"Zero reference" during cardiac catheterization.
Hemodynamic pressure measurements obtained during cardiac catheterization studies are calibrated to an arbitrary level where the pressure is considered zero. Despite the fundamental nature of the zero reference level, there is no standardized method for determination of "zero reference." A horizontal metal rod has been constructed to be level with the pressure transducer domes. With lateral radiographic viewing, the metal rod is adjusted to be level with the junction of the inferior vena cava and the right atrium. The technique is rapid, reliable, without risk and allows accurate comparison of hemodynamic data between patients and in the same patient longitudinally.


Increased expression of the tight junction protein TJP1/ZO-1 is associated with upregulation of TAZ-TEAD activity and an adult tissue stem cell signature in carfilzomib-resistant multiple myeloma cells and high-risk multiple myeloma patients.
Tight junction protein 1 (TJP1) has recently been proposed as a biomarker to identify multiple myeloma (MM) patients most likely to respond to bortezomib- and carfilzomib-based proteasome inhibitor regimens. Herein we report increased expression of TJP1 during the adaptive response mediating carfilzomib resistance in the LP-1/Cfz MM cell line. Moreover, increased TJP1 expression delineated a subset of relapsed/refractory MM patients on bortezomib-based therapy sharing an LP-1/Cfz-like phenotype characterized by activation of interacting transcriptional effectors of the Hippo signaling cascade (TAZ and TEAD1) and an adult tissue stem cell signature. siRNA-mediated knockdown of TJP1 or TAZ/TEAD1 partially sensitized LP-1/Cfz cells to carfilzomib. Connectivity Map analysis identified translation inhibitors as candidate therapeutic agents targeting this molecular phenotype. We confirmed this prediction by showing that homoharringtonine (omacetaxine mepesuccinate) - the first translation inhibitor to be approved by the U.S. Food and Drug Administration - displayed potent cytotoxic activity on LP-1/Cfz cells. Homoharringtonine treatment reduced the levels of TAZ and TEAD1 as well as the MM-protective proteins Nrf2 and MCL1. Thus, our data suggest the importance of further studies evaluating translation inhibitors in relapsed/refractory MM. On the other hand, use of TJP1 as a MM biomarker for proteasome inhibitor sensitivity requires careful consideration.


Myelinoclastic diffuse sclerosis (Schilder's disease) is immunologically distinct from multiple sclerosis: results from retrospective analysis of 92 lumbar punctures.
Myelinoclastic diffuse sclerosis (MDS; also termed Schilder's disease) is a rare inflammatory demyelinating disorder of the central nervous system characterised by demyelination of vast areas of the white matter. It is unclear whether MDS is a variant of multiple sclerosis (MS) or a disease entity in its own right. To compare the cerebrospinal fluid (CSF) features of MDS with those of MS. Retrospective analysis of the CSF profile of all patients with MDS reported in the medical literature between 1960 and 2018. The most striking finding was a substantial lack of oligoclonal bands (OCBs) in MDS, which were absent in at least 77% (30/39) of all lumbar punctures (LP) in the total cohort and in 86% in the subgroup of patients with normal very long-chain fatty acid serum ratios (VLCFA). Almost all cases published in the past 15 years were negative for OCBs. These findings are in contrast to MS, in which OCBs are present in up to 98% of cases (p < 0.00001 when compared with reference works in MS; both in adult and in pediatric patients). CSF pleocytosis was absent in at least 79% (46/58) of all LP (p < 0.0001 vs. MS) and in 92% (24/26) of LPs in the VLCFA-tested subgroup. CSF total protein levels were elevated in 56% of all LPs (p < 0.0001 vs. MS) and in 63% of LPs in the VLCFA-tested subgroup and were often higher than in typical MS (> 100 mg/dL in 13/22; up to 220 mg/dL). EBV serum antibodies, which are present in virtually all patients with MS, and the so-called MRZ (measles/rubella/zoster) reaction, a highly specific marker of MS, were absent in all of the few patients tested. In addition, we discuss further differences between MS and MDS, taking into account also Schilder's original comprehensive case description from 1912. In the majority of patients diagnosed with MDS, CSF features differ significantly from those typically found in MS and are more similar to those previously reported in patients with myelin oligodendrocyte glycoprotein-immunoglobulin G (IgG)-positive encephalomyelitis, aquaporin-4-IgG-positive neuromyelitis optica spectrum disorders or Baló's concentric sclerosis. Our data suggest that MDS and MS are immunopathologically distinct entities in the majority of cases.


Identification and characterization of a heme exporter from the MRP family in Drosophila melanogaster.
The heme group constitutes a major functional form of iron, which plays vital roles in various biological processes including oxygen transport and mitochondrial respiration. Heme is an essential nutrient, but its pro-oxidant nature may have toxic cellular effects if present at high levels, and its synthesis is therefore tightly regulated. Deficiency and excess of heme both lead to pathological processes; however, our current understanding of metazoan heme transport is largely limited to work in mammals and the worm Caenorhabditis elegans, while functional analyses of heme transport in the genetically amenable Drosophila melanogaster and other arthropods have not been explored. We implemented a functional screening in Schneider 2 (S2) cells to identify putative heme transporters of D. melanogaster. A few multidrug resistance-associated protein (MRP) members were found to be induced by hemin and/or involved in heme export. Between the two plasma membrane-resident heme exporters CG4562 and CG7627, the former is responsible for heme transit across the intestinal epithelium. CG4562 knockdown resulted in heme accumulation in the intestine and lethality that could be alleviated by heme synthesis inhibition, human MRP5 (hMRP5) expression, heme oxygenase (HO) expression, or zinc supplement. CG4562 is mainly expressed in the gastric caeca and the anterior part of the midgut, suggesting this is the major site of heme absorption. It thus appears that CG4562 is the functional counterpart of mammalian MRP5. Mutation analyses in the transmembrane and nucleotide binding domains of CG4562 characterized some potential binding sites and conservative ATP binding pockets for the heme transport process. Furthermore, some homologs in Aedes aegypti, including that of CG4562, have also been characterized as heme exporters. Together, our findings suggest a conserved heme homeostasis mechanism within insects, and between insects and mammals. We propose the fly model may be a good complement to the existing platforms of heme studies.


Solid-state NMR characterization of gas vesicle structure.
Gas vesicles are gas-filled buoyancy organelles with walls that consist almost exclusively of gas vesicle protein A (GvpA). Intact, collapsed gas vesicles from the cyanobacterium Anabaena flos-aquae were studied by solid-state NMR spectroscopy, and most of the GvpA sequence was assigned. Chemical shift analysis indicates a coil-α-β-β-α-coil peptide backbone, consistent with secondary-structure-prediction algorithms, and complementary information about mobility and solvent exposure yields a picture of the overall topology of the vesicle subunit that is consistent with its role in stabilizing an air-water interface.


Follicle development during the normal menstrual cycle.
An understanding of the factors which determine initiation of follicle growth, recruitment and dominant follicle selection may increase our understanding of the underlying process of ovarian aging. In this article, these aspects of the normal menstrual cycle are reviewed. The morphological and endocrinological development in the early follicle is described from the primordial follicle stage. The degree of follicle-stimulating hormone (FSH) dependency is discussed, as is the relationship of estradiol (E2) production to follicle diameter. The principles governing mono-follicular selection are outlined, and the FSH 'threshold' and 'window' concepts are highlighted. Maximum FSH levels in the early follicular phase are shown to be variable between individuals. The relevance of this and the means by which individual sensitivity to FSH may be altered at the ovary in the context of ovarian aging are discussed.


Proton-induced sodium current in freshly dissociated hypothalamic neurones of the rat.
1. The proton-gated current was investigated in freshly dissociated ventromedial hypothalamic (VMH) neurones from 4-week-old Wistar rats, under whole-cell configuration by the use of the 'concentration-clamp' technique which combines intracellular perfusion with the rapid exchange of external solution within 1-2 ms under a single-electrode voltage-clamp condition. 2. The proton-gated current increased in a sigmoidal fashion as extracellular pH (pHo) decreased. In external solution containing 2 mM-Ca2+, the threshold of current activation was at pHo 6.5, and the maximum response appeared at pHo 4.1-3.9. The dissociation constant (Kd) and Hill coefficient were 10(-4.9) M (pHo = 4.9) and 1.5 respectively. 3. Decreasing extracellular Na+ concentration reduced the proton-gated current. The current reversed direction at the Na+ equilibrium potential (ENa), indicating that it was carried by Na+. 4. The activation phase kinetics of proton-induced current was single exponential. The time constant of activation (tau a) did not have a potential dependence but decreased slightly by decreasing pHo. The inactivation phase kinetics was two-exponential. The time constant of inactivation (tau i) consisted of fast and slow components (tau if and tau is, respectively). Like tau a, both tau if and tau is did not have any potential dependence, but they slightly increased with decreasing pHo. 5. The steady-state inactivation curve, constructed by decreasing pHo from various conditioning pHos to 4.1, revealed that the proton-induced current had a half-maximum inactivation at pHo 6.2. 6. The proton-induced current was suppressed as the extracellular Ca2+ concentration ([Ca2+]o) increased from almost free (0.01 mM) to 80 mM. Increasing [Ca2+]o increased tau a, but slightly decreased both tau if and tau is. 7. Recovery of proton-induced current from complete inactivation of proton-induced current depended on the degree of pHo change. A bigger change in pHo induced faster recovery than a smaller change. 8. External divalent cations inhibited the proton-induced current, and the inhibitory potency was in the order of Mn2+ greater than Co2+ greater than Ca2+ greater than Sr2+ greater than Ba2+ greater than Mg2+. 9. Tetrodotoxin (TTX) at relatively low concentration (less than 10(-7) M) did not inhibit the peak amplitude of the proton-induced current, but at a higher concentration (10(-6) M) it slightly inhibited the peak amplitude of the current and accelerated the inactivation process. Scorpion toxin markedly increased the peak amplitude of the proton-induced current and prolonged the inactivation phase. The tau is was also increased by scorpion toxin in a concentration-dependent manner. Veratridine had no effect on the proton-induced current. 10. The membrane properties of the proton-operated channel were similar to those of the voltage-gated Na+ channel rather than the Ca2+ channel.


[Hip septic arthritis in young children with sickle-cell disease].
Sickle-cell anemia is an inherited disorder exposing patients to diverse complications, particularly bone and joint disease. Septic arthritis of the hip joint is a frequent infection in sickle-cell anemia. Without early management, there is an important risk of invalidating hip dysfunction. The purpose of this study was to analyze the epidemiological, clinical and therapeutic aspects of septic arthritis of the hip joint in order to focus on the need for early diagnosis, the best way to improve the prognosis of hip function. This was a retrospective study over a 10 year period (1987-1996) which included nine children (seven boys and two girls), mean age six years who were managed in our hospital for septic arthritis of the hip joint. These children had at least one hemoglobin S: phenotypes SS (n=5), SC (n=2), AS (n=2). In these sickle-cell anemia children, the septic arthritis developed in a non traumatic context with osteonecrosis of the femoral head. One hip was involved in six children and both in three (12 hips). Mean time from symptom onset to consultation was eight days. Salmonella sp. were identified in nine hips (including all of the bilateral cases) and Staphylococcus sp. in three. The treatment included antibiotics for six weeks, puncture-drainage, and immobilization for three to four weeks. Outcome was excellent in nine hips and good in three. The diagnosis of septic arthritis of the hip joint must be made early, based on ultrasound or scintigraphic findings. Bacteriological samples of joint fluids are required to adjust the antibiotic regimen. Salmonella sp. were the most common agents in our patients, but other series have reported Klebsiella pneumoniae and Staphylococcus or Pneumococcus sp. Drainage is preferred, if possible by arthroscopy. Arthrotomy should be reserved for cases with a thick discharge which is difficult to evacuate via puncture drainage. Early diagnosis, careful evacuation of the joint collection, and adapted antibiotic therapy with hip immobilization are the keys to good functional outcome.


Decreasing Barriers to Research Utilization Among Labor and Delivery Nurses.
Intermittent fetal monitoring (IFM) is a recommended strategy for intrapartum fetal heart rate assessment in low-risk pregnancies; however, this "high touch, low tech" approach is underutilized. The aim of the study was to examine the relationships between labor and delivery nurses' intellectual capital and their perceptions of barriers to research utilization in the work setting. A cross-sectional correlational design using data derived from a larger study of labor and delivery nurses (N = 248) was used. Covell's theory of nursing intellectual capital was used as the conceptual and analytic framework to examine labor and delivery nurses' intellectual capital and their perceived barriers to research utilization. Nurses who receive paid time off from their employer to attend conferences (p < .01) and nurses who do not report nurse-to-patient ratios as a problem in providing IFM (p < .01) perceive fewer barriers to research utilization. Time, especially available time, has an effect on labor and delivery nurses' attitude toward IFM and their perceptions of barriers to research utilization.


Neuromuscular complications of intensive care.
Critical illness polyneuropathy and critical illness myopathy are well recognized in adults, in whom they commonly cause generalized weakness and muscle wasting, with failure to wean from mechanical ventilation. There is significant clinical and neurophysiological overlap between the two conditions. Critical illness polyneuropathy and critical illness myopathy can cause significant morbidity in critically ill children, but they have only occasionally been reported in childhood, and little is known of their prevalence or clinical significance in this population. These conditions seem to be clinically and electrophysiologically similar in children and adults, but prospective studies of these entities are required to better characterize their frequency, natural history, and clinical significance in pediatric practice.


Coronary clearance frame count: a new index of microvascular perfusion.
The angiographic data of myocardial perfusion obtained following direct angioplasty in acute myocardial infarction (AMI) can be evaluated only by qualitative methods: the myocardial blush grade (MBG) or the TIMI myocardial perfusion grade (TMPG). To minimize the subjective nature of this evaluation, we describe a quantitative index, the "coronary clearance frame count" (CCFC); and have tested it against known indices. All patients with AMI treated with direct angioplasty in a single center over 2 years were prospectively analyzed. All films were assessed off-line to determine the traditional indices of TIMI flow grade, TIMI frame count, MBG and TMPG. To define CCFC, we used the first frame in which the contrast is cleared from the ostium of the artery as "frame 0" and that in which contrast begins to wash-in into the same coronary artery distal landmark proposed by TIMI Group, as the "last frame". Of the 147 patients studied, 110 had films that were technically adequate for measuring qualitative and quantitative indices of myocardial perfusion. CCFC showed a good correlation with MBG (p=0.045) and with TMPG (p<0.001). CCFC was strongly related to the presence of TMPG 2 or 3 (p<0.001). A cut-off value of 45 frames has a sensitivity of 75% and specificity of 70% to predict a TMPG 2 or 3. Coronary clearance frame count has a good correlation with known indices of reperfusion and has the advantage of being an objective, quantitative index that is efficient even in inexperienced hands. Abbreviated Abstract. The angiographic quality of myocardial perfusion data obtained following direct angioplasty in acute myocardial infarction can only be evaluated by qualitative methods: the myocardial blush grade or the TIMI myocardial perfusion grade. To minimize the subjective nature of these methods, we describe a quantitative index (the "coronary clearance frame count") which we evaluated against the other well-established indices, and have observed it to be efficient even in the hands of inexperienced practitioners.


Treatment Adherence in CKD and Support From Health care Providers: A Qualitative Study.
Adherence to recommended medical treatment is critical in chronic kidney disease (CKD) to prevent complications and progression to kidney failure. Overall adherence to treatment is low in CKD, and as few as 40% of patients with kidney failure receive any documented CKD-related care. The purpose of this study was to explore the experiences of patients with CKD and their adherence to CKD treatment plans, and the role their health care providers played in supporting their adherence. One-on-one interviews were conducted in 2019-2020 using a semi-structured interview guide. Participants described experiences with adherence to treatment plans and what they did when experiencing difficulty. Participants were recruited from the Chronic Renal Insufficiency Cohort (CRIC) study. All CRIC participants were older than 21 years with CKD stages 2-4; this sample consisted of participants from the University of Pennsylvania CRIC site. Interviews were recorded, transcribed, and coded using conventional content analysis. Data were organized into themes using NVivo 12. The sample (n = 32) had a mean age of 67 years, 53% were women, 59% were non-White, with a mean estimated glomerular filtration rate of 56.6 mL/min/1.73 m2. From analysis of factors relevant to treatment planning and adherence, following 4 major themes emerged: patient factors (multiple chronic conditions, motivation, outlook), provider factors (attentiveness, availability/accessibility, communication), treatment planning factors (lack of plan, proactive research, provider-focused treatment goals, and shared decision making), and treatment plan responses (disagreeing with treatment, perceived capability deficit, lack of information, and positive feedback). The sample was drawn from the CRIC study, which may not be representative of the general population with CKD. These themes align with Behavioral Learning Theory, which includes concepts of internal antecedents (patient factors), external antecedents (provider factors), behavior (treatment planning factors), and consequences (treatment plan responses). In particular, the treatment plan responses point to innovative potential intervention approaches to support treatment adherence in CKD.


Long-term follow-up outcomes for patients undergoing primary total hip arthroplasty with uncemented versus cemented femoral components: a retrospective observational study with a 5-year minimum follow-up.
This retrospective analysis compared the long-term outcomes for patients with a femoral neck fracture (AO/OTA type 31B) treated with a primary unilateral total hip arthroplasty with uncemented or cemented femoral components (UTHA or CTHA, respectively). We conducted a retrospective cohort study using the South China Hip Arthroplasty Database. We identified 422 patients with femoral neck fracture (AO/OTA type 31B) who were previously treated with primary unilateral UTHA or CTHA between 2007 and 2015, with follow-up until 2019. Follow-up occurred 1, 3, 6 and 12 months postoperatively and yearly thereafter. The primary outcome was the Harris hip score (HHS). The secondary outcome was the orthopaedic complication rate. In total, 324 patients (UTHA n = 160, mean age 68.61 ± 7.49 years; CTHA n = 164, mean age 68.75 ± 7.04 years) were evaluated for study eligibility. The median follow-up was 73.3 months (range, 11.6-89.2 months). At the final follow-up, HHS was 74.09 ± 6.23 vs 79.01 ± 10.21 (UTHA vs CTHA, p = 0.012). Significant differences were detected in the incidence of prosthetic revision, loosening, and periprosthetic fracture between the UTHA and CTHA groups (7.5% for UTHA vs 1.8% for CTHA, p = 0.015; 17.5% for UTHA vs 8.5% for CTHA, p = 0.016; 11.9% for UTHA vs 4.9% for CTHA, p = 0.021, respectively). In this setting, CTHA demonstrated superiority to UTHA by improving functional outcomes and decreasing complication rates.


Medical Therapy and Coronary Revascularization for Patients With Stable Coronary Artery Disease and Unclassified Appropriateness Score.
Although the appropriate use criteria incorporate common clinical scenarios for coronary revascularization, a significant proportion of patients with stable coronary artery disease (CAD) cannot be assigned an appropriateness score. Our objective was to characterize these patients and to evaluate whether coronary revascularization is associated with improved outcomes. A population-based cohort of patients aged ≥66 years, who underwent cardiac catheterization in Ontario, Canada, were included. Clinical characteristics were compared between patients with and without an appropriateness score. Clinical outcomes between coronary revascularization and medical therapy in patients with unclassified appropriateness score were compared using the inverse probability of treatment-weighted propensity method for confounder adjustment. Of the 19,228 patients with stable CAD, 11.2% (2,153 patients) were not assigned to an appropriateness score, mostly (92.9%) because of a lack of ischemic evaluation or a noninterpretable test. These patients were older, had higher rate of severe angina, and had more medical co-morbidities compared to patients with an appropriateness score. The 2-year rate of death or myocardial infarction in patients with unclassified appropriateness score was 15.3% in the revascularization group versus 20.7% in the medical therapy group. After propensity weighting, revascularization was associated with significantly lower hazard ratio (0.70; 95% confidence interval 0.61 to 0.79) for death or myocardial infarction compared with medical therapy. In conclusion, in patients aged ≥66 years with stable CAD and unclassified appropriateness score, revascularization is associated with improved outcomes.


Muscle fiber typology is associated with the incidence of overreaching in response to overload training.
The aim of this study was to identify markers of training stress and characteristics of middle-distance runners related to the incidence of overreaching following overload training. Twenty-four highly trained middle-distance runners [n = 16 male, peak oxygen uptake (V̇o2peak) = 73.3 (4.3) mL·kg·min-1; n = 8 female, V̇o2peak = 63.2 (3.4) mL·kg·min-1] completed 3 wk of normal training (NormTr), 3 wk of high-volume training (HVTr; a 10%, 20%, and 30% increase in training volume each successive week from NormTr), and 1 wk of taper (TapTr; 55% exponential reduction in training volume from HVTr week 3). Before and immediately after each training period, an incremental treadmill-running test was performed, while resting metabolic rate (RMR), subjective fatigue responses, and various resting blood biomarkers were assessed. Muscle fiber typology of the gastrocnemius was estimated by quantification of muscle carnosine using proton magnetic resonance spectroscopy and expressed as a z-score relative to a nonathlete control group. Twelve runners were classified as functionally overreached (FOR) following HVTr [decreased running time to exhaustion (TTE)], whereas the other 12 were classified as acutely fatigued (AF; no decrease in running TTE). The FOR group did not demonstrate systematic alterations in RMR, resting blood biomarkers, or submaximal exercise responses, compared with the AF group. The gastrocnemius carnosine z-score was significantly higher in the FOR group (-0.44 ± 0.57) than in the AF group (-1.25 ± 0.49, P = 0.004, d = 1.53) and was also negatively correlated with changes in running TTE from pre- to post-HVTr (r = -0.55, P = 0.005) and from pre-HVTr to post-TapTr (r = -0.64, P = 0.008). Muscle fiber typology is related to the incidence of overreaching and performance supercompensation following increased training volume and a taper.NEW & NOTEWORTHY Variability in the performance responses following an overload training period and subsequent taper was associated with the variation in the muscle fiber typology of the gastrocnemius. Runners with an estimated higher proportion of type I fibers (i.e., lower carnosine z-score) were able to maintain performance in response to an overload training period and subsequently achieve a superior performance supercompensation. These findings show that muscle fiber typology contributes to the variability in performance responses following training.


Ultra high resolution fMRI at ultra-high field.
In this short review article I will summarize the path we took over the years towards increasing the spatial resolution of fMRI. To fully capitalize on the fMRI technique, a better understanding of the origin of the hemodynamic signals, and what factors are governing their spatial control is necessary. Here, I will briefly describe the studies and developments that ultimately led to our successful effort in mapping orientation columns in humans that is considered by many as the current state-of-the-art for fMRI studies.


Late isolated left ventricular tamponade. Clinical, hemodynamic, and echocardiographic manifestations of a previously unreported postoperative complication.
A unique case of late postoperative left ventricular tamponade, hemodynamically indistinguishable from severe left ventricular failure, is reported in a patient 1 month after an apparently uncomplicated aortic valve replacement. An echo-free space behind the left ventricle led to surgical exploration and successful resolution of the disease process. To our knowledge, this is the first documented case of isolated left ventricular tamponade in the literature. The findings highlight the importance of postoperative echocardiography in the evaluation of the patient having cardiac surgery.


Exon 4-skipping GLT-1: a new form of an abundantly expressed glutamate transporter.
GLT-1 (EAAT2) is an abundant glial glutamate transporter in the mammalian brain. It plays important roles, especially in the termination of neurotransmitter signals at excitatory synapses in grey matter. In normal brain, alternative splicing of GLT-1 has been described, where exons in the GLT-1 gene are skipped or intronic sequences spliced in to generate new sequences. This study describes the isolation of a cDNA clone encoding a new splice variant of GLT-1 where exon 4 is skipped. This novel variant was isolated by RT-PCR cloning from adult rat brain and encodes a protein of 500 amino acids (MW ~54.5 kDa). RT-PCR analysis showed that mRNA was readily detectable in various brain regions of rat, primary astrocyte cultures and in tissues such as testis, but little mRNA was detectable in retina and liver. An antibody that selectively recognizes exon-4 skipping GLT-1 revealed strong signals in Western blots and labelled grey matter astrocytes. We conclude that exon-4 skipping GLT 1 is abundantly expressed in the brain and may represent either a functional glutamate transporter or a modulator of glutamate transporter function.


Cell cycle and immune-related processes are significantly altered in chronic GVHD.
Currently, the pathogenesis of chronic GVHD is unclear. To elucidate the molecular characteristics underlying chronic GVHD, we analyzed the gene expression profiles of 21 mononuclear cell samples from allogeneic hematopoietic stem cell transplantation (HSCT) recipients. Self organizing map (SOM) clustering showed that the entire expression profiles of chronic GVHD samples were clearly different from those of the non-GVHD samples, and significance analysis of microarray (SAM) demonstrated that 120 genes, including PTDSS1, VAV1 and CD3D, were up-regulated, and 5 genes, including calnexin, were down-regulated in GVHD patients. Gene ontology annotation revealed that these genes are related to the phosphorous metabolism and lipid biosynthesis. Quantitative real time polymerase chain reaction (qRT-PCR) experiments validated the up-regulation of PTDSS1, VAV1 and CD3D in separate samples. Pathway-wise global test revealed that differential gene expression in cell cycle and T cell immune-associated pathways were significant between GVHD patients and non-GVHD patients. Seventeen classifier genes selected using a PAM (prediction analysis of microarray) algorithm showed favorable performance (prediction accuracy=0.85) for identifying patients with chronic GVHD. In conclusion, we identified differentially expressed genes and pathways in chronic GVHD patients using microarray analysis, and we also selected diagnostic genes predicting chronic GVHD status.


Novel mutations underlying nephrogenic diabetes insipidus in Arab families.
Nephrogenic Diabetes Insipidus (NDI) is genetically heterogeneous and may be inherited in an X-linked or autosomal recessive manner. We aimed to investigate the molecular basis of NDI among Arab families. Direct sequencing of coding regions for AQP2 and AVPR2 was used to identify underlying mutations. One large deletion required Southern blot analysis and a PCR-based strategy to identify deletion junctions. We identified two novel missense mutations (AQP2:p.Gly100Arg and p.Gly180Ser) in AQP2 and one novel missense mutation (AVPR2:p.Gly122Asp), one previously reported missense mutation (AVPR2:p.Arg137His) and one novel contiguous deletion (AVPR2:c.25 + 273_ARHGAP4o:2650-420del) affecting AVPR2. We also describe evidence of lyonization associated with the novel deletion. Two novel mutations were identified in each of AVPR2 and AQP2 underlying CNDI in Arab families. Identification of these mutations will facilitate early diagnosis of CNDI, counseling of families and provide opportunities for early intervention aimed at reducing morbidity. The presence of affected females and consanguinity, as is often observed in Arab communities should not be used to rule out AVPR2 as a candidate when considering diagnostic testing. Careful observation of phenotypic heterogeneity should be used in referring such families for both AQP2 and AVPR2 molecular genetic testing.


Hepatitis B Virus Infection Is Independently Associated With Advanced Colorectal Adenoma.
Studies on the association of chronic hepatitis B virus (HBV) infection with colonic neoplasm are rare. We aimed to investigate the association between chronic HBV infection and the development of colonic adenoma. One hundred thirty-three patients with chronic HBV infection who underwent colonoscopic examination, were enrolled. A healthy HBV-uninfected group was matched with the HBV group. Those with a previous history of colorectal cancer, inflammatory bowel diseases or colorectal surgery were excluded from both HBV and HBV-uninfected groups. Clinical information and data on age, sex, body mass index, smoking, alcohol consumption and comorbidities were obtained. Advanced adenoma was defined as tubular adenoma ≥10mm, or adenoma with a villous component or high-grade dysplasia. The HBV group had a higher rate of colorectal adenoma and advanced adenoma than the HBV-uninfected group. Patients in the HBV group had larger colorectal polyps than those in the HBV-uninfected group. In the chi square test, HBV DNA positivity was significantly associated with colorectal adenoma (P < 0.001) and advanced adenoma (P = 0.007). HBV infection (odds ratio [OR] = 23.961, 95% CI: 9.400-61.076), diabetes mellitus (OR = 2.633, 95% CI: 1.071-6.473) and age (OR = 1.057, 95% CI: 1.020-1.095) were significantly associated with advanced adenoma in the multivariable logistic regression analysis. Multivariable logistic regression analysis within the HBV group revealed that HBV DNA (OR = 1.236, 95% CI: 1.029-1.485) was associated with advanced adenoma. HBV DNA in patients with HBV infection and HBV infection are independently associated with advanced colorectal adenoma development.


Three-dimensional virtual simulation of alar width changes following bimaxillary osteotomies.
The aim of this study was to evaluate the accuracy of three-dimensional (3D) soft tissue simulation of nose width changes following bimaxillary osteotomies and to identify patient- and surgery-related factors that may affect the accuracy of simulation. Sixty patients (mean age 26 years) who underwent bimaxillary osteotomies participated in this study. Cone beam computed tomography scans were acquired preoperatively and at 1-year postoperative. The 3D hard and soft tissue rendered preoperative and postoperative virtual head models were superimposed, after which the maxilla and mandible were segmented and aligned to the postoperative position. The postoperative changes in alar width were simulated using a mass tensor model (MTM)-based algorithm and compared with the postoperative outcome. 3D cephalometric analyses were used to quantify the simulation error. The postoperative alar width was increased by 1.6±1.1mm and the mean error between the 3D simulation and the actual postoperative alar width was 1.0±0.9mm. The predictability was not correlated to factors such as age, sex, alar cinch suture, VY closure, maxillary advancement, or a history of surgically assisted rapid maxillary expansion. The MTM-based simulation model of postoperative alar width change was found to be reasonably accurate, although there is room for further improvement.


Kinetic studies of elemental mercury adsorption in activated carbon fixed bed reactor.
Activated carbons are suitable materials for Hg(0) adsorption in fixed bed operation or in injection process. The fixed bed tests provide good indication of activated carbons effectiveness and service lives, which depend on the rates of Hg(0) adsorption. In order to correlate fixed bed properties and operation conditions, with their adsorptive capacity and saturation time, Hg(0) adsorption tests were realized in a bench-scale unit, consisted of F400 activated carbon fixed bed reactor. Hg(0) adsorption tests were conducted at 50 degrees C, under 0.1 and 0.35 ng/cm(3) Hg(0) initial concentrations and with carbon particle sizes ranging between 75-106 and 150-250 microm. Based on the experimental breakthrough data, kinetic studies were performed to investigate the mechanism of adsorption and the rate controlling steps. Kinetic models evaluated include the Fick's intraparticle diffusion equation, the pseudo-first order model, the pseudo-second order model and Elovich kinetic equation. The obtained experimental results revealed that the increase in particle size resulted in significant decrease of breakthrough time and mercury adsorptive capacity, due to the enhanced internal diffusion limitations and smaller external mass transfer coefficients. Additionally, higher initial mercury concentrations resulted in increased breakthrough time and mercury uptake. From the kinetic studies results it was observed that all the examined models describes efficiently Hg(0) breakthrough curves, from breakpoint up to equilibrium time. The most accurate prediction of the experimental data was achieved by second order model, indicating that the chemisorption rate seems to be the controlling step in the procedure. However, the successful attempt to describe mercury uptake with Fick's diffusion model and the first order kinetic model, reveals that the adsorption mechanism studied was complex and followed both surface adsorption and particle diffusion.


Chest CT for rapid triage of patients in multiple emergency departments during COVID-19 epidemic: experience report from a large French university hospital.
To assess the diagnostic performances of chest CT for triage of patients in multiple emergency departments during COVID-19 epidemic, in comparison with reverse transcription polymerase chain reaction (RT-PCR) test. From March 3 to April 4, 2020, 694 consecutive patients from three emergency departments of a large university hospital, for which a hospitalization was planned whatever the reasons, i.e., COVID- or non-COVID-related, underwent a chest CT and one or several RT-PCR tests. Chest CTs were rated as "Surely COVID+," "Possible COVID+," or "COVID-" by experienced radiologists. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated using the final RT-PCR test as standard of reference. The delays for CT reports and RT-PCR results were recorded and compared. Among the 694 patients, 287 were positive on the final RT-PCR exam. Concerning the 694 chest CT, 308 were rated as "Surely COVID+", 34 as "Possible COVID+," and 352 as "COVID-." When considering only the "Surely COVID+" CT as positive, accuracy, sensitivity, specificity, PPV, and NPV reached 88.9%, 90.2%, 88%, 84.1%, and 92.7%, respectively, with respect to final RT-PCR test. The mean delay for CT reports was three times shorter than for RT-PCR results (187 ± 148 min versus 573 ± 327 min, p < 0.0001). During COVID-19 epidemic phase, chest CT is a rapid and most probably an adequately reliable tool to refer patients requiring hospitalization to the COVID+ or COVID- hospital units, when response times for virological tests are too long. • In a large university hospital in Lyon, France, the accuracy, sensitivity, specificity, PPV, and NPV of chest CT for COVID-19 reached 88.9%, 90.2%, 88%, 84.1%, and 92.7%, respectively, using RT-PCR as standard of reference. • The mean delay for CT reports was three times shorter than for RT-PCR results (187 ± 148 min versus 573 ± 327 min, p < 0.0001). • Due to high accuracy of chest CT for COVID-19 and shorter time for CT reports than RT-PCR results, chest CT can be used to orient patients suspected to be positive towards the COVID+ unit to decrease congestion in the emergency departments.


Macrophage differentiation inducing factor from human monocytic cells is equivalent to murine leukemia inhibitory factor.
Human macrophage differentiation inducing factor (DIF) can induce differentiation of human myeloid leukemic cells into macrophage-like cells in vitro. A procedure is described for purification of DIF from serum-free human monocytic leukemia THP-1 cell-conditioned medium. The procedure included concentration of a conditioned medium by ultrafiltration, lentil lectin-Sepharose affinity chromatography, and fast protein liquid chromatography using Mono S and Mono Q. DIF-A of pI 9.0 and DIF-B of pI 8.8 were obtained after a final purification with a Mono Q column, and both DIF gave a single peak with a molecular weight of approximately 51,000 determined by gel chromatography. NH2-terminal amino acid analysis of DIF-A showed a noticeable homology with murine leukemia inhibitory factor. Human DIF-A was found to induce maturation of human and murine leukemic cells into both macrophage-like cells with nitro blue tetrazolium reducing activity and phagocytic cells, but was found to suppress proliferation of these leukemic cells.


Beyond the patient: the broader impact of genetic discrimination among individuals at risk of Huntington disease.
We aimed to address gaps in current understanding of the scope and impact of discrimination, by examining a cohort of individuals at-risk for Huntington disease (HD), to describe the prevalence of concern for oneself and one's family in multiple domains; strategies used to mitigate discrimination; and the extent to which concerns relate to experiences. We conducted a cross-sectional survey of 293 individuals at-risk for HD (80% response rate); 167 respondents were genetically tested and 66 were not. Fear of discrimination was widespread (86%), particularly in the insurance, family and social settings. Approximately half of concerned individuals experienced discrimination (40-62%, depending on genetic status). Concern was associated with "keeping quiet" about one's risk of HD or "taking action to avoid" discrimination. Importantly, concern was highly distressing for some respondents (21% for oneself; 32% for relatives). Overall, concerned respondents with high education levels, who discovered their family history at a younger age, and those who were mutation-positive were more likely to report experiences of discrimination than others who were concerned. Concerns were rarely attributed to genetic test results alone. Concern about genetic discrimination is frequent among individuals at-risk of HD and spans many settings. It influences behavioral patterns and can result in high levels of self-rated distress, highlighting the need for practice and policy interventions. © 2012 Wiley Periodicals, Inc.


Twin study of neonatal transient-evoked otoacoustic emissions.
Our knowledge of which physiological mechanisms shape transient evoked otoacoustic emissions (TEOAEs) is incomplete, although thousands of TEOAEs are recorded each day as part of universal newborn hearing-screening (UNHS). TEOAE heritability may explain some of the large TEOAE variability observed in neonates, and give insights into the TEOAE generators and modulators, and why TEOAEs are generally larger in females and right ears. The aim was to estimate TEOAE heritability and describe ear and sex effects in a consecutive subset of all twins that passed UNHS at the same occasion at two hospitals during a six-year period (more than 30 000 neonates screened in total). TEOAEs were studied and TEOAE level correlations compared in twin sets of same-sex (SS, 302 individual twins, 151 twin pairs) and opposite-sex (OS, 152 individual twins, 76 twin pairs). A mathematical model was used to estimate and compare monozygotic (MZ) and dizygotic (DZ) intra-twin pair TEOAE level correlations, based on the data from the SS and OS twin sets. For both SS and OS twin pairs TEOAE levels were significantly higher in right ears and females, compared to left ears and males, as previously demonstrated in young adult twins and large groups of neonates. Neonatal females in OS twin pairs did not demonstrate masculinized TEOAEs, as has been demonstrated for OAEs in young adult females in OS twin pairs. The within-twin pair TEOAE level correlations were higher for SS twin pairs than for OS twin pairs, whereas the within-pair correlation coefficients could not be distinguished from zero when twins were randomly paired. These results reflect heredity as a key factor in TEOAE level variability. Additionally, the estimated MZ within-twin pair TEOAE level correlations were higher than those for DZ twin pairs. The heritability estimates reached up to 100% TEOAE heritability, which is numerically larger than previous estimates of about 75% in young adult twins.


Age effects on the P300 potential and the corresponding fMRI BOLD-signal.
Age has been reported to influence amplitude and latency of the P300 potential. Nevertheless, it is not yet fully understood which brain regions are responsible for these effects. The aim of this study was to investigate age-effects on the P300 potential and the simultaneously acquired BOLD signal of functional MRI. 32 healthy male subjects were investigated using an auditory oddball paradigm. The functional MRI data were acquired in temporal synchrony to the task. The evoked potential data were recorded during the intervals in between MR image acquisitions in order to reduce the influence of the scanner noise on the presentation of the tones and to reduce gradient artifacts. The age-effects were calculated by means of regression analyses. In addition, brain regions modulated by the task-induced amplitude variation of the P300 were identified (single trial analysis). The results indicated an age effect on the P300 amplitude. Younger subjects demonstrated increased parietal P300 amplitudes and increased BOLD responses in a network of brain regions including the anterior and posterior cingulate cortex, the insula, the temporo-parietal junction, the superior temporal gyrus, the caudate body, the amygdala and the parahippocampal gyrus. Single trial coupling of EEG and fMRI indicated that P300 amplitudes were predominantly associated with neural responses in the anterior cingulate cortex, the putamen and temporal brain areas. Taken together, the results indicate diminished neural responses in older compared to younger subjects especially in frontal, temporo-parietal and subcortical brain regions.


Preclinical evaluation of the PI3K-mTOR dual inhibitor PF-04691502 as a novel therapeutic drug in nasopharyngeal carcinoma.
Nasopharyngeal carcinoma (NPC) is common in Southeast Asia and over 40% of NPC tissues have PIK3CA amplification. This study characterized the preclinical activity of a novel potent dual PI3K/mTOR inhibitor, PF-04691502, in five NPC cell lines: CNE-1, HK1, CNE-2, HONE-1 and C666-1, in which all of the cell lines possessed basal and activated expression of Akt and p70S6K. Over 80% inhibition of cell growth in all of these cell lines were achieved after 72 h of PF-04691502 incubation and their IC50 were in hundred nanomolar range. CNE-2, HK1 and HONE-1 were selected to further evaluate the effect of PF-04691502 on cell cycle, apoptosis and Akt downstream signaling. PF-04691502 induced G0/G1 cell cycle arrest and apoptosis at 24 h incubation and it significantly abrogated Akt and its downstream signaling by suppressing the expression of p-mTOR, p-p70S6K, p-Akt(S473, T308), p-S6 and p-4E-BP1, suggesting its effectiveness in inhibition of translation and protein synthesis. Anti-proliferation was also observed in 3D culture system and spheroids formation of NPC cell line HONE-1-EBV was strongly inhibited by PF-04691502. Antitumor activity was observed in CNE-2 xenograft in 2 weeks of 10 mg/kg PF-09641502 treatment to tumor bearing athymic nude mice. Both tumor volume and weight in treatment group were significantly lower than those in vehicle group while no obvious body weight decrease was found, suggesting this working dose was effective and well-tolerated. Additive effects were observed in combination of PF-09641502 with either cisplatin or paclitaxel. There were no synergistic effect observed in drug combination but PF-09641502 alone was effective in treating cisplatin resistant cell lines as compared to its parental control. The beneficial effects of PF-09641502 in both in vitro and in vivo studies for NPC warrant a further investigation.


Design, synthesis, biological and structural evaluation of functionalized resveratrol analogues as inhibitors of quinone reductase 2.
Resveratrol (3,5,4'-trihydroxylstilbene) has been proposed to elicit a variety of positive health effects including protection against cancer and cardiovascular disease. The highest affinity target of resveratrol identified so far is the oxidoreductase enzyme quinone reductase 2 (QR2), which is believed to function in metabolic reduction and detoxification processes; however, evidence exists linking QR2 to the metabolic activation of quinones, which can lead to cell toxicity. Therefore, inhibition of QR2 by resveratrol may protect cells against reactive intermediates and eventually cancer. With the aim of identifying novel inhibitors of QR2, we designed, synthesized, and tested two generations of resveratrol analogue libraries for inhibition of QR2. In addition, X-ray crystal structures of six of the resveratrol analogues in the active site of QR2 were determined. Several novel inhibitors of QR2 were successfully identified as well as a compound that inhibits QR2 with a novel binding orientation.


Day-One Pain Reductions After Hip and Knee Replacement When Buprenorphine-Clonidine-Dexamethasone Is Added to Bupivacaine Nerve/Plexus Blocks: A Randomized Clinical Trial.
To compare pain outcome reports of patients undergoing hip or knee replacement who received single-injection nerve/plexus blocks with plain bupivacaine (BPV) with those of patients who received injections of buprenorphine-clonidine-dexamethasone (BCD) admixed with BPV. Prospective, parallel-arm, randomized, double-blind trial. A single veterans' hospital. Ninety-eight veterans scheduled for total hip or knee replacement surgery with spinal as the primary anesthetic. Participants were randomized to BPV-BCD or plain BPV groups. They underwent nerve/plexus blocks in the L2-L4 and L4-S3 distributions in advance of joint replacement surgery. The primary outcome was change in pain from baseline during the postoperative day, as assessed by the total pain score on the short-form McGill Pain Questionnaire-v2 (SF-MPQ-2). Secondary outcomes were pain during movement, pain interference, range of motion, mobility, and quality of recovery. On postoperative day one, the SF-MPQ-2 total score for the BPV-BCD group demonstrated greater pain reduction than that of the plain BPV group (mean difference 1.8 points, 95% confidence interval 0.6 to 3.0, P = 0.003). The BPV-BCD group also had larger reductions in pain during movement in the surgical joint and less pain interference, along with increased range of hip and knee flexion, compared with the plain BPV group. Outcomes of mobility and quality of recovery were not different between groups. Preoperative BPV-BCD blocks in the L2-L4 and L4-S3 nerve distributions for hip and knee replacements led to less pain on postoperative day one and increased knee and hip range of motion, compared with plain BPV blocks. ClinicalTrials.gov ID NCT02891798.


Retrograde lymphatic spread and ischemia of the forearm and hand--rare complications of carcinoma of the breast.
A case of ischemia of the forearm and hand with resulting necrosis three years following simple mastectomy and radiation therapy for breast carcinoma is reported. Histologic sections of the mid-forearm amputation specimen showed extensive lymphatic involvement by tumor. The latter process is judged to have been one of retrograde lymphatic extension.


Exercise after You Eat: Hitting the Postprandial Glucose Target.
We discuss a novel hypothesis: the effect size of postmeal exercise for attenuating postprandial glucose will be a function of the exercise bout vs. the size of the postprandial glucose response, specifically peak and duration of the postprandial glucose excursion.


Vertical root fractures in crowned teeth: a report of 32 cases.
To clarify the mode of root fracture in 32 endodontically treated teeth that resulted in vertical root fracture (VRF) despite the provision of a crown for fracture prevention. The teeth included in the study underwent root canal treatment and were restored with crowns without the use of an intracanal post. All teeth underwent photo documentation. Clinical signs and symptoms as well as the patients' ages were recorded. The diameters of the roots and the root canal fillings were measured on radiographs. Fractured roots were investigated using a scanning electron microscope (SEM). The mean patient age was 60.2 years. The VRFs in the group occurred most frequently in the mesial roots of the mandibular first molars. The fracture was predominantly buccolingual. The mean ratio measured between the diameters of the root canal obturations and the outer root surfaces in the radiographs was 33.2%. SEM evaluation showed that most fractures ran from the inner to the outer root surface, and the cracks predominantly ran apical to coronal. Endodontically treated and crowned teeth without root canal posts can develop VRFs. The cracks start at the root canal wall in the apical part of the root and extend toward the outer root surface and coronally. Further investigation is necessary to determine the possible causes and evidence of fracture development.


The complete characterization of a rhodium Lewis acid-dipolarophile complex as an intermediate for the enantioselective catalytic 1,3-dipolar cycloaddition of C,N-diphenylnitrone to methacrolein.
The 1,3-dipolar cycloaddition reaction of C,N-diphenylnitrone with methacrolein is efficiently catalyzed by the rhodium diphosphine compound (SRh,RC)-[(eta5-C5Me5)Rh(R-Prophos)(H2O)](SbF6)2 [R-Prophos = (R)-(+)-1,2-bis(diphenylphosphino)-propane, 1.SbF6]; the asymmetric catalytic process occurs with reversal of regioselectivity, perfect endo selectivity, and up to 92% ee. The complete (NMR and X-ray analysis) characterization of the involved intermediate (SRh,RC)-[(eta5-C5Me5)Rh(R-Prophos)(methacrolein)](SbF6)2 (7.SbF6) allows us to interpret the observed selectivity.


Bronchoalveolar lavage examined by solid phase microextraction, gas chromatography--mass spectrometry and selected ion flow tube mass spectrometry.
Samples (210 in total) of broncholaveolar lavages (BALs), obtained from patients hospitalized with pneumonia in various departments of two hospitals, were analysed using the method of solid phase microextraction-gas chromatography (SPME-GC) with FID detection. Up to 20% (9% unequivocally, 11% probably) of these samples was found to contain volatile fatty acids (VFAs) in the series from acetic acid to heptanoic acid. Importantly, the presence of these acids indicates the presence of fermenting anaerobic bacteria, which were not detected by the conventional microbiological examination. Other compounds, namely the heptanol and cyclohexanone, were also detected by this method in some samples. Cyclohexanone occurred almost exclusively in samples from patients receiving intensive care with mechanical ventilation, and is suspected to originate from plastic parts of ventilators. Selected representative samples were also analysed using further methods, namely gas chromatography-mass spectrometry (GC-MS) of native and silylated samples, and selected ion flow tube mass spectrometry (SIFT-MS). These methods confirmed the identities of above mentioned compounds, and detected numerous other compounds tentatively identified as various alcohols, aldehydes, ketones, esters and hydrogen cyanide, HCN. Most of these compounds occurred in small amounts and their origin and diagnostic significance remains uncertain, except, that is, for the HCN, which indicates the presence of Pseudomonas aeruginosa.


Pure melanocyte cultures: differential serum requirements of guinea pig keratinocytes and melanocytes in primary epidermal cell cultures.
Contrary to melanocytes guinea pig keratinocytes do not attach and grow in primary epidermal cell cultures if plated in media containing guinea pig serum. This phenomenon is based on the low content or lack of guinea pig serum of a factor(s) promoting keratinocyte attachment. This factor, which is contained in fetal calf serum, binds to the keratinocyte cell surface and can be removed by trypsin. Guinea pig epidermal cell cultures plated in medium containing guinea pig serum therefore lead to pure or almost pure melanocyte cultures.


Assessment of intraoperative bleeding in reverse shoulder arthroplasty - with or without a stem.
The functional results of stemless reverse shoulder prostheses are similar to those with stems. However, the operative time and the bleeding appear less significant because of the absence of humeral reaming. To date, the data amongst the literature regarding this subject is limited. Thus, we report a retrospective evaluation on these 2 types of prostheses by assessing their respective intraoperative blood loss. Reverse shoulder arthroplasty without a stem leads to less blood loss, compared to arthroplasty with a stem. Twenty-three patients underwent an operation for a stemless prosthesis, while 37 patients had a prosthesis with a stem. The hemoglobin was measured preoperatively, as well as postoperatively. Drainage of the operative site was maintained for two to three days. In the stem group, the preoperative hemoglobin was 14g/dL (11.7-16.6), while it was 13.1g/dL (11-15.8) in the stemless group. The intraoperative bleeding reached 223cm3 (80-530), with an operative duration of 81minutes (40-110) in the stemless group, compared to 260cm3 (50-1000) and 92minutes (33-110) in the stem group. On the first day postoperatively, 333cm3 (20-570) of blood had been collected by drainage for the stemless group, compared to 279cm3 (40-550) in the stem group. The amount decreased the second day, with 139cm3 (20-510) and 129cm3 (0-750) respectively. There was no difference between the two groups regarding the postoperative hemoglobin level (11g/dL). There is no significant difference concerning the blood loss between reverse shoulder replacements with and without stems. III Retrospective case control study.


Inheritance and perinatal consequences of inherited thrombophilia in Greece.
To investigate the impact of inherited thrombophilic factors on the gestational outcome of unselected pregnant women. A total of 392 women with spontaneous pregnancy were investigated for Factor V Leiden, prothrombin G20210A, and MTHFR C677T mutations. Adverse pregnancy outcomes were recorded. Thrombophilic genotypes were significantly higher in women with placental abruption. Heterozygocity for Factor V Leiden increased the risk for placental abruption 9.1 times. The MTHFR T677T genotype increased the risk for placental abruption 4.8 times despite folate supplements, and normal serum folate and B(12) levels. Women with inherited thrombophilia and previous obstetric complications were at significant risk for complications in a subsequent pregnancy (P<0.05). Women with placental abruption should be screened for thrombophilic factors and plasma homocysteine should be measured. Subgroups of women with inherited thrombophilia and obstetric complications might benefit from prophylactic anticoagulation in subsequent pregnancies.


Carbon nanotube-enhanced DNA biosensor for DNA hybridization detection using manganese(II)-Schiff base complex as hybridization indicator.
A Mn(II) complex, MnL (L=sodium (E)-3-((1-carboxyethylimino)methyl)-4-hydroxybenzenesulfonate), was synthesized and characterized using elemental analysis and IR spectroscopy. Cyclic voltammetry (CV) and fluorescence spectroscopy were used to investigate the interaction between MnL and salmon sperm DNA. It was revealed that MnL presented high electrochemical activity on glassy carbon electrode (GCE), and it could be intercalated into the double helices of double-stranded DNA (dsDNA). Using MnL as the hybridization indicator, a novel and sensitive electrochemical DNA biosensor based on multiwall carbon nanotubes functionalized with carboxyl groups (MWCNTs-COOH, on which DNA probes were covalently immobilized) was prepared. The target single-stranded DNA (ssDNA) could be quantified ranging from 6.7 x 10(-10)M to 8.4 x 10(-9)M with good linearity (r=0.9922). A detection limit of 1.4 x 10(-10)M (3 sigma, n=9) was achieved.


Pulmonary edema and hydrothorax in swine produced by fumonisin B1, a toxic metabolite of Fusarium moniliforme.
Pulmonary edema and hydrothorax were observed in mature swine that died approximately 5 days after consuming corn screenings. These postmortem observations were reproduced in younger swine (16-24 kg) that died within 1 week when fed the corn screenings under experimental conditions. Additionally, pulmonary edema and hydrothorax occurred in a pig (7.1 kg) that died after receiving 4 daily intravenous injections of fumonisin B1. A fungus was isolated from the corn screenings that is identical to Fusarium moniliforme MRC-826 in colony morphology and under microscopic examination.


Hemodynamic performance and clinical consequences of aortic valve replacement with 21-mm sized pericardial bioprostheses.
Aortic valve replacement (AVR) with a 21-mm sized bioprothesis is still discussed controversially. Since better results have been reported for pericardial valves, the aim of the current study was to analyze the hemodynamic performance as well as clinical parameters in our patients and to compare pericardial and standard porcine valves in particular. 342 patients underwent AVR with a bioprosthesis between 1987 and 2000. A 21 mm prosthesis was used in 39 patients (group S), while 303 patients received at least a 23-mm sized valve (group L). Group S was further divided into 19 patients with a pericardial valve (group S1) and 20 patients with a standard porcine valve (group S2). The hemodynamic and clinical parameters were studied in all three groups. The peak and mean transprosthetic gradients were significantly lower in the pericardial group than in the porcine group, particularly between patients with 21 mm valves (peak/mean: S1: 24 +/- 9/20.8 +/- 6.5 mm Hg vs. S2: 38 +/- 15/33 +/- 9 mm Hg, p < 0.05) at discharge. We could also observe that the peak transprosthetic gradient 7 days postoperatively was not significantly higher in patients with a 21 mm pericardial valve compared to group L patients. Comparing clinical parameters, we found significantly more cerebral ischemic events, a prolonged mechanical ventilation, a higher mortality and a longer stay in hospital in the group S2 compared to the group S1. The current study shows that pericardial valves perform well, particularly in patients with small aortic roots. Postoperative hemodynamics and clinical results were better than for comparable standard porcine valves. As the outcome of patients with a 21 mm pericardial valve was no worse than that in patients with bigger valves, enlarging procedures for the aortic root are not necessary in the majority of these patients.


[Effects of 10-hydroxycamptothecin on induced chromosome aberrations in Chinese hamster ovary cells and micronuclei in mouse bone marrow and fetal liver].
10-Hydroxycamptothecin (HC) is a new antitumor principle isolated from Camptotheca acuminata indigenous to China. The genetic toxicity of HC was assessed by mouse bone marrow and transplacental micronucleus test as well as Chinese hamster ovary cell chromosomal aberrations. All of these tests showed positive results. The highest rate of chromosomal aberrations was 83% at 0.125 microgram/ml for 48 h. The number of micronucleated polychromatic erythrocytes in bone marrow of mice was remarkably increased in 19.8% cells at 12.5 mg/kg for 24 h. The micronucleus formation was most often seen at 16 h after im HC in fetal liver and 24 h in maternal bone marrow. The peaks were 36 +/- 19 and 31 +/- 10%, respectively. The results from in vivo and in vitro suggest HC is a mutagen, furthermore, a transplacental mutagen in mouse.


Patterns of resource use within a population of xanthid crabs occupying bryozoan colonies.
Heads, i.e. colonies, of the bryozoan Schizoporella pungens are discrete resource units which protect the xanthid crab Pilumnus sayi from predation. The heads were spatially clumped, temporally stable, and differed both qualitatively and quantitatively. The number of adult P. sayi was equal to the number of heads with preferred characteristics, yet a relative shortage of this resource is suggested with concomitant intraspecific competition. Adult crabs occupied heads individually except for infrequent cases of double occupancy by a male and female. Further spatial pattern among crabs was consistent with a hypothesis that males defend clumps of heads occupied by females.


Nitric oxide-dependent hypotensive effects of wax gourd juice.
The wax gourd (Benincasa hispida (Thunb) Cong.) is a long-season vegetable that has been used in traditional Chinese medicine to treat high blood pressure. However, precise details of its effect and the mechanism of action involved are still lacking. Ten-fold-condensed wax gourd juice was used for the experiments. We measured (1) blood pressure of anesthetized normal Wistar rats in vivo, (2) isolated rat aortic contraction and relaxation, and (3) nitric oxide production from cultured porcine endothelial cells. The rats mentioned had not been treated with the investigational medicine. Intravenous injection of the juice produced a dose-dependent decrease in blood pressure. Treatment with the juice induced concentration-dependent relaxation of isolated rat aortic rings that had been precontracted with noradrenaline. The relaxation induced by the juice was strongly inhibited by treatment with the nitric oxide (NO) synthase inhibitor N(G)-nitro-l-arginine methyl ester hydrochloride (l-NAME) or endothelial denudation. Treatment with the juice produced NO from cultured porcine aortic endothelial cells. This NO production was significantly inhibited by l-NAME. The present findings suggest that wax gourd juice exerts a hypotensive effect via endothelium-dependent vasodilation. The main endothelium-derived relaxing factor involved might be NO.


Trends of nitrogen in air and precipitation: model results and observations at EMEP sites in Europe, 1980--2003.
We analyze trends of some nitrogen compounds using long-term measurements and results from the EMEP (co-operative programme for monitoring and evaluation of the long-range transmissions of air pollutants in Europe) chemical transport model at EMEP sites. We find statistically significant declines at the majority of sites for NH(x) (sum of ammonia and ammonium) in air and for nitrate and ammonium in precipitation, but only at a few sites for xNO3 (sum of nitrate and nitric acid) in air. Model calculations and measurements give similar results. We demonstrate that the lack of trends for xNO3 in air at least partly can be attributed to a shift in the equilibrium between nitric acid and ammonium nitrate towards particulate phase, caused by reductions in the sulfur dioxide emissions.


Hepatitis B virus genotype D prevails in patients with persistently elevated or normal ALT levels in Turkey.
The clinical relevance of hepatitis B virus (HBV) genotypes are poorly understood and it is unclear if the prevalence of HBV genotypes differs with the various clinical features of HBV carriers. The aim of our study was to examine the prevalence of the HBV genotype in a group of patients with chronic hepatitis B, compared to a group with chronic inactive hepatitos B surface antigen (HbsAg) carriers. HBV genotypes were determined in 32 patients with chronic hepatitis B and in 12 chronic inactive HBsAg carriers. 35 males and nine females with a mean age of 33.95 +/- 13.04 were studied. Serum samples were examined for the presence of HBV DNA by polymerase chain reaction (PCR). Samples negative in first round PCR were further amplified with nested PCR. The PCR product was sequenced with the Cy5/5.5 dye primer kit on a Long Read Tower automated DNA sequencer. HBV DNA was detectable in 29 (66%) and 44 (100%) patients by the PCR with universal primers and nested-PCR, respectively. All patients were found to be infected with HBV genotype D. Genotype D was the only detected type found in different clinical forms of chronic HBV infection, in all hepatitis B e antigen (HbeAg)-positive and negative patients, in all patients who had elevated or normal alanine transaminase (ALT) levels and in all ages. In the present study we could not find any association between genotype D and distinct clinical phenotypes. Genotype D is the predominant type among hepatitis B carriers residing in our region and is not associated with more severe liver diseases. This genotype did not influence clinical manifestations in carriers with chronic hepatitis B virus infection. However, additional large-scale longitudinal studies are needed to find the relationship of HBV genotypes to liver disease severity and clinical outcomes.


Vibrational disorder and densification-induced homogenization of local elasticity in silicate glasses.
We report the effect of structural compaction on the statistics of elastic disorder in a silicate glass, using heterogeneous elasticity theory with the coherent potential approximation (HET-CPA) and a log-normal distribution of the spatial fluctuations of the shear modulus. The object of our study, a soda lime magnesia silicate glass, is compacted by hot-compression up to 2 GPa (corresponding to a permanent densification of ~ 5%). Using THz vibrational spectroscopic data and bulk mechanical properties as inputs, HET-CPA evaluates the degree of disorder in terms of the length-scale of elastic fluctuations and the non-affine part of the shear modulus. Permanent densification decreases the extent of non-affine elasticity, resulting in a more homogeneous distribution of strain energy, while also decreasing the correlation length of elastic heterogeneity. Complementary 29Si magic angle spinning NMR spectroscopic data provide a short-range rationale for the effect of compression on glass structure in terms of a narrowing of the Si-O-Si bond-angle and the Si-Si distance.


Birth by cesarean section is associated with elevated neonatal plasma levels of dimethylarginines.
This study was undertaken to compare the effects of vaginal delivery and cesarean section on the L-arginine-nitric oxide system by measuring levels of L-arginine, an endogenous nitric oxide synthase antagonist asymmetric dimethylarginine (ADMA), and symmetric dimethylarginine (SDMA) in the cord blood and postnatally. Plasma samples were obtained from the umbilical vein and artery at birth and from peripheral venous blood on the second postnatal day in 30 full-term newborn infants: 10 born vaginally and 20 born by cesarean section. After vaginal delivery, ADMA concentration was higher in the umbilical vein than in the umbilical artery (mean 1.06 vs 0.90 µmol/L [P = 0.027]); and ADMA level fell after birth to 0.66 µmol/L on the second postnatal day (P = 0.007 vs umbilical artery). Newborns born by cesarean section had similar ADMA levels in umbilical arterial and venous blood, 1.19 and 1.18 µmol/L, and the ADMA level fell to 0.84 µmol/L by the second postnatal day (P < 0.001). Vaginal birth induced neither significant umbilical venoarterial difference nor a postnatal fall in SDMA. After cesarean section, SDMA was essentially the same in umbilical vein, umbilical artery and postnatal peripheral vein samples. At 2 days of age, both ADMA and SDMA levels stayed higher in infants born by cesarean section than in vaginally born infants. ADMA level falls after both vaginal and cesarean birth, whereas SDMA level does not. The higher ADMA level after cesarean birth compared with vaginal birth may contribute to decreased nitric oxide production and bioavailability in neonatal vascular beds.


Pheochromocytoma and Paraganglioma: Genetics, Diagnosis, and Treatment.
Pheochromocytomas (PCCs) and paragangliomas (PGLs) are rare but unique neuroendocrine tumors. The hypersecretion of catecholamines from the tumors can be associated with high morbidity and mortality, even when tumors are benign. Up to 40% of PCCs/PGLs are associated with germline mutations in susceptibility genes. About one-quarter are malignant, defined by the presence of distant metastases. Treatment options for unresectable metastatic disease, including chemotherapy, (131)I-MIBG, and radiation, can offer limited tumor and hormone control, although none are curative. This article reviews the inherited genetics, diagnosis, and treatment of PCCs and PGLs.


Left ventricular dysfunction in a child with asymptomatic pre-excitation: Cure with catheter ablation.
Wolff-Parkinson-White syndrome, is known to cause left ventricular dysfunction or dilated cardiomyopathy secondary to sustained tachycardia in infants and children. However, left ventricular dysfunction secondary to pre-excitation related abnormal ventricular activation has been reported in a limited number of cases. This condition should be recognized early, as catheter ablation of the accessory pathway can permit rapid ventricular function improvement. In this paper, we present a 2.5-year-old patient diagnosed with tachycardia-free Wolff-Parkinson-White syndrome with a right free wall accessory pathway and depressed cardiac function, whose left ventricular function is completely restored after successful catheter cryoablation.


Two assessments of American usage frequencies for ninety-seven sentence complement-taking nouns.
Two studies documented the American English usage frequencies for ninety-seven nouns that occur with sentence complements (e.g., "rumor," "The rumor that the student cheated..."). An established usage preference methodology was used (see Connine, Ferreira, Jones, Clifton, & Frazier, 1984; Kennison, 1999). In Study 1, participants completed short fragments that did not contain the overt complementizer "that," as in "The rumor..." In Study 2, participants completed short sentence fragments that contained the overt complementizer, as in "The rumor that..." The results showed that when the complementizer was absent (Study 1), bare (i.e., unmodified) usages and prepositional phrase usages were frequently observed. In contrast, sentence complement and relative clause usages were rarely observed. When the complementizer was present (Study 2), the frequency of sentence complement and relative clause usages varied. The estimates of usage frequency obtained in these studies are intended to be a resource for language comprehension researchers.


Medication adherence to rivaroxaban and dabigatran in patients with non-valvular atrial fibrillation: a meta-analysis.
Several real-world studies have evaluated adherence to direct acting oral anticoagulants in patients with non-valvular atrial fibrillation (NVAF); however, these studies have not been systematically summarized. We performed a meta-analysis to compare adherence to rivaroxaban versus dabigatran therapy in United States (US) patients with NVAF in a real-world setting. Medline and Scopus were searched from January 2010 to August 2018 using keywords and MeSH terms related to adherence and oral anticoagulants. We included real-world studies of US adults with NVAF comparing adherence to dabigatran and rivaroxaban. Studies evaluating adherence by a measure other than proportion of days covered (PDC) were excluded. The proportion of patients with a PDC ≥ 80 (a commonly utilized definition of adherence) served as the primary outcome of interest. We conducted meta-analysis of non-overlapping studies using the Hartung-Knapp random-effects model to estimate risk ratios (RRs) with corresponding 95% confidence intervals (CIs). We included 5 studies evaluating 80,230 patients (range 2667-22,571). Median follow-up across studies was 6 months (range 3-12 months). The proportion of patients with a PDC ≥ 80 ranged from 59.5 to 83.5% for rivaroxaban users and 57.3 to 78.3% for dabigatran users. Upon meta-analysis, rivaroxaban use was associated with increased adherence compared with dabigatran use (RR 1.08; 95% CI 1.03-1.12; I2 = 88%). In conclusion, rivaroxaban was associated with increased adherence when compared to dabigatran in ~ 80,000 patients in a real-world setting. Possible explanations for this include dosing frequency or patient tolerance.


Usefulness of a linear analog scale questionnaire to measure health-related quality of life in elderly patients with chronic obstructive pulmonary disease.
To examine the validity, discriminatory ability, and responsiveness of health-related quality-of-life (HRQoL) questionnaires using a linear analog scale (Quality of Life (QOL) scale) for chronic obstructive pulmonary disease (COPD). Cross-sectional and longitudinal. Outpatient. One hundred two elderly subjects with mild to severe COPD. Scores on the QOL scale, the St. George's Respiratory Questionnaire (SGRQ), and the 36-item Short-Form Health Survey questionnaire (SF-36) and various clinical parameters were recorded. The correlations between these QOL questionnaires and various clinical parameters were then examined. The responses of 31 elderly COPD patients to the QOL scale and the SGRQ before and 3 months after the completion of a comprehensive pulmonary rehabilitation program were compared longitudinally. On cross-sectional study, the QOL scale showed a significant correlation with the total score and three components of the SGRQ. The QOL scale correlated significantly with all components of the SF-36, but the total SGRQ score correlated with only six components of the SF-36, excluding vitality and the mental health index. Both the QOL scale and the total score of the SGRQ correlated significantly with the oxygen cost diagram (OCD), Morale scale, 6-minute walking distance (6MWD), forced expiratory volume in 1 second, and instrumental activities of daily living (IADL) score. When subjects were divided into three groups according to disease severity (mild, moderate, severe) using American Thoracic Society guidelines, the total SGRQ score discriminated between the three groups. The QOL scale could not discriminate between mild and moderate or moderate and severe. On longitudinal study, 3 months after finishing the comprehensive pulmonary rehabilitation program, the QOL scale, the SGRQ, 6MWD, and OCD all showed significant improvement. The difference in the QOL scale after the comprehensive pulmonary rehabilitation program showed a significant correlation with changes in the SGRQ total score and the OCD but not with the 6MWD. The QOL scale is similar to more-complex questionnaires such as the SGRQ in terms of validity and responsiveness for evaluating disease-specific HRQoL in elderly COPD patients. In clinical settings, the QOL scale, as a simple questionnaire, may be useful for disease-specific HRQoL assessments in elderly COPD patients.


Commensal bacteria mediated defenses against pathogens.
Commensal bacterial communities residing within the intestinal lumen of mammals have evolved to flourish in this microenvironment. To preserve this niche, commensal bacteria act with the host to prevent colonization by invasive pathogens that induce inflammation and disrupt the intestinal niche commensal bacteria occupy. Thus, it is mutually beneficial to the host and commensal bacteria to inhibit a pathogen's ability to establish an infection. Commensal bacteria express factors that support colonization, maximize nutrient uptake, and produce metabolites that confer a survival advantage over pathogens. Further, commensal bacteria stimulate the host's immune defenses and drive tonic expression of anti-microbial factors. In combination, these mechanisms preserve the niche for commensal bacteria and assist the host in preventing infection.


Treefall gaps versus forest understory as environments for a defoliating moth on a tropical forest shrub.
The moth Zunacetha annulata (Dioptidae) is a specialist on the understory shrub Hybanthus prunifolius (Violaceae) in the forest of Barro Colorado Island (BCI), Panama. The larvae, which are capable of defoliating entire shrubs, concentrate their attack upon the small minority of H. prunifolius individuals that grow in treefall gaps. Field experiments demonstrated that larval growth rates were 37% higher, and weights at pupation 25% higher, on shrubs in gaps than on shrubs in the understory. In a common environment in the laboratory, growth rates of larvae were 23% higher on foliage taken from shrubs in gaps than on foliage from shrubs in the understory.However, larvae grown in a temperature regime simulating that of gaps did not grow faster than larvae in an understory regime, when the two groups were reared in growth chambers on foliage taken from the same shrubs. In the field, predation appeared higher in gaps: experimental groups of larvae survived at rates of 65% per day in gaps and 78% per day in the understory. Quality of foliage, and not direct effects of the environment, appears to be responsible for the observed pattern of defoliation by this moth.


Toxicokinetics of ochratoxin A in several species and its plasma-binding properties.
The toxicokinetic profile of ochratoxin A was studied after the oral or intravenous administration of 50 ng/g b.w. to fish, quail, mouse, rat and monkey. The elimination half-life varied from 0.68 h after oral administration to fish, up to 840 h after intravenous administration to monkey. The distribution volume ranged from 57 ml/kg in fish to 1500 ml/kg in quail. The plasma clearance was most rapid in quail and fish, 72 and 58 ml/kg.h, respectively, while it was only 0.17 ml/kg.h in monkey. The bioavailability was as low as 1.6% in fish but as high as 97% in mouse. The binding abilities of ochratoxin A to plasma proteins were also studied. From these data we calculated the free fraction of toxin in plasma, which we found to be less than 0.2% in all species investigated (including man) except fish. A similar but smaller investigation on the toxicokinetics and binding properties of ochratoxin B was also performed. Ochratoxin B was more readily eliminated and had a lower affinity for plasma proteins, which partly may explain its lower toxicity.


Impression membrane for the diagnosis of microbial keratitis.
To evaluate a corneal impression membrane (CIM) for isolation of bacteria, fungi and acanthamoeba in suspected microbial keratitis. Consecutive patients presenting with suspected microbial keratitis were included. For each patient, samples were collected in a random order using a surgical blade and a 4-mm-diameter polytetrafluoroethylene CIM disc, and transported in brain heart infusion broth. Risk factors, best corrected visual acuity (BCVA), size, location, depth and healing time of the ulcer were recorded. The microbial isolation rate was used to compare sampling methods. 130 patients were included (mean age 62.6 years, SD 19.0). An antimicrobial had been used prior to presentation in 36 (27.7%) patients. Mean major and minor ulcer diameters were 2.1 mm (SD 2.0) and 1.6 mm (SD 1.7). Mean healing time was 12.4 days (SD 13.6). BCVA at presentation and following healing was 0.7 (SD 0.7) and 0.62 (SD 0.7) (p=0.34). There were 66 isolates (50.8%); 53 (40.8%) using a CIM and 35 (26.9%) using a blade (p=0.02). ITALIC! Staphylococcus aureus and coagulase-negative staphylococci were the commonest isolates. Isolation rate was not influenced by organism type, although in four cases ITALIC! Acanthamoeba spp. were isolated; three using CIM and one a blade. In this study, the isolation of microorganisms from cases of suspected microbial keratitis was significantly higher using a CIM than a surgical blade. A CIM may be a useful alternative or addition for sample collection in microbial keratitis.


Persistent gastroesophageal reflux disease after antireflux surgery in children: I. immediate postoperative evaluation using extended esophageal pH monitoring.
There is a paucity of quantitative and reproducible follow-up data on childhood operations for gastroesophageal reflux disease (GERD). With the development of minimally invasive techniques for antireflux operations in children, there is a need to quantitatively determine immediate outcomes for such operations performed by laparotomy for comparison. A retrospective review of 385 children (age range, 1 week to 15 years) who had a primary antireflux operation in a Children's or University Hospital performed by laparotomy between 1983 and 1997, and who also had an extended esophageal pH study performed within the first 12 postoperative weeks, was conducted. The operations performed included Nissen fundoplication (n = 135), Thal fundoplication (n = 195), and Boerema gastropexy (n = 55). An immediate postoperative failure of the operation to control GERD was defined as an abnormal esophageal pH score persisting up to the twelfth postoperative week. Eleven patients (2.9%) were classified as having an immediate postoperative failure of their operation to control GERD. An additional three patients had an abnormal esophageal pH score 2 weeks postoperatively, which subsequently reverted to a normal esophageal pH score by 12 weeks. The immediate postoperative failure rate was 1.5% (2 of 135) for the Nissen fundoplication, 1.5% (3 of 195) for the Thal fundoplication, and 10.9% (6 of 55) for the Boerema gastropexy. A higher failure rate (five patients, 36%) was seen for the first 14 patients who underwent a Boerema gastropexy during the learning curve period for this operation before 1985, and by excluding these patients the failure rate was 2.4% (1 of 41) after 1985. There was no significantly increased probability of immediate postoperative failure in patients with central nervous system disorders, prematurity, repaired esophageal atresia, or gastric emptying abnormalities. Only 5 (36%) of the 14 children with persisting symptoms suggestive of GERD had immediate postoperative failure of their operation. Extended esophageal pH monitoring during the first 12 postoperative weeks is a helpful tool to assess the immediate outcome of antireflux operations in children because clinical symptoms alone may be unreliable. The immediate failure rate for an antireflux operation performed in children by laparotomy is very low and seems to be unaffected by comorbid factors.


Intrahepatic assembly of very low density lipoproteins. Rate of transport out of the endoplasmic reticulum determines rate of secretion.
To identify the rate-limiting step(s) in the hepatic production of very low density lipoproteins (VLDL), we investigated the intracellular distribution and rate of intracellular transport of de novo synthesized apolipoprotein B (apoB). For all secretory proteins examined (i.e. albumin, large molecular weight apoB, and small molecular weight apoB) the rough and smooth microsomes contained the majority of intracellular de novo synthesized protein, while the Golgi subfraction contained 10% or less. Pulse-chase analysis of the intracellular movement of apoB and albumin showed that the first order rate constant (in terms of half-life) describing the rate of movement out of the smooth and rough microsomes determined the overall rate of movement out of the cell. These data suggest that movement out of the endoplasmic reticulum, the site where VLDL is assembled, determines the overall rate of secretion. Furthermore, compared to albumin, the rate of intracellular transport of apoB was approximately two times slower suggesting that processing unique to VLDL apoB occurring in the endoplasmic reticulum was responsible. Additional studies show that essentially all of the de novo synthesized 35S-labeled albumin (produced from a pulse of [35S]methionine) lost from the cell during the chase period could be recovered in the culture medium. In contrast, much less of large molecular weight apoB (36%) and small molecular weight apoB (60%) was recovered in the culture medium. Since these cultured rat hepatocytes do not take up or degrade newly secreted apoB, these data suggest that a significant amount of apoB is degraded intracellularly.


Therapeutic mTOR blockade in systemic autoimmunity: Implications for antiviral immunity and extension of lifespan.
The mechanistic target of rapamycin (mTOR) pathway integrates metabolic cues into cell fate decisions. A particularly fateful event during the adaptive immune response is the engagement of a T cell receptor by its cognate antigen presented by an antigen-presenting cell (APC). Here, the induction of adequate T cell activation and lineage specification is critical to mount protective immunity; at the same time, inadequate activation, which could lead to autoimmunity, must be avoided. mTOR forms highly conserved protein complexes 1 and 2 that shape lineage specification by integrating signals originating from TCR engagement, co-stimulatory or co-inhibitory receptors and cytokines and availability of nutrients. If one considers autoimmunity as the result of aberrant lineage specification in response to such signals, the importance of this pathway becomes evident; this provides the conceptual basis for mTOR inhibition in the treatment of systemic autoimmunity, such as systemic lupus erythematosus (SLE). Clinical trials in SLE patients have provided preliminary evidence that mTOR blockade by sirolimus (rapamycin) can reverse pro-inflammatory lineage skewing, including the expansion of Th17 and double-negative T cells and plasma cells and the contraction of regulatory T cells. Moreover, sirolimus has shown promising efficacy in the treatment of refractory idiopathic multicentric Castleman disease, newly characterized by systemic autoimmunity due to mTOR overactivation. Alternatively, mTOR blockade enhances responsiveness to vaccination and reduces infections by influenza virus in healthy elderly subjects. Such seemingly contradictory findings highlight the importance to further evaluate the clinical effects of mTOR manipulation, including its potential role in treatment of COVID-19 infection. mTOR blockade may extend healthy lifespan by abrogating inflammation induced by viral infections and autoimmunity. This review provides a mechanistic assessment of mTOR pathway activation in lineage specification within the adaptive and innate immune systems and its role in health and autoimmunity. We then discuss some of the recent experimental and clinical discoveries implicating mTOR in viral pathogensis and aging.


Purification and properties of a mitochondrial NAD+ glycohydrolase.
A 60- to 70-fold purification of an NAD+ glycohydrolase from the inner membrane of rat liver mitochondria to apparent homogeneity on sodium dodecyl sulfate (SDS)-polyacrylamide slab gel is described. The minimum molecular weight of the enzyme on polyacrylamide gels in the presence of SDS is around 62,000. The enzyme splits NAD+ to ADP-ribose and, presumably, nicotinamide. No phosphatase or phosphodiesterase activity is detected in the purified enzyme preparation. The enzyme shows high activity with NAD+ and moderate activity with NADP+ as substrates NAD(P)Hs are poor substrates. ATP and nicotinamide inhibit the enzyme. A possible participation of the enzyme in the mechanism of calcium release from rat liver mitochondria is discussed.


Visual impairment in children.
This article examines the paediatric ophthalmic nurse's role in treating and managing several common ocular conditions.


Stress-induced cardiomyopathy (Tako-Tsubo) in a premenopausal woman: A case report from Saudi Arabia.
Tako-Tsubo cardiomyopathy (TTC) is a nonischemic cardiomyopathy characterized by reversible left ventricular dysfunction that is seen predominantly in postmenopausal women (>80%). The syndrome has symptoms that are similar to acute myocardial infarction, such as electrocardiogram changes (ST-segment elevation and subsequent giant T wave inversion) and abnormal cardiac enzymes. The clinical prognosis is usually benign. This article reports the first case of a TTC in a premenopausal Saudi woman. Early diagnosis of TTC excludes the use of stents, thrombolytics, and long-term coronary heart disease medications.


Negative regulation of the hepatitis B virus pre-S1 promoter by internal DNA sequences.
Expression of the surface antigen gene (S gene) of hepatitis B virus is directed by two distinct promoter elements with markedly different activities. The upstream (pre-S1) promoter produces a 2.4-kb transcript at very low levels while the downstream (pre-S2) promoter produces an approximately 2.1-kb transcript in relative abundance. We have constructed a series of internal deletion mutants to analyze differential regulation of the two S gene promoters. We show here that expression directed by the pre-S1 promoter is negatively regulated by DNA sequences containing the downstream pre-S2 promoter region. Nuclear run-on analysis indicates this down-regulation to be at the level of transcription. Furthermore, promoter repression does not appear to be due to products of the S gene region. Deletion mutagenesis studies have permitted the localization of a 61-bp region that may be involved in the apparent down-regulation of the pre-S1 promoter. These results suggest the use of an unusual regulatory mechanism by a dipromoter gene in which an active internal promoter may preclude efficient use of an upstream promoter.


Multiple myeloma cells promote migration of bone marrow mesenchymal stem cells by altering their translation initiation.
The role of the bone marrow microenvironment in multiple myeloma pathogenesis and progression is well recognized. Indeed, we have shown that coculture of bone marrow mesenchymal stem cells from normal donors and multiple myeloma cells comodulated translation initiation. Here, we characterized the timeline of mesenchymal stem cells conditioning by multiple myeloma cells, the persistence of this effect, and the consequences on cell phenotype. Normal donor mesenchymal stem cells were cocultured with multiple myeloma cell lines (U266, ARP1) (multiple myeloma-conditioned mesenchymal stem cells) (1.5 h,12 h, 24 h, 48 h, and 3 d) and were assayed for translation initiation status (eukaryotic translation initiation factor 4E; eukaryotic translation initiation factor 4G; regulators: mechanistic target of rapamycin, MNK, 4EBP; targets: SMAD family 5, nuclear factor κB, cyclin D1, hypoxia inducible factor 1, c-Myc) (immunoblotting) and migration (scratch assay, inhibitors). Involvement of mitogen-activated protein kinases in mesenchymal stem cell conditioning and altered migration was also tested (immunoblotting, inhibitors). Multiple myeloma-conditioned mesenchymal stem cells were recultured alone (1-7 d) and were assayed for translation initiation (immunoblotting). Quantitative polymerase chain reaction of extracted ribonucleic acid was tested for microRNAs levels. Mitogen-activated protein kinases were activated within 1.5 h of coculture and were responsible for multiple myeloma-conditioned mesenchymal stem cell translation initiation status (an increase of >200%, P < 0.05) and elevated migration (16 h, an increase of >400%, P < 0.05). The bone marrow mesenchymal stem cells conditioned by multiple myeloma cells were reversible after only 1 d of multiple myeloma-conditioned mesenchymal stem cell culture alone. Decreased expression of microRNA-199b and microRNA-125a (an increase of <140%, P < 0.05) in multiple myeloma-conditioned mesenchymal stem cells supported elevated migration. The time- and proximity-dependent conditioning of normal donor mesenchymal stem cells in our model points to a dynamic interaction between multiple myeloma cells and the bone marrow niche, which causes profound changes in the nonmalignant bone marrow constituents. Future studies are warranted to identify clinically relevant means of blocking this crosstalk and improving multiple myeloma therapy.


Salmonella typhi: from a human pathogen to a vaccine vector.
Salmonella (S.) typhi is an important intracellular pathogen. Among the more than 2,300 closely-related Salmonella serovars bacteria recognized, S. typhi is the only one that is pathogenic exclusively for humans, in whom it causes typhoid or enteric fever. The pathogen has been around for many years and many studies have been done in an effort to combat it. Molecular and biologic features of S. typhi and host factors and immune responses involved in Salmonella invasion have been extensively studies. Vaccines that have been developed most notably are Vi polysaccharide and Ty21a. However, as the results show, there is still a long way to go. It is also shown that multi-drug resistance has occurred to the few available antibiotics. More and more studies have shown that Salmonella can be used as a vaccine vector carrying antigens of other pathogens. This has been promising in that the immune system can be elicited in response to both the Salmonella bacteria and the antigen of the pathogen in question. This review aims to highlight some of the milestones attained in the fight against the disease from the time S. typhi was seen as a pathogen causing typhoid fever to the use of Salmonella as a vaccine vector.


Impact of medium volume and oxygen concentration in the incubator on pericellular oxygen concentration and differentiation of murine chondrogenic cell culture.
Previous studies have demonstrated that oxygen environment is an important determinate factor of cell phenotypes and differentiation, although factors which affect pericellular oxygen concentration (POC) in murine chondrogenic cell culture remain unidentified. Oxygen concentrations in vivo were measured in rabbit musculoskeletal tissues, which were by far hypoxic compared to 20% O(2) (ranging from 2.29 ± 1.16 to 4.36 ± 0.51%). Oxygen concentrations in murine chondrogenic cell (C3H10T1/2) culture medium were monitored in different oxygen concentrations (20% or 5%) in the incubator and in different medium volumes (3,700 or 7,400 μl) within 25-cm(2) flasks. Chondrogenic differentiation was assessed by glycosaminoglycan production with quantitative evaluation of Alcian blue staining in 12-well culture dishes. Expression of chondrogenic genes, aggrecan, and type II collagen α1, was examined by quantitative real-time polymerase chain reaction. Oxygen concentrations in medium decreased accordingly with the depth from medium surface, and POC at Day 6 was 18.99 ± 0.81% in 3,700-μl medium (1,480-μm depth) and 13.26 ± 0.23% in 7,400-μl medium (2,960-μm depth) at 20% O(2) in the incubator, which was 4.96 ± 0.08% (1,480-μm depth) and 2.83 ± 0.42% (2,960-μm depth) at 5% O(2), respectively. The differences of POC compared by medium volume were statistically significant (p = 0.0003 at 20% and p = 0.001 at 5%). Glycosaminoglycan production and aggrecan gene expression were most promoted when cultured in moderately low POC, 1,000 μl (2,960-μm depth) at 20% O(2) and 500 μl (1,480-μm depth) at 5% O(2) in 12-well culture dishes. We demonstrate that medium volume and oxygen concentration in the incubator affect not only POC but also chondrogenic differentiation.