to 0.1 microgram/ml of actinomycin D (AMD), were fused to AMD-sensitive cells of the Chinese hamster ovary cell line (CHO). Twenty-two hybrid clones were isolated and put into serial culture in the selective medium. Unexpectedly, identifiable mouse chromosomes were found only in one of the hybrids. All the others had only hamster chromosomes and, in addition, numerous chromatin bodies (CBs), mostly small and irregularly shaped, but also larger, more chromosome-like ones. The CBs were distinctly C-band positive and a mouse satellite probe hybridized strongly to them. The AMD resistance of the murine parental cells had previously been attributed to gene amplification in two large homogeneously staining regions (HSR-AMD1 and 2). They were not observed in the hybrid cells but had supposedly reappeared in the guise of the CBs. It was established by Southern DNA blot analysis that amplified DNA sequences, localized to the HSR-AMD1 and 2 of the SEWA parent were present in multiple copies in the hybrids. It was also established by in situ hybridization that they were located in the CBs. Unlike double minutes (DMs) the CBs were all centric.


Actin-dependent activation of serum response factor in T cells by the viral oncoprotein tip.
Serum response factor (SRF) acts as a multifunctional transcription factor regulated by mutually exclusive interactions with ternary complex factors (TCFs) or myocardin-related transcription factors (MRTFs). Binding of Rho- and actin-regulated MRTF:SRF complexes to target gene promoters requires an SRF-binding site only, whereas MAPK-regulated TCF:SRF complexes in addition rely on flanking sequences present in the serum response element (SRE). Here, we report on the activation of an SRE luciferase reporter by Tip, the viral oncoprotein essentially contributing to human T-cell transformation by Herpesvirus saimiri. SRE activation in Tip-expressing Jurkat T cells could not be attributed to triggering of the MAPK pathway. Therefore, we further analyzed the contribution of MRTF complexes. Indeed, Tip also activated a reporter construct responsive to MRTF:SRF. Activation of this reporter was abrogated by overexpression of a dominant negative mutant of the MRTF-family member MAL. Moreover, enrichment of monomeric actin suppressed the Tip-induced reporter activity. Further upstream, the Rho-family GTPase Rac, was found to be required for MRTF:SRF reporter activation by Tip. Initiation of this pathway was strictly dependent on Tip's ability to interact with Lck and on the activity of this Src-family kinase. Independent of Tip, T-cell stimulation orchestrates Src-family kinase, MAPK and actin pathways to induce SRF. These findings establish actin-regulated transcription in human T cells and suggest its role in viral oncogenesis.


The effects of irradiation and microfiltration on the cells growing and total lipids production in the cultivation of Rhodotorula glutinis.
The results of this study indicate that the irradiation could enhance the cells growing of Rhodotorula glutinis to 54.2 ± 1.6g/L as compared to the control (without irradiation) of 38.3 ± 1.2g/L. However, different wavelength of LEDs' (red, green, blue and white) had no significant impacts on the growth and on the lipid content. The accumulation of potential inhibitive metabolic products probably impedes growth, which restricts more biomass accumulated in the fed-batch operation with irradiation. The combining of the fed-batch operation with irradiation and microfiltration can successfully improve the growth of R. glutinis to the maximum of 72.4 ± 0.6g/L and 51.2 ± 4.9% of lipid content obtained. Conclusively, the integration process of a fed-batch operation, irradiation and microfiltration can effectively enhance cell growth in R. glutinis, without any reimbursement of lipid contents. This finding might be useful when applied to the commercialized cultivation of R. glutinis for biodiesel production.


What is the probability of being too old for salvage transplantation after hepatocellular carcinoma resection?
The strategy of salvage transplantation for patients with hepatocellular carcinoma is based on the premise that tumour recurrence will be still transplantable at the time of recurrence. However, patients can not only present non-transplantable recurrence but can also be over the age limit accepted for transplantation. To measure the risk of being too old for salvage transplantation of patients resected for hepatocellular carcinoma within Milan criteria. A Markov simulation model was developed on the basis of published literature. The risk of being too old for salvage transplantation depends on the time-span between age at hepatic resection and age limit, and the expected median waiting-time. Patients resected at an age 2 or 3 years below the age limit carry a risk of being too old that overcomes the probability of receiving transplantation. Salvage strategy can cause harm that depends on the tumour characteristics and degree of portal hypertension, becoming maximal for patients with multiple tumours, clinical signs of portal hypertension and increased bilirubin levels. The best strategy to adopt should be balanced between the risk of being too old and the expected transplant benefit, but salvage strategy could be pursued if it did not turn into significant harm in comparison to primary transplantation.


Reassessing the species status of Pseudodiaptomus malayalus Wellershaus, 1969 and P. binghami Sewell, 1912 (Calanoida: Pseudodiaptomidae) from India based on morphology and mitochondrial cytochrome oxidase I gene sequences.
Pseudodiaptomus binghami Sewell, 1912 was first described from the Rangoon River (now Yangon River) estuary, Myanmar. Pseudodiaptomus malayalus Wellershaus, 1969 previously known as P. binghami malayalus, is a typical brackish-water calanoid copepod from Cochin Estuary, Kerala. Morphological examination of P. malayalus and P. binghami collected from Cochin Estuary and the Nambur canal in Andhra Pradesh revealed crucial differences between the two congeners. Female specimens of P. malayalus exhibited marked differences from those described by Wellershaus. They are (1) the number of terminal spines on P5, (2) ornamentation of GS, (3) ornamentation of Ur1-4, (4) length ratio of the Ur and CR segments and (5) length:width ratio of the CR setae. Furthermore, significant and discrete morphological differences were observed between the two Indian species in their P5 and urosome. But the male specimens of P. malayalus did not show any major differences from the original description. In addition, distance matrix data revealed 22% interspecific divergence values which in turn confirmed the status of P. malayalus and P. binghami as two distinct species.


Pacing for ventricular tachycardia.
Many problems remain to be solved before implanted pacers can assume a major role in the treatment of ventricular tachycardia. Case histories are cited to illustrate some of the difficulties to be overcome. Possible mechanisms for the success and failure of antitachycardia pacing are reviewed. Prospects for antitachycardia pacing will increase with the advent of cardioverter/defibrillator back-up.


Prolonged seizure suppression by a single implantable polymeric-TRH microdisk preparation.
Thyrotropin-releasing hormone (TRH; Protirelin) is an endogenous neuropeptide known to have anticonvulsant effects in several seizure models and in intractable epileptic patients. Like most neuropeptides, its duration of action may be limited by a lack of sustained site-specific bioavailability. To attempt to provide long-term delivery, we attached TRH to a biodegradable polyanhydride copolymer as a sustained-release carrier. Utilizing the rat kindling model of temporal lobe epilepsy, a single TRH microdisk implanted stereotaxically into the seizure focus (amygdala) significantly suppressed kindling expression when assessed by the number of stimulations required to reach each behavioral stage and to become fully kindled (8.63 +/- 0.92 vs. 16.17 +/- 1.37; Mean +/- S.E.M.). Two indices of seizure severity, afterdischarge duration (Mean +/- S.E.M., sec.) (stimulated amygdala [87.40 +/- 5.47 vs. 51.80 +/- 15.65] and unstimulated amygdala [89.60 +/- 5.55 vs. 48.67 +/- 15.8] and clonus duration (71.2 +/- 5.94 vs. 29.40 +/- 8.87; Mean +/- S.E.M., sec.), were also significantly reduced by a single polymeric-TRH implant. Fifty days after initiation of the study a significant reduction in clonus duration (53.90 +/- 3.27 vs. 40.09 +/- 4.14) still remained in the TRH-implanted groups. This report is the first to provide evidence in support of in situ microdisk pharmacotherapy for potential neuropeptide delivery in intractable epilepsy and possibly other neurological disorders.


Uncovering heterogeneity of anacardic acids from pistachio shells: A novel approach for structural characterization.
Anacardic acids (AnAs) are important secondary metabolites that occur primarily in plants of the Anacardiaceae family, such as pistachio (Pistacia vera L.). Some AnAs have been associated with health benefits, and the position of the CC double bonds is a crucial feature of these metabolites. Herein, we propose a new strategy based on RPLC separation and detection by ESI-MS/MS, preceded by an epoxidation reaction. The procedure was applied to the green extracts of lignified pistachio shells, and a mixture of AnAs bearing alkyl chains 13:0, 15:0, and 17:1 emerged as prevailing. As positional isomers of AnA 15:1 (Δ8 and Δ6) and AnAs 17:1 (Δ10 and Δ8) were identified for the first time, their discovery paves the way to the systematic study of their potential health-beneficial effects. The developed method was validated and applied to quantify AnAs in pistachio ethanolic extract, showing contents higher than 10 mg/ 100 g of biomass.


Effect of protein convertase subtilisin/kexin type 9 (PCSK9) 46L gene polymorphism on LDL cholesterol concentration in a Polish adult population.
The purpose was to study the effect of PCSK9 46L on cholesterol concentration and cardiovascular morbidity. By comparing 176 carriers with 6618 non-carriers identified through a cross-sectional population study (WOBASZ) we confirmed the LDL lowering effect of PCSK9 46L and demonstrated that it increases with the concentration of LDL. We noted that PCSK9 46L was associated with tendency for protection from myocardial infarction but not stroke suggesting a difference in the effect on susceptibility to these disorders.


Extended septoplasty for subaortic stenosis developed 19 years after double-outlet right ventricle repair.
A 27-year-old woman developed subaortic stenosis 19 years after double-outlet right ventricle repair. Subaortic stenosis was caused by a narrow ring of fibromuscular ridge associated with a bulge of the underlying septal muscle. The aortic valve was bicuspid and stenotic. We conducted extended septoplasty, replacing the aortic valve. Postoperative cineangiogram showed an adequate left ventricular outflow pathway. Double-outlet right ventricle repair may thus be followed by subaortic stenosis as long as 19 years after initial surgery. This lesion was assumed due to acquired disease secondary to flow disturbances in the left ventricular outflow, so reconstructing an adequate outflow pathway is effective and appears to help avoid recurring stenosis.


Targeted re-sequencing in pediatric and perinatal stroke.
Pediatric and perinatal stroke can present as an early symptom in undiagnosed syndromes characterized by simple Mendelian inheritance. In order to diagnose those patients affected with a monogenic disorder in which an arterial cerebrovascular event or arteriopathy may have preceded any other specific symptom, we aimed to establish and validate a targeted gene panel, and to determine its diagnostic yield and clinical utility. To this end, thirty-eight patients were selected with heterogeneous cryptogenic stroke phenotypes, mostly including multiple and recurrent ischemic or hemorrhagic arterial strokes and porencephalies, variably associated with calcifications, intracranial or systemic steno-occlusive arteriopathies, positive family history, and syndromic conditions. Clinical and neuroradiological data were collected for every patient enrolled in the study, and DNA samples were tested by means of a customized gene panel including 15 genes associated with known genetic diseases related to pediatric stroke. In four patients (10.5%) the analyses unraveled pathogenetic variants in ABCC6 and COL4A1 genes, leading to a definite genetic diagnosis with a great beneficial impact on patients management, while results were null in the remaining patients. These findings suggest a high complexity and variability of the included stroke phenotypes, that could not be fully accounted for by the genes tested in the present study. A wider gene panel or an unbiased genomic approach may be better suited and advisable to explain a greater proportion of pediatric and perinatal stroke events.


Anti-idiotypes and autoimmune disease.
Idiotypes represent structural sites located on the Fab fragment of antibodies. Even though these sites are combining sites for antigenic determinant, their uniqueness makes them antigenic, generating anti-Ids that have functionally performed as suppressive agents for the further production of the antibody. Jerne's network theory of idiotypes has provided a basis upon which investigators have searched for means to reduce, or explain the reduction of, Ab production. A principal area of this reduction of Ab expression concerns the autoAbs in autoimmune diseases. Experimental and clinical evidence suggests a role for anti-Ids in suppressing autoAb production and an inverse relationship between serum levels of anti-Ids and autoimmune disease activity.


Epidemiology and outcomes of acute respiratory failure in the United States, 2001 to 2009: a national survey.
The objective of this study was to evaluate trends in hospitalization, cost, and short-term outcomes in acute respiratory failure (ARF) between 2001 and 2009 in the United States. Using the Nationwide Inpatient Sample we identified cases of ARF based on International Classification for Diseases, Ninth Revision, Clinical Modification codes. We calculated weighted frequencies of ARF hospitalizations by year and estimated population-adjusted incidence and mortality rates. We used logistic regression to examine hospital mortality rates over time while adjusting for changes in demographic characteristics and comorbidities of patients. The number of hospitalizations with a diagnosis of ARF rose from 1,007,549 in 2001 to 1,917,910 in 2009, with an associated increase in total hospital costs from $30.1 billion to $54.3 billion. During the same period we observed a decrease in hospital mortality from 27.6% in 2001 to 20.6% in 2009, a slight decline in average length of stay from 7.8 days to 7.1 days, and no significant change in the mean cost per case ($15,900). Rates of mechanical ventilation (noninvasive [NIV] or invasive mechanical ventilation [IMV]) remained stable over the 9-year period, and the use of NIV increased from 4% in 2001 to 10% in 2009. Over the period of 2001 to 2009, there was a steady increase in the number of hospitalizations with a discharge diagnosis of ARF, with a decrease in inpatient mortality. There was a significant shift during this time toward the use of NIV, with a decrease in the rates of IMVuse.


Removal of middle molecules with medium cutoff dialyzer in patients on short frequent hemodialysis.
Medium cutoff (MCO) membranes for hemodialysis (HD) remove more effectively large middle molecules than high-flux (HF) membranes. In patients on in-center short frequent HD regimen (5 sessions per week, 2 hours and 30 minutes per session) the effect of MCO on middle weight uremic toxins has not been elucidated. This retrospective study included 15 patients previously performing short frequent HD with HF dialyzer (HF-HD), that were switched to short frequent HD with MCO dialyzer (MCO-HD) for 2 months, and returned to HF-HD. The primary endpoint was the predialysis concentration of α1-acid glycoprotein during the different study phases. Secondary endpoints were predialysis concentration of other middle molecules, albumin, and assessment of the quality of life using the 36-item short-form health survey (SF-36). During MCO-HD phase there was a reduction in mean ± SD α1-acid glycoprotein concentration (98.71 ± 25.2 vs. 88.6 ± 24.6 mg/dL, P = 0.107), followed by an increment 2 months after returning to HF-HD (89.18 ± 26.12 vs. 97.33 ± 31.29 mg/dL, P = 0.002); however, only the second variation was statistically significant. MCO-HD provided lower median predialysis concentration of prolactin (16 [10.2-25.6] vs. 14.1 [11.7-34.8] ng/mL, P = 0.036). Single-pool Kt/V, standard Kt/V, predialysis β2-microglobulin, myoglobin, and SF-36 questionnaire remained stable during the first two phases (pre-MCO and MCO). β2-Microglobulin increased in the post-MCO phase (20.02 ± 8.14 vs. 21.27 ± 7.64 μg/mL, P = 0.000). Mean predialysis concentration of albumin reduced significantly from pre-MCO vs. MCO phases (39.9 ± 3.7 vs. 38.3 ± 3.3 g/L, P = 0.020) and rebounded significantly from MCO vs. post-MCO phases (38.7 ± 3.1 vs. 41.3 ± 3.0 g/L, P = 0.007). In this retrospective analysis, short frequent MCO-HD promotes a reduction in prolactin, a middle weight uremic toxin, and trends toward a reduction in α1-acid glycoprotein. No patients developed hypoalbuminemia. These findings are encouraging and deserve investigation in prospective studies.


Identification of control elements 3' to the human keratin 1 gene that regulate cell type and differentiation-specific expression.
To define DNA regulatory elements that mediate the response of the keratin 1 (K1) gene to Ca(2+)-induced differentiation, regions spanning the 5'- and 3'-flanking sequences, coding regions, and introns from the human K1 gene were cloned into vectors containing the chloramphenicol acetyltransferase (CAT) reporter gene and transfected into cultured mouse keratinocytes. A 4.3-kilobase (kb) region located 3' to the K1 gene stimulated CAT activity in response to increasing Ca2+ concentrations from 0.05 mM (basal cells) to 1.2 mM (differentiated cells). The 4.3-kb fragment was also active in human epidermal cells but inactive in NIH 3T3 cells and primary mouse fibroblasts. Deletion analysis localized the activity to the terminal 1682 base pairs (bp) of the flanking sequence which retained Ca2+ sensitivity in epidermal cells but was not active in mesenchymal cells. Removal of a 207-base pair element created an enhancer which was active in both epidermal and mesenchymal cells but was still Ca(2+)-inducible. Further deletions identified two elements which functioned synergistically to give maximal Ca(2+)-sensitive activity. Stably transfected epidermal cell lines expressed CAT under the direction of these elements when grafted onto nude mice to reconstitute an intact epidermis. Previously reported keratin regulatory motifs were not contained in the 1682-bp fragment, but an AP-1 site was identified in one of the synergistic subunits.


Recent trends in the rural-urban suicide disparity among veterans using VA health care.
There is an elevated risk of suicide among people living in rural areas, and the rural-urban disparity in death by suicide is growing in the general United States population. The department of Veterans Affairs (VA) implemented programs targeting rural health in 2007 and suicide prevention in 2008. Rural-urban differences in suicide rates among VA users have not been examined since 2010. We sought to understand whether the rural-urban disparity in suicide risk among VA users decreased during a time of contemporaneous VA efforts to improve access to mental health care for rural Veterans and to improve the effectiveness of mental health services at preventing suicide. We performed a retrospective cohort study examining differences in the raw and adjusted annual suicide rate among rural and urban VA users between 2003 and 2017. All VHA users 2003-2017. Descriptive statistics are presented for all VHA users in 2017. This includes 6,120,355 unique VA users, 32.0% (n = 1,955,935) of whom lived at a rural address. Raw rates of death by suicide were higher in rural VA users than urban VA users overall (33.3 vs. 29.1 deaths per 100,000 population) and across years, but the age, sex, and race-adjusted rates converged in 2005. White VA users had over triple the rate of death by suicide as black VA users, and lived disproportionally in rural areas. The rural-urban suicide disparity among VA users persists. However, the disparity appears to be driven by differences in the racial composition of rural and urban patients, which were not accounted for in prior studies.


Purification and characterization of agarases from a marine bacterium Vibrio sp. F-6.
Marine bacterium Vibrio sp. F-6, utilizing agarose as a carbon source to produce agarases, was isolated from seawater samples taken from Qingdao, China. Two agarases (AG-a and AG-b) were purified to a homogeneity from the cultural supernatant of Vibrio sp. F-6 through ammonium sulfate precipitation, Q-Sepharose FF chromatography, and Sephacryl S-100 gel filtration. Molecular weights of agarases were estimated to be 54.0 kDa (AG-a) and 34.5 kDa (AG-b) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH values for AG-a and AG-b were about 7.0 and 9.0, respectively. AG-a was stable in the pH range of 4.0-9.0 and AG-b was stable in the pH range of 4.0-10.0. The optimum temperatures of AG-a and AG-b were 40 and 55 degrees C, respectively. AG-a was stable at temperature below 50 degrees C. AG-b was stable at temperature below 60 degrees C. Zn(2+), Mg(2+) or Ca(2+) increased AG-a activity, while Mn(2+), Cu(2+) or Ca(2+) increased AG-b activity. However, Ag(+), Hg(2+), Fe(3+), EDTA and SDS inhibited AG-a and AG-b activities. The main hydrolysates of agarose by AG-a were neoagarotetraose and neoagarohexaose. The main hydrolysates of agarose by AG-b were neoagarooctaose and neoagarohexaose. When the mixture of AG-a and AG-b were used, agarose was mainly degraded into neoagarobiose.


The influence of the definition of patient global assessment in assessment of disease activity according to the Disease Activity Score (DAS28) in rheumatoid arthritis.
Patient global assessment (PGA) is one of the 4 items included in the Disease Activity Score (DAS28) for evaluation of activity of rheumatoid arthritis (RA). We studied the influence of the use of 3 different techniques of PGA on the assessment of disease activity. We evaluated 3 different DAS28 according to the technique of PGA in 108 patients with active RA before and after 12 weeks of etanercept therapy. The reliability (intraclass coefficient of correlation) between screening and baseline was very high and similar for the 3 DAS28. The percentage of patients in the different states of disease (from remission to higher disease activity) and the sensitivity to change across the 3 DAS28 scales were very similar. The different techniques of collection of PGA to be included in the DAS calculation yield similar results. However, an accepted, unequivocal technique should be encouraged in order to reduce heterogeneity in scoring DAS among patients with RA.


Management of carotid artery stenosis. Update for family physicians.
To clarify the definition of carotid artery diseases, the appropriateness of screening for disease, investigation and management of patients presenting with transient ischemic attacks, and management of asymptomatic carotid bruits. SOURCES OF INFORMATION MEDLINE: was searched using the terms carotid endarterectomy, carotid disease, and carotid stenosis. Most studies offer level II or III evidence. Consensus statements and guidelines from various neurovascular societies were also consulted. Patients with symptoms of hemispheric transient ischemic attacks associated with >70% stenosis of the internal carotid artery are at highest risk of major stroke or death. Risk is greatest within 48 hours of symptom onset; patients should have urgent evaluation by a vascular surgeon for consideration of carotid endarterectomy (CEA). Patients with 50% to 69% stenosis might benefit from urgent surgical intervention depending on clinical features and associated comorbidity. Patients with <50% stenosis do not benefit from surgery. Asymptomatic patients with >60% stenosis should be considered for elective CEA. Symptomatic carotid artery syndromes need urgent carotid duplex evaluation to determine the need for urgent surgery. Those with the greatest degree of stenosis derive the greatest benefit from timely CEA.


Efficiency improvement of new Tetrathienoacene-based dyes by enhancing donor, acceptor and bridge units, a theoretical study.
A series of metal free Tetrathienoacene-based (TTA-based) organic dyes are designed and investigated as sensitizers for application in dye sensitized solar cells (DSSCs). Density function theory and time dependent density function theory calculations were performed on these dyes at vacuum and orthodichlorobenzene as the solvent. Effects of changing π-conjugation bridges and different functional groups in acceptor and donor units were investigated. UV-Vis absorption spectra were simulated to show the wavelength shifting and absorption properties. Inserting nitro and acyl chloride functional groups in acceptor and NH2 in donor units leads to the reduction of HOMO-LUMO gap by lowering the lowest unoccupied molecular orbital (LUMO) energy level and raising the highest occupied molecular orbital (HOMO) energy level and the increase in effective parameters in DSSC' efficiency. The results show that changing spacer units from thiophene to furan has a great effect on electronic structure and absorption spectra. Investigation of the electron distributions of frontier orbitals shows the HOMO and LUMO localization in donor and acceptor, respectively. Some key parameters that were studied here include light harvesting efficiency, free energy of electron injection and open circuit photo-voltage.


Early diagnosis of prostatic carcinoma: the role of transurethral resection.
The exclusion of prostatic carcinoma in TUR specimens represents a considerable problem for the histopathologist. A technique for the separation of tissue on an anatomical basis which has been carried out on 50 patients is presented. It permits more representative histological sampling of TUR specimens and more accurate characterisation of prostatic carcinoma, but further refinement is recommended.


Analysis of the upstream regulatory region of a chicken skeletal myosin heavy chain gene.
The organization of the cis-acting regulatory elements of a chick myosin heavy chain gene has been investigated. The data show that a gene which is transcribed in vivo in the fast white embryonic musculature is also the major transcript expressed during myotube differentiation of primary myoblasts derived from 12-day embryonic chick leg muscles. The upstream region of this gene consists of 7500 base pairs, and we have tested the ability of these sequences to drive expression of the chloramphenicol acetyltransferase gene in developing primary muscle cultures. Deletion analyses of the upstream region show that negative regulatory elements are present within 2000 base pairs of the basal promoter elements, the CCAAT and TAATA boxes. Removal of these elements reveals the presence of a strong positive element located near the start site of transcription. Sequence analysis showed that the region also contains a sequence characteristic of an enhancer found in the immunoglobulin heavy chain gene, ATGCAAAT, the "octa" element. Gel band-shift assays show that this octa sequence binds a transacting factor present in muscle nuclear extracts, although footprint analysis indicates a limited interaction. Transient assays carried out with a fragment in which the octa sequence has been mutated, with the subsequent abolition of protein binding, shows that the particular interaction probably plays a role in negatively modulating the action of the strong positive promoter element.


Surgical Treatment of Distal Biceps Ruptures.
Distal biceps ruptures occur from eccentric loading of a flexed elbow. Patients treated nonoperatively have substantial loss of strength in elbow flexion and forearm supination. Surgical approaches include 1-incision and 2-incision techniques. Advances in surgical technology have facilitated the popularity of single-incision techniques through a small anterior incision. Recently, there is increased focus on the detailed anatomy of the distal biceps insertion and the importance of anatomic repair in restoring forearm supination strength. Excellent outcomes are expected with early repair of the distal biceps, with restoration of strength and endurance to near-normal levels with minimal to no loss of motion.


A novel oligoribonuclease of Escherichia coli. I. Isolation and properties.
A new ribonuclease has been isolated from Escherichia coli. The enzyme is present in the 100,000 times g supernatant fraction and has been purified over 200-fold. Studies of the enzyme reveal that: 1. The enzyme shows a marked preference for oligoribonucleotides; indeed, the reaction rate is inversely proportional to the chain length of the substrate. The enzyme does not attack polynucleotides even at high concentrations of enzyme and has no detectable DNase activity. 2. The enzyme is stimulated strongly by Mn2+, less strongly by Mg2+, and not at all by Ca2+ and monovalent cations. 3. The enzyme is purified free of RNase I, RNase II, RNase III, polynucleotide phosphorylase, and other known ribonucleases of E. coli. The enzyme displays identical properties when isolated from mutants of E. coli that are deficient in the above ribonucleases. 4. The enzyme has a marked thermostability, a point of further distinction from RNase II.


Hapten-Ficoll conjugates induce T-cell-dependent IgM anti-hapten responses and T cells mediating hapten-specific delayed-type hypersensitivity.
It was generally believed until recently that the B-cell IgM response induced by type-II "thymus-independent" antigens, that are not mitogenic for B cells, was T cell independent. Neither has it been possible in the past to demonstrate that these antigens activate specific T cells. This has led to the general belief that type-II thymus-independent antigens have the anomalous property of not being able to activate such cells. Recent evidence has shown that the IgM response to hapten-Ficoll conjugates, the prototype of type-II thymus-independent antigens, is T cell dependent in at least some circumstances. Here evidence that these antigens activate T cells mediating hapten-specific delayed-type hypersensitivity (DTH) is presented. Furthermore, the conditions under which they do so are similar to those that allow thymus-dependent antigens to activate DTH-mediating T cells.


Potential of galvanotaxis to separation and cleaning of rumen ciliates.
The ability of rumen ciliate protozoa to move in a unidirectional electrical field from the anode to the cathode was tested in large-volume electromigration equipment; the aim was to concentrate the microorganisms and clean them of impurities. During galvanotaxis in the freshly harvested rumen fluid and at a voltage of 10 V (I=0.8 mA), cells of Isotricha (Isotricha prostoma, Isotricha intestinalis) were the first to swim towards the cathode; 1 min later, they were followed by Dasytricha ruminantium. Entodiniomorphous ciliates (small Entodiniae as well as large species) displayed minimum movement towards the cathode. The yield of electromigration of Entodinium caudatum from the in vitro culture ranged within 2-6% when using 75-100 V (10 mA) and 60 V (5 mA) voltage, respectively. At 10 V (0.8 mA), E. caudatum did not move towards the cathode. It is shown that the behavior of Trichostomatids (Dasytricha and Isotricha) facilitates separation by means of the tested large-volume equipment. Concentration and cleaning of Entodiniomorphous ciliates using galvanotaxis in the tested equipment proved to be ineffective.


A comparison of four different microleakage tests for assessment of leakage of root canal fillings.
The purpose of this study was to compare four different microleakage tests (dye leakage, electrochemical test, bacterial test and fluid filtration) for evaluation of the coronal seal of teeth obturated using Thermafil or lateral condensation techniques. One hundred forty-four extracted single-rooted teeth were selected and prepared. The specimens were divided into 8 experimental groups. In 4 groups the root canals were filled with Thermafil obturators and in the other 4 groups by the lateral condensation technique. Coronal microleakage was assessed after exposing one group of specimens (n = 15 teeth) from each of the obturation techniques to each of the four microleakage tests. In the fluid filtration test, lateral condensation showed statistically less leakage than the Thermafil technique (P < .05). Electrochemical and dye leakage test results showed no difference between the two obturation techniques (P < .05). However, in the bacterial leakage test, Thermafil showed less leakage than lateral condensation (P < .05). There is poor correlation between various methods to evaluate hydraulic leakage. The clinical significance of leakage tests in vitro is questionable.


A new procedure for coupling antibody to paper discs for radioimmunoassay: application to the determination of alpha-fetoprotein.
Horse anti-alpha-fetoprotein was coupled to CM-cellulose discs by a modified carbodiimide reaction. The resulting coupling CM-discs were used in solid-phase radioimmunoassay of human alpha-fetoprotein. The sensitivity of these discs and conventional BrCN activated filter paper discs coupled anti-alpha-fetoprotein was approximately the same. A fair correlation between the alpha-fetoprotein levels determined by both methods was observed. The coupling procedure with carbodiimide is simple and the use of hazardous BrCN is eliminated.


Meta-analysis of genome-wide association scans for genetic susceptibility to endometriosis in Japanese population.
To identify susceptibility genes for endometriosis in Japanese women, genome-wide association (GWA) analysis was performed using two case-control cohorts genotyped with the Affymetrix Mapping 500K Array or Genome-Wide Human SNP Array 6.0. In each of the two array cohorts, stringent quality control (QC) filters were applied to newly obtained genotype data, together with previously analyzed data from the Japanese Integrated Database Project. After QC-based filtering of samples and single nucleotide polymorphisms (SNPs) in each cohort, 282 838 SNPs in both genotyping platforms were tested for association with endometriosis using a meta-analysis of the two GWA studies with 696 patients with endometriosis and 825 controls. The meta-analysis revealed that a common susceptibility locus conferring a large effect on the disease risk was unlikely. On the other hand, an excess of SNPs with P-values <10(-4) (36 vs 28 SNPs expected by chance) was observed in the meta-analysis. Of note, four of the top five SNPs with P-values <10(-5) were located in and around IL1A (interleukin 1α), which might be a functional candidate gene for endometriosis. Further studies with larger case-control cohorts will be necessary to elucidate the genetic risk factors.


Alterations of benzodiazepine receptor binding in tremor rats with absence-like seizures.
Tremor rats begin to exhibit clinical or electrical absence-like seizures after 6 weeks of age, and by 14 weeks of age, all have seizures. Central-type benzodiazepine receptor binding was investigated in tremor rats and control rats, aged 4 weeks and 16 weeks. Significantly lower benzodiazepine receptor density and no differences in affinity were found in the hippocampus of the tremor rats in comparison with that of control rats at both ages. This abnormality is considered to be due to a tremor gene and may be the cause of absence-like seizures in tremor rats. A significantly lower receptor density was found in the cerebellum at 4 weeks of age in the tremor rats than in the control rats. These changes may be related to tremorous movements in the tremor rats. Receptor density was significantly lower in the brainstems of tremor rats and control rats at 16 weeks of age than at 4 weeks of age, and the decrease was more marked in control rats. These facts may reflect a reduced decrease in the response to the dysfunction of gamma-aminobutyric acidergic neurons, or the function of the gamma-aminobutyric acid/benzodiazepine receptor system may be secondarily increased to suppress seizures in 16-week-old tremor rats.


Limited agreement exists between rationale and practice in athletes' supplement use for maintenance of health: a retrospective study.
The widespread use of nutritional supplements among athletes is poorly understood. The prevalence of supplement intake and users' knowledge have been researched independently leading to useful, but disconnected, information on supplement use. The 'UK Sport 2005 Drug Free Survey' data (n = 874) were re-analysed using association [chi2] and 'strength of association' tests [phi], to discover observed incongruencies between self-reported supplement use and the underlying motives. Results are given for test pairs between 'motive for use' [doctor's advice, avoiding sickness, overcoming injuries and enhancement of diet] and each supplement used and these were categorized as strong (phi > .7), intermediate (7 < phi > .3) and weak (phi < .3). The use of selected supplements varied widely as follows: multivitamin (72.7%), vitamin C (70.4%), echinacea (30.8%), iron (29.8%), magnesium (11.0%) and ginseng (8.3%). Associations with motive were found in 8 of the 10 test pairs which were expected from literature precedents, however only weak associations exist. Of these, four were associated with avoidance of sickness [iron (chi2 = 11.94, p < .001; phi = .15, p = .001), multivitamin (chi2 = 6.43, p < .001; phi = .11, p = .011), vitamin C (chi2 = 54.67, p < .001; phi = .32, p < .001) and echinacea (chi2 = 40.34, p < .001; phi = .28, p < .001)]. The remaining 4 associations were: no time to prepare meals with ginseng (chi2 = 7.64, p = .006; phi = .12, p = .006) and multivitamin (chi2 = 9.103, p = .003; phi = .13, p = .003); overcoming injuries with magnesium (chi2 = 6.99, p = .008; phi = .11, p = .008); doctors' advice and iron (chi2 = 35.00, p < .001; phi = .25, p = .001). These results suggest a lack of understanding regarding supplements and health maintenance, except for vitamin C and echinacea. Furthermore, supplement use is apparently independent of physicians/dieticians' advice, except for iron. This may suggest a widespread circumvention of expert advice in the growing area of supplement use and therefore should be addressed to underscore potential health risks.


PCR for varicella zoster virus genome negative in corneal epithelial cells of patients with Thygeson's superficial punctate keratitis.
To find out if varicella zoster virus is the causative agent of Thygeson's superficial punctuate keratitis. Epithelial cells were harvested from the punctuate epithelial lesions 9 patients with Thygeson's superficial punctate keratitis. After DNA extraction polymerase chain reaction was carried out with varicella zoster virus primers. All samples were negative with regard to varicella zoster virus genome. This result suggests that varicella zoster virus is most probably not the causative agent of Thygeson's superficial punctate keratitis.


Ion movement through gramicidin A channels. Single-channel measurements at very high potentials.
The patch-clamp technique of Mueller (1975, Ann. N.Y. Acad. Sci., 274:247-264) and Neher and Sakmann (1976, Nature (Lond.), 260:799-802) was modified to be suitable for single-channel measurements in lipid bilayers at potentials up to 500 mV. This method was used to study gramicidin A single-channel current-voltage characteristics. It was found that the sublinear current-voltage behavior normally observed at low permeant ion concentrations and rather low potentials (V less than or equal to 200 mV) continues to be seen all the way up to 500 mV. This phenomenon is characteristic of the low permeant ion situation in which the channel is far from saturation, and implies that the overall rate constant for association between ion and channel is very weakly, if at all, voltage dependent. The magnitude of the single channel currents at 500 mV is consistent with the notion that the aqueous convergence conductance is a significant factor in determining the permeability characteristics of the gramicidin A channel.


Resolving protein-mineral interfacial interactions during in vitro mineralization by atom probe tomography.
Organic macromolecules exert remarkable control over the nucleation and growth of inorganic crystallites during (bio)mineralization, as exemplified during enamel formation where the protein amelogenin regulates the formation of hydroxyapatite (HAP). However, it is poorly understood how fundamental processes at the organic-inorganic interface, such as protein adsorption and/or incorporation into minerals, regulates nucleation and crystal growth due to technical challenges in observing and characterizing mineral-bound organics at high-resolution. Here, atom probe tomography techniques were developed and applied to characterize amelogenin-mineralized HAP particles in vitro, revealing distinct organic-inorganic interfacial structures and processes at the nanoscale. Specifically, visualization of amelogenin across the mineralized particulate demonstrates protein can become entrapped during HAP crystal aggregation and fusion. Identification of protein signatures and structural interpretations were further supported by standards analyses, i.e., defined HAP surfaces with and without amelogenin adsorbed. These findings represent a significant advance in the characterization of interfacial structures and, more so, interpretation of fundamental organic-inorganic processes and mechanisms influencing crystal growth. Ultimately, this approach can be broadly applied to inform how potentially unique and diverse organic-inorganic interactions at different stages regulates the growth and evolution of various biominerals.


Identification of glycopeptide-resistant enterococci by VITEK 2 system and conventional and real-time polymerase chain reaction.
Glycopeptide-resistant enterococci (GRE) are important causes of nosocomial infections in the United States and in Europe. Rapid detection of GRE is essential for the implementation of appropriate control measures to prevent the spread of GRE. In this study, we compared the reliability of 3 different methods, VITEK 2 automated system (bioMérieux), a conventional multiplex polymerase chain reaction (m-PCR) protocol, and a real-time PCR protocol performed on the LightCycler system (Roche) for identification of GRE in the routine microbiology laboratory. Species identification and glycopeptide resistance determination was tested with 80 enterococcal isolates with different glycopeptide resistance phenotypes. With the VITEK 2 system, 39% of the strains were correctly identified to species level. Resistance to vancomycin was detected in all isolates; however, discrepancies occurred in the correct detection of teicoplanin resistance. The PCR protocols proved to be suitable for detecting clinically relevant GRE; 90% of the isolates studied were correctly identified with the conventional m-PCR and 100% of vanA and vanB isolates with the real-time PCR protocol, respectively. High specificity and rapidity make the real-time PCR assays superior tools for identification of GRE in clinical samples; however, they do not have the ability to detect vanC.


A new technique for internal fixation of femoral fractures in mice: impact of stability on fracture healing.
Mouse models are of increasing interest to study the molecular aspects of fracture healing. Because biomechanical factors greatly influence the healing process, stable fixation of the fracture is of interest also in mouse models. Unlike in large animals, however, there is a lack of mouse models which provide stable osteosynthesis. The purpose of this study was therefore to develop a technique for a more stable fixation of femoral fractures in mice and to analyze the impact of stability on the process of fracture healing. The new technique introduced herein includes an intramedullary pin and an extramedullary metallic clip. Ex vivo biomechanical analysis revealed a significantly higher implant stiffness of our pin-clip technique when compared with previously described intramedullary fixation techniques. In vivo, we studied the course of healing after the more stable fixation with our pin-clip technique and compared the results with that observed after unstable fixation with the pin-clip technique after cutting the clip. After 2 and 5 weeks of fracture healing radiological analysis demonstrated that the more stable fixation with the pin-clip technique results in a significantly higher union rate compared to the unstable fixation. Torsional stiffness at 5 weeks was almost 3-fold of that measured after unstable fixation. Histomorphological analysis further showed that fractures stabilized with the pin-clip technique healed with a smaller periosteal callus area, an increased fraction of bone and a reduced amount of fibrous tissue. Of interest, the pin-clip fixation showed reliable union after 5 weeks, whereas the unstable pin fixation did not regularly achieve adequate fracture healing. In conclusion, we introduce a novel, easily applicable internal osteosynthesis technique in mice, which provides rotational stability after femoral fracture fixation. We further show that a more stable osteosynthesis significantly improves the process of fracture healing also in mice.


Efficacy and safety of vixotrigine in idiopathic or diabetes-associated painful small fibre neuropathy (CONVEY): a phase 2 placebo-controlled enriched-enrolment randomised withdrawal study.
No pharmacological treatments are specifically indicated for painful small fibre neuropathy (SFN). CONVEY, a phase 2 enriched-enrolment study, evaluated the efficacy and safety of vixotrigine, a voltage- and use-dependent sodium channel blocker, in participants with idiopathic or diabetes-associated painful SFN. CONVEY was a phase 2, multicentre, placebo-controlled, double-blind (DB), enriched-enrolment, randomised withdrawal study. The study was conducted at 68 sites in 13 countries (Europe and Canada) between May 17, 2018, and April 12, 2021. Following a 4-week open-label period in which 265 adults with painful SFN (a mixture of large and small fibre neuropathy was not exclusionary) received oral vixotrigine 350 mg twice daily (BID), 123 participants (with a ≥30% reduction from baseline in average daily pain [ADP] score during the open-label period) were randomised 1:1:1 to receive 200 mg BID, 350 mg BID or placebo for a 12-week double-blind (DB) period. Primary endpoint was change from baseline in ADP at DB Week 12. Secondary endpoints included the proportion of participants with a ≥30% reduction from baseline in ADP and the proportion of Patient Global Impression of Pain (PGIC) responders at DB Week 12. Treatment-emergent adverse events (AEs) were monitored. Statistical significance was set at 0.10 (2-sided). The trial was registered on ClinicalTrials.gov (NCT03339336) and on ClinicalTrialsregister.eu (2017-000991-27). A statistically significant difference from placebo in least squares mean reduction in ADP score from baseline to DB Week 12 was observed with vixotrigine 200 mg BID (-0.85; SE, 0.43; 95% CI, -1.71 to 0.00; p = 0.050) but not 350 mg BID (-0.17; SE, 0.43; 95% CI, -1.01 to 0.68; p = 0.70). Numerically, but not statistically significantly, more participants who received vixotrigine vs placebo experienced a ≥30% ADP reduction from baseline (68.3-72.5% vs 52.5%), and only the 350 mg BID group had significantly more PGIC responders vs placebo (48.8% vs 30.0%; odds ratio = 2.60; 95% CI, 0.97-6.99; p = 0.058) at DB Week 12. AEs were mostly mild to moderate in the vixotrigine groups. The most common AEs (≥5% of vixotrigine-treated participants) in the DB 200 mg BID and 350 mg BID vixotrigine groups were falls, nasopharyngitis, muscle spasm, and urinary tract infection. In our study, vixotrigine 200 mg BID, but not 350 mg BID, met the primary endpoint; more vixotrigine-treated participants experienced a ≥30% reduction from baseline in ADP at DB Week 12. Vixotrigine (at both dosages) was well tolerated in participants with SFN. Biogen, Inc.


Severe hypotension from epidural meperidine in a high-risk patient after thoracotomy.
A sixty-eight-year-old female developed severe hypotension immediately after the administration of epidural meperidine for post-thoracotomy pain. Two preceding injections of epidural opiates had been uneventful. The cardiovascular collapse was difficult to reverse and may have contributed to the patient's subsequent death. Cardiovascular complications have not been reported frequently as a possible side effect of epidural opiate analgesia.


Elevated serum α-fetoprotein levels in patients with chronic hepatitis C virus genotype 4: not the end of the story.
Elevated serum α-fetoprotein (AFP) is not uncommonly seen among patients with chronic hepatitis C. This study aimed to identify clinical characteristics, histological characteristics, and biochemical markers associated with increased serum AFP levels in hepatitis C virus genotype 4-infected patients with no evidence of hepatocellular carcinoma and to determine the effect of lifestyle modification on these parameters. The study included 447 chronic hepatitis C patients with no evidence of hepatocellular carcinoma and 100 healthy controls. They underwent liver biopsies, homeostasis model assessment-insulin resistance (HOMA-IR), measurement of serum insulin, leptin, adiponectin, tumor necrosis factor-α, and interleukin-6 levels by an enzyme-linked immunosorbent assay, and assessment of AFP levels. Eighty patients with HOMA-IR greater than 3 received prospective longitudinal lifestyle intervention. In a multivariate analysis, platelet count less than 140×10/cm, a mean platelet volume of at least 9.5 fl, a neutrophil-lymphocyte ratio (NLR) of at least 2, an aspartate transaminase level of at least 55 IU/l, a γ-glutamyl transpeptidase level of at least 40 IU/l, an albumin level of up to 3.8 g/dl, HOMA-IR greater than 3, a leptin level of at least 10 pg/ml, an iron level of at least 165 μg/dl, a ferritin level of at level 175 ng/ml, and hepatic fibrosis F3-F4 were found to be independently associated with elevated AFP levels. The lifestyle intervention significantly improved BMI, platelet indices, NLR, γ-glutamyl transpeptidase, leptin, leptin/adiponectin ratio, tumor necrosis factor-α, interleukin-6, HOMA-IR, and AFP levels. Elevated insulin resistance, leptin, serum iron, ferritin, mean platelet volume, NLR, and advanced fibrosis, as well as decreased platelet count and serum albumin, are independently associated with an elevated AFP level. Lifestyle modification can improve (reduce) insulin resistance, leptin, leptin/adiponectin ratio, platelet count and their indices, NLR, and AFP level.


Molecular basis of weak D in Taiwanese.
Two genes, RHD and RHCE, encode the antigens of the RH blood group system. The weak D phenotype is caused by many different RHD alleles encoding aberrant RhD proteins, resulting in distinct serologic phenotypes and anti-D immunization. We analyzed seven weak D phenotypes excluding D(el), using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and direct sequencing methods to detect the changes of all ten RHD exons. The results show that there are four types of weak D in Taiwanese: one case each for CGG to CAG mutation at codon 10, GTG to ATG mutation at codon 174, and GTG to GAG mutation at codon 270, and four cases for GGT to GAT mutation at codon 282. In conclusion, we present the first data of a molecular basis of weak D in Taiwanese, which suggest a clinically relevant potential for anti-D immunization and may improve transfusion strategy in weak D Taiwanese patients.


[Endo-microscopic characteristics of the superficial epithelium in patients with cancer of the large intestine].
In vivo studies of the large bowel mucosa in 127 patients, using colon microscopy, showed the superficial epithelium to have a heterogeneous structure comprising three major types of cellular patterns. There is a relationship between said patterns and morphological structure in the most frequent neoplasms of the large bowel.


Disagreement in normative IGF-I levels may lead to different clinical interpretations and GH dose adjustments in GH deficiency.
Normative data for the iSYS IGF-I assay have been published both in the VARIETE cohort and by Bidlingmaier et al. To investigate whether normative data of the VARIETE cohort lead to differences in Z-scores for total IGF-I and clinical interpretation compared to normative data of Bidlingmaier et al. We used total IGF-I values previously measured by the IDS-iSYS assay in 102 GH-deficient subjects before starting GH treatment and after 12 months of GH treatment. Z-scores were calculated for all samples by using the normative data of the VARIETE cohort and by the normative data reported by Bidlingmaier et al. Before GH treatment, Z-scores calculated by using the normative data of the VARIETE cohort were significantly lower than those calculated by the normative data of Bidlingmaier et al: -2.40 (-4.52 to +1.31) (mean [range]) vs. -1.41 (-3.14 to +1.76); P < .001). After 12 months of GH treatment, again the Z-scores based on the normative data of the VARIETE cohort were significantly lower than those based on the normative data of Bidlingmaier et al: -0.65 (-4.32 to +2.79) vs 0.21 (-3.00 to +3.28); P < .001). IGF-I Z-scores in 102 GH-deficient subjects differed significantly when normative data from two different sources were used. In daily clinical practice, this would most likely have led to different clinical interpretations and GH dose adjustments.


Expression of nonconscious knowledge via ideomotor actions.
Ideomotor actions are behaviours that are unconsciously initiated and express a thought rather than a response to a sensory stimulus. The question examined here is whether ideomotor actions can also express nonconscious knowledge. We investigated this via the use of implicit long-term semantic memory, which is not available to conscious recall. We compared accuracy of answers to yes/no questions using both volitional report and ideomotor response (Ouija board response). Results show that when participants believed they knew the answer, responses in the two modalities were similar. But when they believed they were guessing, accuracy was at chance for volitional report (50%), but significantly higher for Ouija response (65%). These results indicate that implicit semantic memory can be expressed through ideomotor actions. They also suggest that this approach can provide an interesting new methodology for studying implicit processes in cognition.


The distribution of polychaetes on submerged Cystoseira has been studied, mainly in the sublitoral from 0 to 2.5 meters, and at localities of different exposure. 1. 93 species belonging to 18 different families were found. Algae with no epiphytes are colonized by only a few species of polychaetes most of which can also be found in other habitats. 2. In order to develop most polychaetes require a secondary structure such as is provided by the epiphytes growing on the cystoseiras and by epizoic animals. One fifth of the species of polychaetes known from the Mediterranean occurs in the epigrowth of submerged Cystoseira. 3. The structure of Cystoseira specimen is conditioned by hydrodynamic factors of the medium. There is a denser population of polychaetes in exposed areas. The algae growing on cystoseiras can be divided into ekto-, meso- and basosessile species, and according to their degree of dependence on algal structure the polychaetes may be epi-, meso- or endobionts. 4. The conclusion to be drawn from this investigation is that most polychaetes occurring on cystoseiras need only small interstitial spaces to develop and have no specific food requirements.


Dephosphorylation of the focal adhesion protein VASP in vitro and in intact human platelets.
The focal adhesion protein VASP, a possible link between signal transduction pathways and the microfilament system, is phosphorylated by both cAMP- and cGMP-dependent protein kinases in vitro and in intact cells. Here, the analysis of VASP dephosphorylation by the serine/threonine protein phosphatases (PP) PP1, PP2A, PP2B and PP2C in vitro is reported. The phosphatases differed in their selectivity with respect to the dephosphorylation of individual VASP phosphorylation sites. Incubation of human platelets with okadaic acid, a potent inhibitor of PP1 and PP2A, caused the accumulation of phosphorylated VASP indicating that the phosphorylation status of VASP in intact cells is regulated to a major extent by serine/threonine protein phosphatases. Furthermore, the accumulation of phosphorylated cAMP-dependent protein kinase substrate(s) appears to account for inhibitory effects of okadaic acid on platelet function.


Next generation genome sequencing reveals phylogenetic clades with different level of virulence among Salmonella Typhimurium clinical human isolates in Hong Kong.
Salmonella Typhimurium is frequently isolated from foodborne infection cases in Hong Kong, but the lack of genome sequences has hindered in-depth epidemiological and phylogenetic studies. In this study, we sought to reconstruct the phylogenetic relationship and investigate the distribution and mutation patterns of virulence determinants among local S. Typhimurium clinical isolates using their genome sequences. We obtained genome sequences of 20 S. Typhimurium clinical isolates from a local hospital cluster using a 454 GS FLX Titanium sequencing platform. Phylogenetic analysis was performed based on single nucleotide polymorphism positions of the core genome against the reference strain LT2. Antimicrobial susceptibility was determined using minimal inhibitory concentration for five antimicrobial agents and analyses of virulence determinants were performed through referencing to various databases. Through phylogenetic analysis, we revealed two distinct clades of S. Typhimurium isolates and three outliers in Hong Kong, which differ remarkably in antimicrobial susceptibility and presentation and mutations of virulence determinants. The local isolates were not closely related to many of the previously sequenced S. Typhimurium isolates, except LT2. As the isolates in the two clades spanned over 10 years of isolation, they probably represent endemic strains. The outliers are possibly introduced from outside of Hong Kong. The close relatedness of members in one of the clades to LT2 and the Japanese stool isolate T000240 suggests the potential reemergence of LT2 progeny in regions nearby. Our study demonstrated the utility of next-generation sequencing coupled to traditional microbiological testing method in a retrospective epidemiological study involving multiple clinical isolates. The evolution of multidrug- and ciprofloxacin-resistant strains among the more virulent clade is also an increasing concern.


Osteomalacia Prevalence, Biochemical Profile, and Histology in Patients with Low-Energy Hip Fractures Over the Age of 45.
Our objective was to determine the prevalence of osteomalacia in low-energy hip fracture patients over the age of 45, based on biochemical and histological measures. This cross-sectional study included 72 patients over 45 with low-energy mechanism hip fractures. Samples of fasting venous blood were taken for hemograms and serum biochemistry analyses. Bicortical biopsies of the iliac crest were obtained, processed, and evaluated by an expert pathologist for osteomalacia. Biochemical osteomalacia (b-OM) is defined according to a distinct criterion. A low level of serum calcium, phosphorus, albumin, and 25OHD was found in 43.1, 16.7, 73.6, and 59.7% of patients, respectively. 50.0% of patients had high serum alkaline phosphatase (ALP) levels. b-OM was found in 30 (41.7%), and no significant association was found with PTH, Cr, Alb, age, sex, fracture type, side of the trauma, and season were not associated with osteomalacia. Osteomalacia was diagnosed on histopathological analysis in 19/72 (26.7%), and 54/72 (75.0%) of all cases fulfilled b-OM criteria. In the histologic evaluation, osteoid seam width, osteoid surface, and osteoid volume were 28.5 µm, 25.6, and 12.1%, respectively. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the biochemical test for detecting osteomalacia were 73.6, 64.2, 42.4, 87.2, and 66.7%, respectively. Up to 30% of elderly patients with low-energy hip fractures are affected by osteomalacia. A biochemical screening along with a bone biopsy and histopathologic evaluation may be logical in a high-risk population for osteomalacia diagnosis.


Metamemory in patients with schizophrenia measured by the feeling of knowing.
Cognitive awareness in patients with schizophrenia is crucial for clinical management of cognitive deficits. Traditional approaches using self-report inventories have questionable validity and reliability. Using the Feeling-of-Knowing (FOK) procedure to measure metamemory might overcome such weakness. A cross-sectional study comparing 40 patients and 40 demographically matched normal controls, using a recall-judgment-recognition (RJR) procedure and the Hamann coefficient was conducted to examine whether patients with schizophrenia have FOK deficits and what neurocognitive mechanism might account for these deficits. General IQ, executive function, and memory tests were also assessed. The results show that as a group, patients with schizophrenia had impaired FOK ability and evidenced a disposition to underestimate their memory performance. However, patient's FOK ability was variable, with 42.5% of patients exhibited a below chance level performance. There were marked relationships between FOK and set formation and visual recognition abilities in healthy controls, while such feature was not evident in patient group. These findings are in line with the prefrontal cortex dysfunction, reduced intrapsychic monitoring ability, and impaired utilization of mental resources noticed in patients with schizophrenia. A routine evaluation of metamemory function by FOK might be helpful for designing customized cognitive rehabilitation programs considering their missed estimation of memory capacity.


Comparison of two ruminoscopy techniques in calves.
Two different techniques were tested in nine calves to describe endoscopically the visible structures of the rumen and the reticulum. Ruminoscopy using an orally introduced flexible endoscope proved to be unsatisfactory, but when the endoscope was introduced through a ruminal fistula in the left flank into the dorsal and ventral ruminal sacs, the blind sacs, ruminal pillars, the atrium of the rumen, the reticulum and the reticular groove could all be displayed effectively. Indentations of the ruminal wall caused by the left kidney, the spleen and the abomasum could be seen with the endoscope as could reticulo-ruminal contractions. The technique allowed visual evaluation of the mucosal surfaces and movement of the rumen, reticulum and reticular groove. The advantages and disadvantages of ruminoscopy in bovine medicine are discussed.


Validation of subjective rating scales for assessment of surgical workspace during laparoscopy.
Recently, studies have focused on how to optimize laparoscopic surgical workspace by changes in intra-abdominal pressure, level of muscle relaxation or body position, typically evaluated by surgeons using subjective rating scales. We aimed to validate two rating scales by having surgeons assess surgical workspace in video sequences recorded during laparoscopic surgery. Video sequences were obtained from laparoscopic procedures. Eight experienced surgeons assessed the video sequences on a categorical 5-point scale and a numerical 10-point rating scale. Intraclass correlations coefficients (ICC) and 95% confidence intervals (CI) were calculated for intra- and inter-rater reliability. The 5-point rating scale had an intra-rater ICC of 0.76 (0.69; 0.83) and an inter-rater ICC of 0.57 (0.45; 0.68), corresponding to excellent and fair reliability, respectively. The 10-point scale had an intra-rater ICC of 0.86 (0.82; 0.89) and an inter-rater ICC of 0.54 (0.39; 0.68), corresponding to excellent and fair as well. All surgeons used the full range of the 5-point scale, but only one surgeon used the full range of the 10-point scale. In conclusion, both scales showed excellent intra-rater and fair inter-rater reliability for assessing surgical workspace in laparoscopy. The 5-point surgical rating scale had all categories employed by all surgeons.


[Clinical value of the colored contrast sensitivity test. Preliminary note].
Evaluation of color contrast sensitivity is a new test for the assessment of visual capacity and especially of light stimulation transfer along the optic pathway. The color technique yields more accurate results. It is combined with previously existing functional examinations and allows very early detection of subclinical signs of starting pathological processes.


Immunohistochemical identification of alpha-1-antitrypsin, alpha-1-antichymotrypsin, and lysozyme in focal hyperplastic gingivitis.
Tissues from twenty-four patients with focal hyperplastic gingival lesions containing calcification were stained for lysozyme (muramidase), alpha-1-antitrypsin, and alpha-1-antichymotrypsin. In eighteen of the twenty-four cases the tissues stained positively for lysozyme, and in all instances the tissues stained positively for alpha-1-antitrypsin and alpha-1-antichymotrypsin. These data suggest that the fibrous components of these lesions are derived from tissue histiocytes.


Bradyarrhythmias in patients with COVID-19: A case series.
Novel coronavirus disease (COVID-19) can have variety of cardiac manifestations; however, less is known about the prevalence, clinical characteristics and outcomes of bradyarrhythmias in patients with COVID-19. In the present case series of bradyarrhythmia in patients with COVID-19, we report complete heart block requiring intervention in 5 patients and sinus node dysfunction in 2 patients.


Photocatalytic degradation of metronidazole and oxytetracycline by novel l-Arginine (C, N codoped)-TiO2/g-C3N4: RSM optimization, photodegradation mechanism, biodegradability evaluation.
Removal of Metronidazole (MNZ) and Oxytetracycline (OTC) from wastewater by the prepared (C, N codoped)-TiO2/g-C3N4 (Graphitic carbon nitride) was examined. l-Arginine (C, N codoped)-TiO2 and l-Arginine (C, N codoped)-TiO2/g-C3N4 photocatalysts were successfully synthesized through the sol-gel method, and optimal ratio of l-arginine:TiO2, as well as l-arginine/TiO2:g-C3N4, was determined by a kinetic study of photodegradation process. The maximum photocatalytic removal rate (0.062 min-1 for MNZ removal) was observed using 1% l-Arginine-TiO2/g-C3N4 (1:1) under visible light illumination, 2.2 and 12.4 times greater than those of 1% l-Arginine-TiO2 and pure TiO2, respectively. l-Arginine (1%)-TiO2/g-C3N4 (1:1) (co-doped-TCN) was investigated using X-ray diffraction analysis (XRD), Field Emission Scanning Electron Microscopy (FESEM), Energy Dispersive X-ray (EDX), Photo-luminescence (PL), and Differential Reflectance Spectroscopy (DRS) as the best-performing photocatalyst. Response surface methodology (RSM) was used to study the effect of co-doped-TCN dosage (0.5-1.0 g/L), pH of simulated wastewater (4-10), initial concentration of MNZ and OTC (50-100 mg/L), and irradiation time (30-90 min for MNZ and 20-40 min for OTC) on removal efficiency of the antibiotics. Also, their optimum values were determined by RSM. The treated pharmaceutical wastewater showed high biodegradability features with 5-day biological oxygen demand/chemical oxygen demand (BOD5/COD) of 0.51 and 0.46 after 40 and 100 min reaction for OTC and MNZ, respectively. The order of reactive species responsible for the photodegradation of pollutants was •O2─> •OH > h+>1O2. The effect of inorganic anions showed that all anions decreased the removal efficiency of both antibiotics in order of NO3─> Cl─ >SO42─>HPO42─ >HCO3─ for MNZ and NO3─> SO42─ > Cl─ >HPO42─ >HCO3─ for OTC. Also, introducing different oxidants improved the photocatalytic removal efficiency with the order of H2O2>K2S2O8> KBrO3.


Three-dimensional atomic-scale observation of structural evolution of cathode material in a working all-solid-state battery.
Most technologically important electrode materials for lithium-ion batteries are essentially lithium ions plus a transition-metal oxide framework. However, their atomic and electronic structure evolution during electrochemical cycling remains poorly understood. Here we report the in situ observation of the three-dimensional structural evolution of the transition-metal oxide framework in an all-solid-state battery. The in situ studies LiNi0.5Mn1.5O4 from various zone axes reveal the evolution of both atomic and electronic structures during delithiation, which is found due to the migration of oxygen and transition-metal ions. Ordered to disordered structural transition proceeds along the <100>, <110>, <111> directions and inhomogeneous structural evolution along the <112> direction. Uneven extraction of lithium ions leads to localized migration of transition-metal ions and formation of antiphase boundaries. Dislocations facilitate transition-metal ions migration as well. Theoretical calculations suggest that doping of lower valence-state cations effectively stabilize the structure during delithiation and inhibit the formation of boundaries.


An improved vehicle-pedestrian near-crash identification method with a roadside LiDAR sensor.
Potential conflicts between pedestrians and vehicles represent a challenge to pedestrian safety. Near-crash is used as a surrogate metric for pedestrian safety evaluations when historical vehicle-pedestrian crash data are not available. One challenge of using near-crash data for pedestrian safety evaluation is the identification of near-crash events. This paper introduces a novel method for pedestrian-vehicle near-crash identification that uses a roadside LiDAR sensor. The trajectory of each road user can be extracted from roadside LiDAR data via several data processing algorithms: background filtering, lane identification, object clustering, object classification, and object tracking. Three indicators, namely, the post encroachment time (PET), the proportion of the stopping distance (PSD), and the crash potential index (CPI) are applied for conflict risk classification. The performance of the developed method was evaluated with field-collected data at four sites in Reno, Nevada, United States. The results of case studies demonstrate that pedestrian-vehicle near-crash events could be identified successfully via the proposed method. Practical applications: The proposed method is especially suitable for pedestrian-vehicle near-crash identification at individual sites. The extracted near-crash events can serve as supplementary material to naturalistic driving study (NDS) data for safety evaluation.


The importance of radiographic imaging in the microscopic assessment of bone tumors.
Primary bone tumors are rare and require a multidisciplinary approach. Diagnosis involves primarily the radiologist and the pathologist. Bone lesions are often heterogeneous and the microscopic diagnostic component(s) may be in the minority, especially on core needle biopsies. Reactive processes, benign, and malignant tumors may have similar microscopic aspects. For these challenging cases, the correlation of microscopic and radiologic information is critical, or diagnostic mistakes may be made with severe clinical consequences for the patient. The purpose of this article is to explain how pathologists can best use imaging studies to improve the diagnostic accuracy of bone lesions. Many bone lesions are microscopically and/or radiographically heterogeneous, especially those with both lytic and matrix components. Final diagnosis may require specific microscopic diagnostic features that may be present in the lesion, but not the biopsy specimen. A review of the imaging helps assess if sampling was adequate. The existence of a pre-existing bone lesion, syndrome (such as Ollier disease or multiple hereditary exostosis), or oncologic history may be of crucial importance. Finally, imaging information is very useful for the pathologist to perform accurate local and regional staging during gross examination. Close teamwork between pathologists, radiologists, and clinicians is of utmost importance in the evaluation and management of bone tumors. These lesions can be very difficult to interpret microscopically; imaging studies therefore play a crucial role in their accurate diagnosis.


Fusion Imaging and Virtual Navigation to Guide Percutaneous Thermal Ablation of Hepatocellular Carcinoma: A Review of the Literature.
As medical imaging advancements have improved the detectability of hepatocellular carcinoma (HCC) in early stages, the approach to percutaneous thermal ablation for curative treatment has concomitantly advanced. Although many centers are adopting cross-sectional imaging to guide percutaneous ablation, the majority of procedures are still performed under ultrasound (US) guidance worldwide. Challenges to ultrasound guidance may present due to relatively poor resolution particularly with small or isoechoic lesions, or due to intervening structures such as the bowel or diaphragm that obstruct lesional visualization. Fusion imaging (FI) systems have been employed to address these challenges. By merging or synchronizing the real-time images from US with a previously obtained cross-sectional study, FI mitigates the inherent limitations of each individual imaging modality and expands procedural feasibility and technical outcomes. This manuscript reviews the current literature on the use of FI during percutaneous thermal ablation of HCC.


Postsynaptic endocannabinoid release is critical to long-term depression in the striatum.
The striatum functions critically in movement control and habit formation. The development and function of cortical input to the striatum are thought to be regulated by activity-dependent plasticity of corticostriatal glutamatergic synapses. Here we show that the induction of a form of striatal synaptic plasticity, long-term depression (LTD), is dependent on activation of the CB1 cannabinoid receptor. LTD was facilitated by blocking cellular endocannabinoid uptake, and postsynaptic loading of anandamide (AEA) produced presynaptic depression. The endocannabinoid necessary for striatal LTD is thus likely to be released postsynaptically as a retrograde messenger. These findings demonstrate a new role for endocannabinoids in the induction of long-term synaptic plasticity in a circuit necessary for habit formation and motor control.


Inhibition of GLUT2 transporter by geraniol from Cymbopogon martinii: a novel treatment for diabetes mellitus in streptozotocin-induced diabetic rats.
To isolate and identify the bioactive component from Cymbopogon martinii having GLUT2 transporter inhibitory activity - towards development of a novel strategy for treatment of diabetes mellitus. Isolation of bioactive component was carried out using differential solvent extraction, HPTLC and HPLC, and identification was done by GC-MS. In-vitro studies on intestine, liver, kidney and in-vivo assessment by OGTT and long-term treatment on diabetic rats were carried out. Geraniol was isolated and identified as bioactive component. Intestinal glucose absorption demonstrated 60.28% inhibition of transport at 648.34 μm of geraniol. It was found to inhibit glucose release from liver on adrenaline challenge by 89.82% at 324.17 μm/ml. Kidney glycogen content doubled using 648.34 μm of geraniol as compared to control. Geraniol demonstrated 2.14 times higher renal glucose output than diabetic control. OGTT demonstrated prevention of postprandial spikes. Prolonged treatment for 60 days with 29.37 mm/kg B.W. twice a day of geraniol improved the lipid profile, HbA1C levels and renal parameters. In mRNA studies for 10 days, over expression of GLUT2 was prevented by geraniol. Inhibition of GLUT2 by geraniol has the potential to reduce hyperglycaemia and prevent secondary complications in diabetes.


Pulsed nuclear magnetic resonance study of 39K within halobacteria.
The 39K contents of isolated pellets and supernatant solutions from suspensions of Halobacterium halobium were studied at 21-22 degrees C by pulsed NMR spectroscopy. The rates of transverse relaxation were measured directly from the free induction decay (FID). The rate of longitudinal relaxation was measured by studying the FID after pairs of pulses of approximately 90 degrees. Care was exercised to minimize the effect of magnetic field inhomogeneity; its contribution to the FID was approximately 25-30 sec-1. The transverse relaxation process was found to consist of at least two components, whose rates were 321-449 sec-1 and 1,122-2,067 sec-1. In one preparation where the longitudinal relaxation process was studied, the data could be well fit to a single exponential relaxing at 253 +/- 33 (mean +/-95% confidence limits) sec-1. Comparison of the relative intensities of the NMR signals with the results of atomic absorption photometric analyses indicated that the great bulk of the intracellular 39K was detected by the NMR techniques used. The data obtained from the current NMR of H. halobium are consistent with: (1) fractional binding of less than 3% of the total intracellular K+, (2) a small ordering factor characterizing all of the intracellular K+, or (3) some combination of the two.


DNA demethylase activity maintains intestinal cells in an undifferentiated state following loss of APC.
Although genome-wide hypomethylation is a hallmark of many cancers, roles for active DNA demethylation during tumorigenesis are unknown. Here, loss of the APC tumor suppressor gene causes upregulation of a DNA demethylase system and the concomitant hypomethylation of key intestinal cell fating genes. Notably, this hypomethylation maintained zebrafish intestinal cells in an undifferentiated state that was released upon knockdown of demethylase components. Mechanistically, the demethylase genes are directly activated by Pou5f1 and Cebpβ and are indirectly repressed by retinoic acid, which antagonizes Pou5f1 and Cebpβ. Apc mutants lack retinoic acid as a result of the transcriptional repression of retinol dehydrogenase l1 via a complex that includes Lef1, Groucho2, Ctbp1, Lsd1, and Corest. Our findings imply a model wherein APC controls intestinal cell fating through a switch in DNA methylation dynamics. Wild-type APC and retinoic acid downregulate demethylase components, thereby promoting DNA methylation of key genes and helping progenitors commit to differentiation.


Extracellular Signal-Regulated Kinase1 (ERK1)-Mediated Phosphorylation of Voltage-Dependent Anion Channel (VDAC) Suppresses its Conductance.
ERK1 is one of the members of the mitogen-activated protein kinases that regulate important cellular functions. VDAC is located at the outer membrane of mitochondria. Here, an interaction between VDAC and ERK1 has been studied on an artificial planar lipid bilayer using in vitro electrophysiology experiments. We report that VDAC is phosphorylated by ERK1 in the presence of Mg2+-ATP and its single-channel currents are inhibited on the artificial bilayer membrane. Treatment of Alkaline phosphatase on ERK1 phosphorylated VDAC leads to partial recovery of the single-channel VDAC currents. Later, phosphorylation of VDAC was demonstrated by Pro-Q diamond dye. Mass Spectrometric studies indicate phosphorylation of VDAC at Threonine 33, Threonine 55, and Serine 35. In a nutshell, phosphorylation of VDAC leads to the closure of the channel.


Treatment of acanthosis nigricans with sequential salicylic acid-mandelic acid combination peel and maintenance with glycolic acid-urea combination cream: A retrospective pilot study.
Acanthosis nigricans (AN) is a common dermatosis that presents with hyperpigmented, velvety thick plaques over intertriginous areas. Though a number of treatment modalities including chemical peels have been used, none provide long-term and sustained improvement. Our study evaluated the efficacy and safety of regular sessions of salicylic acid-mandelic acid peeling over axillary AN lesions, which was followed by daily application of a topical combination of glycolic acid, urea, and cetylated fat esters for maintenance of effect for 9 months. A retrospective pilot study was conducted in Indian patients (Fitzpatrick skin type 4 or 5), aged 18-50 with benign hereditary AN involving the underarms, with or without affection of other typical sites. Data were retrieved of seventeen patients with AN involving the axillae. Patients were started on combination salicylic-mandelic acid peel given every 2 weeks for a total of 6 sessions. Maintenance was done by night application of combination cream of glycolic acid, urea, and cetylated fat esters, which was continued for 9 months after completion of peeling sessions. Lesions were evaluated every 3 months of 9 months for improvement in pigmentation and skin thickening. All the patients (100%) showed significant improvement in both pigmentation and thickening of lesions. In terms of improvement in skin thickening, very good improvement was seen in 41%, while 29% patients had moderate improvement. In terms of improvement in pigmentation, 35% each had very good and moderate improvement. Post peel erythema (100%) and burning sensation (90%) were the most common encountered adverse effects which lasted for only 1-2 days. Patients were followed up for another 9 months during which no relapses were seen. Combination of keratolytic chemical peels and topical mild keratolytic application ensures better therapeutic outcome in patients of AN with long lasting effect.


Variance in relative internode length (l/d) in the rat and its presumed significance for the safety factor and neuropathy.
(1) Previous work has shown that the quotient l/d (internode length/fiber diameter) is the product of independent variations in internode length (increasing with fiber elongation) and in axon caliber (whose growth is not as yet well understood). Using these guiding principles, the distribution of l/d in 19 nerves and 7 roots of the rat was determined. (2) Each nerve or root showed a linear decrease of the l/d with fiber caliber, the thin fibers always having relatively longer internodes than the thick ones. Comparing nerves, the highest l/d was found for extremity nerves, particularly those in the hindlimbs. Nerves of the trunk (phrenic, intercostal) had lower l/d with further decreases for the extracranical branches of cranical nerves, the intercranial roots and a minimum for the acoustic nerve. In the facial nerve, the l/d of the intraosseous portions was distinctly lower than that of its branches in the face. Ventral roots showed a cranio-caudal increase in their l/d. For some fiber systems, e.g. cranial nerves, the l/d reflected variance in axon caliber, their internode length being relatively uniform. For others, e.g. roots, the l/d was dominated by their large variation in internode length. (3) Variation of the l/d in fiber populations may correlate with such parameters as internodal conduction time, different sensitivities of thick and thin fibers and, particularly, the safety factor. It is also proposed that the regional variation in the l/d is a critical parameter affecting the regional vulnerability of fiber populations to polyneuropathies or to radiculopathies.


Human leukocyte antigen-G 14 bp insertion/deletion polymorphism contributes to preeclampsia risk in Asian population: A systematic review and meta‑analysis.
Preeclampsia remains enigmatic and responsible for vast maternal and fetal morbidity and mortality worldwide. Our objective was to assess the strength of the effect of the 14 bp deletion/insertion polymorphism in exon 8 of the 3'UTR region of the human leukocyte antigen-G (HLA-G) gene on preeclampsia risk across different populations. A systematic review by a meta-analysis was performed to summarize the scattered epidemiologic evidence, which remains inconclusive and controversial. A systematic literature search according to the PRISMA guidelines was conducted to screen relevant publications. Odds ratio and corresponding 95% confidence interval were estimated to measure the magnitude of the association between this polymorphism and preeclampsia onset. Thirty studies comprising 9402 subjects were eligible. Pooled estimates suggested that both fetal and paternal insertion variants were significantly associated with increased odds of this disease. Nevertheless, the presence of the 14 bp insertion sequence in mothers does not seem to increase the risk of preeclampsia. Moreover, the results of subgroup analysis suggested that the fetal, maternal, and paternal polymorphism has a significant deleterious impact on the preeclampsia risk in the Asian population. In addition, the significant association between the paternal polymorphism and preeclampsia in primigravida was observed in the pooled estimation with a small sample size. By summarizing the amount of significant evidence, our study nominated this polymorphism as a potential biomarker for early risk stratification for Asians. Further large-scale validation is needed to establish fully solid and conclusive evidence for the impact of the insertion polymorphism on preeclampsia risk.


Knowledge is power: studying critical incidents in intensive care.
Despite their difficult definition and taxonomy, it is imperative to study critical incidents in intensive care, since they may be followed by adverse events and compromised patient safety. Identifying recurring patterns and factors contributing to critical incidents constitutes a prerequisite for developing effective preventive strategies. Self-reporting methodology, although widely used for studying critical incidents, has been criticized in terms of reliability and may considerably underestimate both overall frequency and specific types of them. Promotion of non-blaming culture, analysis of critical incident reports and development of clinical recommendations are expected to minimize critical incidents in the future.


Isolation of two immunologically related transglutaminase substrates from cultured human keratinocytes.
Transglutaminase substrates A and B were identified in soluble extracts of cultured keratinocytes and human epidermis by their reactions with dansyl cadaverine in the presence of Ca++ ion. Substrate B was present in substantial amounts in both extracts whereas A, easily seen in cell extracts, was decreased and sometimes not detected in tissue extracts. Substrates A and B from cultured cells were separated by Sephadex G-75 chromatography and isolated by preparative sodium dodecyl sulfate (SDS) gel electrophoresis by which A had a mol wt of 125,000 and B had a mol wt of 12,000. Amino acid analysis of A, B, and cornified envelope were similar but not identical. The isopeptide bond is not a significant structural feature of A inasmuch as its content is less than 0.25 bonds/molecule. Antibodies raised to A cross-reacted with B and vice versa and A showed partial identity to B when reacted with anti-B. Anti-A reacted with epidermis being adsorbed by the edges of cornifying cells but only weakly by cells of the Malpighian layer. Anti-B also reacted with cornifying, but its reaction was more intense with the cytoplasm of Malpighian cells. Substrate A appears to be incorporated into cornified envelope immediately after its appearance in cells of the granular layer and seems similar to a protein isolated by a different method. Substrate B, convertible by transglutaminase to higher molecular weight species, may also participate in cornified envelope assembly and shares some structural similarities to A.


Does the Microbiology of Bactibilia Drive Postoperative Complications After Pancreatoduodenectomy?
The influence of bile microflora, particularly with broad antimicrobial resistance patterns, on postoperative outcomes after pancreatoduodenectomy (PD), is poorly understood. The aim of this study was to determine the influence of the microbiology of bactibilia on postoperative outcomes following PD. Intraoperative bile cultures were obtained in 162 patients undergoing PD between 2015 and 2017. Intraoperative bile cultures were analyzed and correlated with short-term outcomes after PD. Independent groups t test, Pearson's correlation, or Fisher's exact tests were performed. Hazard ratios (HR) are reported with 95% confidence intervals (CI). Statistical significance was defined as P value of < 0.05. Intraoperative bile cultures were positive in 89/162 patients (55%). The most common bacteria were Enterococcus spp. (n = 48, 54%), Klebsiella spp. (n = 24, 27%), and Enterobacter spp. (n = 17, 19%). Bactibilia was not associated with increased infectious complications, postoperative pancreatic fistula (POPF), or mortality. Enterococcus and Enterobacter were associated with higher rates of incisional (HR, 6.5; 95% CI, 1.2-34.8; P = 0.03) and organ-space surgical site infection (HR, 4.9; 95% CI, 1.1-22.0; P = 0.03), respectively. No single bacterium was associated with POPF, bile leak, cholangitis, 30- or 90-day mortality. Bactibilia, in general, does not increase the risk of developing a postoperative complication following pancreatoduodenectomy; however, Enterococcus and Enterobacter increase the likelihood of developing incisional and organ-space surgical infections, respectively.


The effect of epithelium removal on non-adrenergic, non-cholinergic inhibitory responses in the isolated central airways of the cat and guinea pig.
Epithelium removal from the feline or the indomethacin-treated guinea pig trachea had no effect on tissue sensitivity or responsiveness to the contractile actions of pharmacological agonists or electrical field stimulation. In the feline hilar bronchus, epithelium removal had no effect on tissue sensitivity or responsiveness to acetylcholine or electrical field stimulation but increased bronchial sensitivity to serotonin without affecting responsiveness. Non-adrenergic non-cholinergic (NANC) relaxation responses elicited by electrical field stimulation in airway preparations from either species were unaffected by epithelium removal. These results suggest that the epithelium does not modulate contractile responses in the feline trachea but may modulate the actions of specific contractile agonists in the feline hilar bronchus. Further, NANC relaxation responses appear to occur independently of the airway epithelium.