Membrane cycling between the ER and Golgi apparatus and its role in biosynthetic transport.
Membrane traffic between the ER and Golgi is now recognized as a carefully regulated process controlled by distinct anterograde (to the Golgi) and retrograde (to the ER) pathways. These pathways link two organelles with different morphologies, structures, and localizations within the cell. The ER, which is involved in multiple cellular functions including protein biosynthesis and folding, extends to the cell periphery and forms a dynamic tubule reticulum. By contrast, the Golgi apparatus, which functions in membrane sorting and recycling events, is localized at the center of the cell near the MTOC and is comprised of compact cisternal units. The required transport into the Golgi apparatus of newly synthesized proteins exported from the ER offers a twofold advantage to the cell. First, the rate of movement of membrane and protein through the biosynthetic pathway can be controlled by the selective use of a recycling pathway. Second, membrane moving through the biosynthetic pathway enters a structure specialized for sorting of membrane to different final destinations in the cell Control of biosynthetic transport within the ER/Golgi system involves the utilization of two alternative transport pathways: anterograde (ER to Golgi) and retrograde (Golgi to ER). These two pathways share a common regulatory system involving membrane assembly/disassembly of cytosolic coatomer proteins. Thus, conditions that favor irreversible coatomer binding (i.e., GTP gamma S) inhibit retrograde transport while producing anterograde transport intermediates. Conditions that prevent coatomer binding (i.e., BFA) inhibit anterograde transport and enhance retrograde transport. The underlying biochemical machinery that normally balances anterograde and retrograde membrane traffic between the ER and Golgi is only just beginning to be understood. Any model to explain this system, however, must account for the morphologic characteristics of the membranes involved. Whereas anterograde traffic involves discontinuous "coated" structures moving from peripheral sites in the ER toward the central Golgi, retrograde traffic utilizes continuous "noncoated" tubule structures that move from a central site (i.e., the CGN) to the peripheral ER (see Figure 3). Such a system maximizes volume transport (utilizing vacuolar structures) in the anterograde direction and membrane transport (utilizing tubules) in the retrograde direction. It is therefore ideal for sorting of bulk flow lumenal contents from recycling membrane early in the biosynthetic pathway.


A tripartite organization of the urbilaterian brain: developmental genetic evidence from Drosophila.
Developmental genetic studies suggest that the embryonic vertebrate brain has a tripartite ground plan consisting of a forebrain/midbrain, a hindbrain, and an intervening midbrain/hindbrain boundary region, which are characterized by the specific expression of the Otx, Hox and Pax-2/5/8 genes. Recent studies in Drosophila reveal similarities in the expression and function of these genes in patterning the embryonic brains of flies and vertebrates. Thus, in Drosophila, as is vertebrates, a Pax2/5/8 domain is located between an anterior otd/Otx2 region and a posterior Hox region of the embryonic brain. Moreover, in Drosophila, as in vertebrates, this Pax2/5/8 domain is located at the interface of the otd/Otx2 domain and a posterior unplugged/Gbx2 domain. Furthermore, in Drosophila, as in vertebrates, inactivation of otd/Otx2 or of unplugged/Gbx2 results in a comparable mispositioning or loss of orthologous gene expression domains in the embryonic brain. These developmental genetic similarities suggest that the tripartite ground plan, which characterizes the developing vertebrate brain, is also at the basis of the developing insect brain. This, in turn, implies that a tripartite organization of the embryonic brain may characterize all extant bilaterians, and thus may already have been established in the last common urbilaterian ancestor of all bilaterians.


Cell-to-cell variability in troponin I phosphorylation in a porcine model of pacing-induced heart failure.
We tested the hypothesis that myocardial contractile protein phosphorylation and the Ca(2+) sensitivity of force production are dysregulated in a porcine model of pacing-induced heart failure (HF). The level of protein kinase A (PKA)-dependent cardiac troponin I (TnI) phosphorylation was lower in the myocardium surrounding the pacing electrode (pacing site) of the failing left ventricle (LV) than in the controls. Immunohistochemical assays of the LV pacing site pointed to isolated clusters of cardiomyocytes exhibiting a reduced level of phosphorylated TnI. Flow cytometry on isolated and permeabilized cardiomyocytes revealed a significantly larger cell-to-cell variation in the level of TnI phosphorylation of the LV pacing site than in the opposite region in HF or in either region in the controls: the interquartile range (IQR) on the distribution histogram of relative TnI phosphorylation was wider at the pacing site (IQR = 0.53) than that at the remote site of HF (IQR = 0.42; P = 0.0047) or that of the free wall of the control animals (IQR = 0.36; P = 0.0093). Additionally, the Ca(2+) sensitivities of isometric force production were higher and appeared to be more variable in single permeabilized cardiomyocytes from the HF pacing site than in the healthy myocardium. In conclusion, the level of PKA-dependent TnI phosphorylation and the Ca(2+) sensitivity of force production exhibited a high cell-to-cell variability at the LV pacing site, possibly explaining the abnormalities of the regional myocardial contractile function in a porcine model of pacing-induced HF.


Tracheotomy does not affect reducing sedation requirements of patients in intensive care--a retrospective study.
Translaryngeal intubated and ventilated patients often need sedation to treat anxiety, agitation and/or pain. Current opinion is that tracheotomy reduces sedation requirements. We determined sedation needs before and after tracheotomy of intubated and mechanically ventilated patients. We performed a retrospective analysis of the use of morphine, midazolam and propofol in patients before and after tracheotomy. Of 1,788 patients admitted to our intensive care unit during the study period, 129 (7%) were tracheotomized. After the exclusion of patients who received a tracheotomy before or at the day of admittance, 117 patients were left for analysis. The daily dose (DD; the amount of sedatives for each day) divided by the mean daily dose (MDD; the mean amount of sedatives per day for the study period) in the week before and the week after tracheotomy was 1.07 +/- 0.93 DD/MDD versus 0.30 +/- 0.65 for morphine, 0.84 +/- 1.03 versus 0.11 +/- 0.46 for midazolam, and 0.62 +/- 1.05 versus 0.15 +/- 0.45 for propofol (p < 0.01). However, when we focused on a shorter time interval (two days before and after tracheotomy), there were no differences in prescribed doses of morphine and midazolam. Studying the course in DD/MDD from seven days before the placement of tracheotomy, we found a significant decline in dosage. From day -7 to day -1, morphine dosage (DD/MDD) declined by 3.34 (95% confidence interval -1.61 to -6.24), midazolam dosage by 2.95 (-1.49 to -5.29) and propofol dosage by 1.05 (-0.41 to -2.01). After tracheotomy, no further decrease in DD/MDD was observed and the dosage remained stable for all sedatives. Patients in the non-surgical and acute surgical groups received higher dosages of midazolam than patients in the elective surgical group. Time until tracheotomy did not influence sedation requirements. In addition, there was no significant difference in sedation between different patient groups. In our intensive care unit, sedation requirements were not further reduced after tracheotomy. Sedation requirements were already sharply declining before tracheotomy was performed.


A circular RNA map for human induced pluripotent stem cells of foetal origin.
Adult skin fibroblasts represent the most common starting cell type used to generate human induced pluripotent stem cells (F-hiPSC) for clinical studies. Yet, a foetal source would offer unique advantages, primarily the absence of accumulated somatic mutations. Herein, we generated hiPSC from cord blood multipotent mesenchymal stromal cells (MSC-hiPSC) and compared them with F-hiPSC. Assessment of the full activation of the pluripotency gene regulatory network (PGRN) focused on circular RNA (circRNA), recently proposed to participate in the control of pluripotency. Reprogramming was achieved by a footprint-free strategy. Self-renewal and pluripotency of cord blood MSC-hiPSC were investigated in vitro and in vivo, compared to parental MSC, to embryonic stem cells and to F-hiPSC. High-throughput array-based approaches and bioinformatics analyses were applied to address the PGRN. Cord blood MSC-hiPSC successfully acquired a complete pluripotent identity. Functional comparison with F-hiPSC showed no differences in terms of i) generation of mesenchymal-like derivatives, ii) their subsequent adipogenic, osteogenic and chondrogenic commitment, and iii) their hematopoietic support ability. At the transcriptional level, specific subsets of mRNA, miRNA and circRNA (n = 4,429) were evidenced, casting a further layer of complexity on the PGRN regulatory crosstalk. A circRNA map of transcripts associated to naïve and primed pluripotency is provided for hiPSC of clinical-grade foetal origin, offering insights on still unreported regulatory circuits of the PGRN to consider for the optimization and development of efficient differentiation protocols for clinical translation. This research was funded by Ricerca Corrente 2012-2018 by the Italian Ministry of Health.


The influence of a diabetes awareness program on diabetes knowledge, risk perception, and practices among university students.
To compare diabetes mellitus (DM) knowledge, risk perception, and health practices before and after a mass awareness program among Princess Nourah bint Abdulrahman University students, Riyadh, Saudi Arabia. A DM awareness program was conducted between November 2020 and February 2021 in three phases (total 13 weeks). The program comprised educational messages, short videos, virtual lectures, and peer-to-peer education. A total of 558 participants (before group, n = 360; after group, n = 198) were included in the study. Majority of the students had normal body mass index (BMI). The knowledge scores were significantly higher after the program (median 16, quartile range 13-18) than before (median 14, quartile range 12-16) (p < 0.001). No significant differences were observed in risk perception between the two groups (p > 0.05). In addition, increased moderate practice score and decreased poor practice score (p < 0.001) were observed. Factors influencing DM knowledge were higher education and awareness. Factors influencing health practices were higher BMI and knowledge after the program. DM awareness program improved DM knowledge and health practices among university students.


The smooth microsomal fractions from Aspergillus oryzae were shown to carry out transfer of mannose from GDP-mannose to endogenous proteins acceptors. Data also are presented which support that among the subcellular fractions, only the microsomes are active in the mannose transfer reaction. All of the radioactive material chromatographs with mannose, following acid hydrolysis of microsomal fractions.


[Ethical reflection in structures, a compass in the crisis].
Morality, ethics, ethical approach are words that are often used in our daily lives as health care providers. What meaning do they have? How important are they? If ethics is an individual notion, the question arises of its application in our accompaniment.


Recombinant human MFG-E8 attenuates cerebral ischemic injury: its role in anti-inflammation and anti-apoptosis.
Excessive inflammation and apoptosis contribute to the pathogenesis of ischemic stroke. MFG-E8 is a 66-kDa glycoprotein that has shown tissue protection in various models of organ injury. However, the potential role of MFG-E8 in cerebral ischemia has not been investigated. We found that levels of MFG-E8 protein in the brain were reduced at 24 h after cerebral ischemia. To assess the potential role of MFG-E8 in cerebral ischemia, adult male Sprague-Dawley rats were subjected to permanent middle cerebral artery occlusion (MCAO). At 1 h post-stroke onset, an intravenous administration of 1 ml saline as vehicle or 160 μg/kg BW recombinant human MFG-E8 (rhMFG-E8) as treatment was given. The optimal dose of rhMFG-E8 was obtained from previous dose-response organ protection in rat sepsis studies. Neurological scores were determined at 24 h and 48 h post-MCAO. Rats were sacrificed thereafter and brains rapidly removed and analyzed for infarct size, histopathology, and markers of inflammation and apoptosis. Compared with saline vehicle, rhMFG-E8 treatment led to significant decreases in sensorimotor and vestibulomotor deficits, and infarct size at 24 h and 48 h post-MCAO. Measures associated with improved outcome included reduced microglial inflammatory cytokine secretion, adhesion molecules and neutrophil influx, cleaved caspase-3, and upregulation of peroxisome proliferator activated receptor-γ (PPAR-γ), and Bcl-2/Bax ratio leading to decreased apoptosis. Thus, rhMFG-E8 treatment is neuroprotective against cerebral ischemia through suppression of inflammation and apoptosis. This article is part of a Special Issue entitled 'Post-Traumatic Stress Disorder'.


Immune activation and depression in women with rheumatoid arthritis.
We examined markers of immune activation during periods of stress and depressive symptoms in 45 female patients with rheumatoid arthritis (RA) in comparison to 106 controls with no autoimmune disease. Depressive symptoms were recorded, clinician ratings of disease activity were made, and blood was drawn for RA patients and controls at baseline and during a designated stressful week. Counts of T cell subpopulations revealed significant differences between RA and control groups in proportions of CD8 and CD4 cells, with higher CD4 and lower CD8 counts for the RA participants. Significant depression by diagnosis interactions were found, revealing greater CD4 activation among RA patients who were depressed in comparison to other groups. Only marginally significant effects of stress were found on T cell counts. Interleukin 6 (IL-6) concentrations also differentiated groups, with the highest levels of IL-6 observed for depressed RA patients under stress. These findings provide new evidence that psychosocial factors play a significant role in autoimmune processes that underlie RA.


Fractionation of rosemary (Rosmarinus officinalis L.) essential oil using vacuum fractional distillation.
Rosemary essential oil has many compounds suitable for use in the food, cosmetics and pharmaceuticals industries. This study aimed to evaluate vacuum fractional distillation to separate and purify components of rosemary oil, also to observe the distillation patterns. The light terpenes (α-pinene, myrcene, camphene) were removed at the top of the column, while the oxygen containing components (verbenone, borneol, linalool, geraniol) remained at the bottom. The concentrations of some minor compounds increased more than ten times (borneol, geraniol) compared with the raw oil. There was no evidence of thermal degradation in any of the experiments. Therefore, vacuum fractional distillation may be an interesting process to upgrade rosemary essential oil.


DNA probes bind non-specifically to eosinophils during in situ hybridization: carbol chromotrope blocks binding to eosinophils but does not inhibit hybridization to specific nucleotide sequences.
Eosinophils were found to bind radiolabelled DNA probes non-specifically during in situ hybridization. Pretreatment of cells with carbol chromotrope blocked non-specific binding without interfering with the recognition of specific nucleotide sequences.


Calcitonin gene-related peptide-receptor component protein expression in the uterine cervix, lumbosacral spinal cord, and dorsal root ganglia.
The neuropeptide calcitonin gene-related peptide (CGRP) may play a role in neurogenic inflammation, tissue remodeling of the uterine cervix, promoting vasodilation, parturition, and processing of sensory information in the spinal cord. CGRP-immunoreactive nerves of the cervix and spinal cord have been studied but cellular identification of the CGRP receptor has received little attention. CGRP-receptor component protein (CGRP-RCP) is a small protein associated with the CGRP receptor; thus, immunostaining for the CGRP-RCP can be used to identify sites of the CGRP receptor. We determined sites of CGRP-RCP immunoreactivity relative to the presence of CGRP-ir nerve fibers in the female rat uterine cervix, spinal cord, and dorsal root ganglia. CGRP-RCP immunoreactivity was expressed in the dorsal horn of the spinal cord, venules of the uterine cervix, and perikarya of sensory neurons in dorsal root ganglia. CGRP-immunoreactive fibers were adjacent to CGRP-RCP-immunoreactive vessels in the cervix and among CGRP-RCP-immunoreactive structures in the dorsal horn of the spinal cord. This suggests CGRP-RCP is associated with structures innervated by CGRP nerves and these interactions may be changed in tissues in response to an appropriate stimulus.


Synthesis, spectroscopic and structural characterization, and biological activity of aquachloro (pyridoxal thiosemicarbazone) copper(II) chloride.
The synthesis, spectroscopic studies, x-ray crystal structure, and biological properties of the complex [Cu(H2L)(OH2)Cl]Cl (1) (H2L = pyridoxal thiosemicarbazone) are reported. The compound crystallizes in space group P2(1)/n, a = 12.128(2), b = 9.096(2), c = 13.592(2) A, beta = 108.65(2) degrees, U = 1420.7 A3, and Z = 4. The molecular structure consists of discrete cations [Cu(H2L)(OH2)Cl]+ and Cl- anions. Each copper atom is in an approximately square pyramidal environment involving the phenolic oxygen, the imine nitrogen, the sulphur, and a water oxygen in the equatorial positions, while a chlorine atom occupies the axial position. The structure of this complex is compared to that of the dimeric [(Cu(HL)(OH2))2]Cl2.2H2O (2) obtained under different experimental conditions, to that of a Co(III) complex with the same ligand [Co(HL)L].4.5H2O (3) and to that of the free ligand H2L, especially in relation to its biological activity. Compounds 1 and 2 have not antiviral action in vitro with respect to RNA viruses, show an inductive effect on Friend erythroleukemia cells (FLC), erythroid differentiation and a suppressive effect regarding FLC proliferation. Complex 3 and the free ligand do not have biological activity.


Metrnl ameliorates diabetic cardiomyopathy via inactivation of cGAS/STING signaling dependent on LKB1/AMPK/ULK1-mediated autophagy.
Meteorin-like hormone (Metrnl) is ubiquitously expressed in skeletal muscle, heart, and adipose with beneficial roles in obesity, insulin resistance, and inflammation. Metrnl is found to protect against cardiac hypertrophy and doxorubicin-induced cardiotoxicity. However, its role in diabetic cardiomyopathy (DCM) is undefined. We aimed to elucidate the potential roles of Metrnl in DCM. Gain- andloss-of-function experimentswere utilized to determine the roles of Metrnl in the pathological processes of DCM. We found that plasma Metrnl levels, myocardial Metrnl protein and mRNA expressions were significantly downregulated in both streptozotocin (STZ)-induced (T1D) mice and leptin receptor deficiency (db/db) (T2D) mice. Cardiac-specific overexpression (OE) of Metrnl markedly ameliorated cardiac injury and dysfunction in both T1D and T2D mice. In sharp contrast, specific deletion of Metrnl in the heart had the opposite phenotypes. In parallel, Metrnl OE ameliorated, whereas Metrnl downregulation exacerbated high glucose (HG)-elicited hypertrophy, apoptosis and oxidative damage in primary neonatal rat cardiomyocytes. Antibody-induced blockade of Metrnl eliminated the effects of benefits of Metrnl in vitro and in vivo. Mechanistically, Metrnl activated the autophagy pathway and inhibited the cGAS/STING signaling in a LKB1/AMPK/ULK1-dependent mechanism in cardiomyocytes. Besides, Metrnl-induced ULK1 phosphorylation facilitated the dephosphorylation and mitochondrial translocation of STING where it interacted with tumor necrosis factor receptor-associated factor 2 (TRAF2), a scaffold protein and E3 ubiquitin ligase that was responsible for ubiquitination and degradation of STING, rendering cardiomyocytes sensitive to autophagy activation. Thus, Metrnl may be an attractive therapeutic target or regimen for treating DCM.


Development and Validation of a Highly Sensitive Liquid Chromatography-Tandem Mass Spectrometry Technique to Determine Levamisole in Plasma and Saliva.
Levamisole is used as a steroid-sparing drug for the treatment of frequently relapsing or steroid-dependent idiopathic nephrotic syndrome in children. As part of a large multicentre randomized controlled trial with levamisole, pharmacokinetic and pharmacodynamic parameters of levamisole in children with idiopathic nephrotic syndrome were investigated, as well as the feasibility of using saliva as an alternative and patient-friendly matrix for determining levamisole concentrations. In this study, the authors presented the development and validation of a highly sensitive method for determining levamisole in plasma and saliva using liquid chromatography-tandem mass spectrometry (LC-MS/MS). In 100 μL samples, proteins were precipitated with 750 μL acetonitrile/methanol 420:80 (v/v) with levamisole-D5 as an internal standard. Calibration standards were prepared over a range of 0.1 ng/mL-50 ng/mL. To determine ultrafiltration efficiency, the ultrafiltrate was obtained by centrifuging blank plasma samples over the filter. Both filtered and nonfiltered samples were analyzed. For plasma, accuracy and within-run and between-run imprecision were between 95.0% and 100% and <14.5%, respectively, and for saliva, between 100.9% and 107.5%, and <13.3%. No significant matrix effects were observed. Samples were stable at benchtop for 24 hours and -80°C, for at least 14 months (stability experiments ongoing). The ultrafiltration efficiency of unbound concentrations in plasma was lower than 85% (58.9%) but stable, and, therefore, the observed concentration should be corrected. Based on observations, the developed measure can determine levamisole concentrations in participant saliva samples.


Surgical repair for abdominal aortic aneurysm concomitant with iliac arterial disease using InterGard™ Quadrifurcated.
InterGard™ Quadrifurcated [InterVascular S.A. (MAQUET Cardiovascular), La Ciotat, France], a knitted quadrifurcated prosthetic graft, is useful to simplify the procedures for patients with abdominal aortic aneurysm concomitant with iliac arterial disease. From March 2008 to April 2010, 59 patients underwent abdominal aortic aneurysm repair in our department. InterGard™ Quadrifurcated was used in 22 patients (37.3%). All patients were male with a mean age of 72.1 ± 12.1 years (range from 45 to 90 years). Four were emergency cases. Nineteen patients had common iliac arterial lesions, and 19 patients had internal iliac arterial lesions. In addition to abdominal aortic aneurysm repair, reconstruction of bilateral internal iliac arteries was performed in 4 patients, reconstruction of unilateral internal iliac artery in 15 (right 6, left 9), and internal mesenteric artery in 13. There was one hospital death due to nonocclusive mesenteric infarction. There were two complications, which were intestinal perforation and intestinal obstruction. All of these three cases were emergency cases. Computed tomography scan 1 week after operation showed that all reconstructed grafts were patent. Although patients with abdominal aortic aneurysm concomitant with iliac arterial disease had many preoperative comorbidities, surgical results with InterGard™ Quadrifurcated were satisfactory. It is useful to simplify the procedures for these patients.


The role of HLA antibodies in allogeneic SCT: is the 'type-and-screen' strategy necessary not only for blood type but also for HLA?
The role of HLA antibodies in SCT has drawn increasing attention because of the significantly increased number of patients who receive HLA-mismatched SCT, including cord blood transplantation, haploidentical SCT and unrelated SCT. Technical advancements in the methods of HLA Ab testing have realized rapid, accurate and objective identification, as well as quantification of specific HLA antibodies. Recent clinical studies have suggested that the presence of donor-specific HLA antibodies (DSA) in patients is associated with graft failure in HLA-mismatched SCT when the above-listed stem cell sources are used and results in different impacts. Of note, most of the 'HLA-matched' unrelated SCT actually involve HLA mismatches in HLA-DP and the presence of antibodies against this locus has been reported to be associated with graft failure. Thus, HLA Ab should be examined as a work-up for all patients who undergo SCT from 'alternative donors.' The simplest route for preventing HLA Ab-mediated graft failure in Ab-positive patients is to avoid donors who possess the target Ag of HLA antibodies. If SCT from such donors must be performed, treatment for DSA before SCT may improve the chances of successful donor engraftment.


Exercise urotensin II dynamics in myocardial infarction survivors with and without hypertension.
Hypertension is diagnosed in approximately 50% of patients with acute myocardial infarction. Urotensin II (U-II) - a potent vasoactive peptide shown to be elevated in hypertensive subjects, can contribute to negative myocardial remodelling and development of left ventricular failure. Data concerning U-II activity under exercise conditions and its influence on blood pressure in patients after myocardial infarction is scant. Therefore we sought to determine U-II dynamics during exercise in myocardial infarction survivors with and without hypertension. Forty patients with acute myocardial infarction treated with successful primary coronary angioplasty, after four weeks of uneventful and symptom-free period following initial hospitalization underwent treadmill exercise test. U-II plasma concentration was measured before and shortly after the exercise. Hypertension was diagnosed in 17 (42.5%) patients. We found no significant differences between normotensive and hypertensive subjects except higher smoking rate and lower calcium channel blockers prescription in normotensive patients. Both systolic and diastolic blood pressure were comparable between study groups before exercise. After exercise we observed higher systolic blood pressure in hypertensive subjects (169.06 +/- 30.23 vs. 150.0 +/- 18.97 mm Hg; p < 0.05). U-II concentration showed no significant difference in pretest sampling (54.93 +/- 38.11 vs. 73.97 +/- 48.52 ng/ml; p = NS). After exercise we noted significantly higher peptide level in hypertensive patients (63.32 +/- 36.11 vs. 98.03 +/- 40.47 ng/ml; p = 0.01). The present study is the first one to show differences in U-II concentration exercise dynamics in hypertensive and normotensive myocardial infarction survivors. It sheds additional light on hypertension pathophysiology in myocardial infarction patients, and thus identifies a novel, potentially relevant, target for future therapeutic interventions.


Endoscopic Lumbar Interbody Fusion.
Spine surgeons have recently developed more advanced and less invasive techniques. One significant example of recent surgical innovation is the advent of endoscopic-assisted spine surgery. Endoscopic lumbar interbody fusions are increasingly used to treat lumbar degenerative disease in a minimally invasive approach that minimizes pain and maximizes outcomes. Numerous technical refinements to ultraminimally invasive approaches have occurred since their initial use, which has resulted in substantial clinical benefit for patients that remains stable over time. Serious complications can occur and thus, the spine surgeon adopting endoscopic techniques must be aware of the early learning curve.


Injectable pharmacotherapy for opioid use disorders (IPOD).
Despite the growing prevalence of opioid use among offenders, pharmacotherapy remains an underused treatment approach in correctional settings. The aim of this 4-year trial is to assess the clinical utility, effectiveness, and cost implications of extended-release naltrexone (XR-NTX, Vivitrol®; Alkermes Inc.) alone and in conjunction with patient navigation for jail inmates with opioid use disorder (OUD). Opioid-dependent inmates will be randomly assigned to one of three treatment conditions before being released to the community to include: 1) XR-NTX only; 2) XR-NTX plus patient navigation (PN), and 3) enhanced treatment-as-usual (ETAU) with drug education and a community treatment referral. Before release from jail, participants in the XR-NTX and XR-NTX plus PN conditions will receive their first XR-NTX injection. Those in the XR-NTX plus PN condition also will meet with a patient navigator. Participants in both XR-NTX conditions will be scheduled for medical management sessions twice monthly for months 1-3, monthly medical management sessions for months 4-6, with monthly injections for 5months post-release (which, given the pre-release injection, results in a 6-month medication phase). Follow-up data collection will occur at 1, 3, 6, and 12months post release. We discuss the study's rationale, aims, methods, and anticipated findings. The primary outcome is the presence of a DSM 5 OUD diagnosis 1year after randomization (6months after the end of the active treatment phase). We hypothesize that providing XR-NTX prior to release from jail will be particularly beneficial for this extremely high-risk population by reducing opioid use, associated criminal behavior, and injection-related disease risk. ClinicalTrials.Gov: NCT02110264.


Artifact weathering, anthropogenic microparticles and lead contamination in urban soils at former demolition sites, Detroit, Michigan.
A chronological sequence of urban soils 3-92 years old was studied to determine the effects of time on morphogenesis, artifact weathering, and the geochemical partitioning of Pb. Key chronofunctions determined are an increase in ˆA horizon Development Index (defined herein based on soil color) and water-soluble Pb, and a decrease in pH and C/N, with increasing soil age. Key artifact weathering reactions are: 1) portlandite in mortar altered to calcite, 2) ferrite in wrought-iron altered to ferrihydrite and goethite, and 3) carbonaceous materials altered to water-soluble organic substances. Mortar and wrought-iron were found to be Pb-bearing, but weather to produce immobilizing agents. Hence, they are both a source and a sink for Pb. The origin and mobilization of water-soluble Pb is complex and probably includes microbial extracellular polymeric substances, biodegraded soil organic matter, and solubilized organic substances derived from carbonaceous anthropogenic microparticles (soot, char and coal-related wastes).


The prevalence of coeliac disease-associated human leukocyte antigens in South African transplant donors and recipients.
Coeliac disease (CD) is an autoimmune condition occurring in genetically predisposed individuals exposed to an environmental trigger. The human leukocyte antigen (HLA) haplotypes HLA-DQ2.5 and HLA-DQ8 have the strongest association with CD, and 90 - 95% of CD patients bear these haplotypes. The susceptibility of the South African (SA) population to CD has not been studied previously. To describe the genetic propensity of the SA population to CD. The South African National Blood Service database was used to analyse the prevalence of HLA-DQ2.5 and HLA-DQ8 in potential donors and recipients of organ transplants. Self-reported ethnic group was used to estimate the prevalence among different population groups. The overall prevalence of HLA-DQ2.5 and HLA-DQ8 was 19.8%. The prevalence was lower in black participants (15.9%) than in whites (28.6%). Coloured (22.0%) and Indian (17.4%) participants had an intermediate prevalence. There was no significant difference between potential transplant donors and recipients. The prevalence of HLA-DQ2.5 and HLA-DQ8 differed among SA study participants of different ethnicities. However, the notion that CD does not occur in black South Africans owing to lack of a genetic predisposition is incorrect.


The effects of reformulation: improved therapeutic index.
Antimuscarinic agents are the treatment of choice for overactive bladder syndrome. Due to the development of novel delivery systems, extended-release formulations of oxybutynin, tolterodine, and trospium chloride are now available. In addition to the convenience of once-daily dosing, the new formulations of these commonly prescribed agents have improved their therapeutic index, striking a better balance between efficacy and tolerability.


Familial influences on Mexican American and Euro-American preadolescent boys' and girls' body dissatisfaction.
This study examines multiple influences on preadolescent boys' and girls' body dissatisfaction over time. Mexican American and Euro-American preadolescents (n=105) completed measures addressing their body dissatisfaction. In addition, preadolescents' parents completed a measure assessing their own body dissatisfaction. Results indicate that boys' body dissatisfaction was related to their body size (body mass index, BMI) and their fathers' own body dissatisfaction; girls' body dissatisfaction was related primarily to their own body size. Findings are discussed in terms of the importance of understanding boys' increasing susceptibility to body dissatisfaction and the role fathers play in shaping boys' feelings about their bodies as they approach adolescence.


Tissue engineering of human cartilage and osteochondral composites using recirculation bioreactors.
Chondrocytes isolated from human foetal epiphyseal cartilage were seeded dynamically into polyglycolic acid (PGA) scaffolds and cultured in recirculation column bioreactors to produce tissue-engineered cartilage. Several culture techniques with the potential to provide endogenous growth factors and other conditions beneficial for de novo cartilage synthesis were investigated. Osteochondral composite constructs were generated by seeding separate PGA scaffolds with either foetal chondrocytes or foetal osteoblasts then suturing the scaffolds together before bioreactor cultivation. This type of co-culture system provided direct contact between the tissue-engineered cartilage and developing tissue-engineered bone and yielded significant improvements in cartilage quality. In the cartilage section of the composites, the concentrations of glycosaminoglycan (GAG) and total collagen were increased by 55% and 2.5-fold, respectively, compared with control cartilage cultures, while levels of collagen type II were similar to those in the controls. The osteochondral composites were harvested from the bioreactors as single units with good integration between the cartilage and bone tissues. Only the cartilage layer contained GAG while only the bone layer was mineralised. In other experiments, co-culture of tissue-engineered cartilage with pieces of ex-vivo cartilage or ex-vivo bone did not improve the quality of the cartilage relative to control cultures. Addition of 10(-6) M diacerein to the culture medium also had no effect on the properties of engineered cartilage. This work demonstrates the beneficial effects of generating cartilage tissues in contact with developing bone. It also demonstrates the feasibility of producing composite osteochondral constructs for clinical application using recirculation column bioreactors.


Identification of two novel fibrinolytic enzymes from Bacillus subtilis QK02.
Two fibrinolytic enzymes (QK-1 and QK-2) purified from the supernatant of Bacillus subtilis QK02 culture broth had molecular masses of 42,000 Da and 28,000 Da, respectively. The first 20 amino acids of the N-terminal sequence are AQSVPYGISQ IKAPALHSQG. The deduced protein sequence and its restriction enzyme map of the enzyme QK-2 are different from those of other proteases. The enzyme QK-2 digested not only fibrin but also a subtilisin substrate, and PMSF inhibited its fibrinolytic and amidolytic activities completely; while QK-1 hydrolyzed fibrin and a plasmin substrate, and PMSF as well as aprotinin inhibited its fibrinolytic activity. These results indicated QK-1 was a plasmin-like serine protease and QK-2 a subtilisin family serine protease. Therefore, these enzymes were designated subtilisin QK. The sequence of a DNA fragment encoding subtilisin QK contained an open reading frame of 1149 base pairs encoding 106 amino acids for signal peptide and 257 amino acids for subtilisin QK, which is highly similar with that of a fibrinolytic enzyme, subtilisin NAT (identities 96.8%). Asp32, His64 and Ser221 in the amino acid sequence deduced from the QK gene are identical to the active site of nattokinase (NK) produced by B. subtilis natto.


Recent progress in the research of yam mucilage polysaccharides: Isolation, structure and bioactivities.
Yam (Dioscorea spp.), known as an edible and medicinal tuber crop in China, has been used historically for the treatment of diabetes, diarrhea, asthma, and other ailments in traditional Chinese medicine. Moreover, it has been consumed as starchy food for thousands of years in China. Modern phytochemistry and pharmacological experiments have been proved that non-starch polysaccharide is one of the main bioactive substances of yam. Many studies have been focused on the isolation and identification of polysaccharides and their bioactivities of Chinese yam. However, due to the difference in the variety of raw materials and the method of polysaccharides extracting, the structure and biological activity of the obtained polysaccharides also differ. It has been demonstrated that Chinese yam polysaccharide has various important biological activities, such as hypoglycemia, immunomodulatory, antioxidant, and antitumor activities. This paper is aimed at summarizing previous and current references of the isolation processes, structural features and bioactivities of yam polysaccharides. The review will serve as a useful reference material for further investigation and application of yam polysaccharides in functional foods and medicine fields.


The effect of oxygen tension on haemopoietic and fibroblast cell proliferation in vitro.
The effects of providing low oxygen tension in the gas phase of two different types of cell culture systems were investigated. The clonal growth of granulocyte-macrophage progenitor cells in an agar culture system was improved markedly by incubation within a low oxygen tension gas phase (48 mmHg--6.8%) instead of the conventional air (135 mmHg--19%), the effects being measured by increases in numbers of colony forming cells detected and in the colony sizes. The increased efficiency of colony formation was observed both with mouse and human marrow cells. A similar effect was observed in a liquid adherence culture system with primary cultures of foetal mouse fibroblasts both at clonal and higher cell densities.


Nanoscale metal-organic frameworks as key players in the context of drug delivery: evolution toward theranostic platforms.
Metal-organic frameworks (MOFs) have emerged as one of the most fascinating libraries of porous materials with a huge potential in very diverse application areas. In particular, the bioanalytical and biomedical fields have evolved tremendously due to the emergence of these hybrid inorganic-organic MOF-based materials. This is because these materials possess a series of key properties essential for bioapplications, such as minimal toxicity to living cells, intrinsic biodegradability, and possibility of synthesizing with nanoscale sizes. Additional properties of MOFs such as ultra-large surface-to-volume ratios, tunable pore size, high drug loading capacity, tunable structure and chemical composition, and potential for multiple postsynthetic modification make them ideal candidates for drug delivery. This review highlights recent research progress on MOF-based drug delivery systems (DDS), pointing out the evolution of these systems toward the development of theranostic nanoplatforms. Rather than a comprehensive review, representative recent examples are selected to illustrate such an evolution, and a critical discussion of the advantages and limitations of the different DDS types is given. Finally, the remaining challenges and future opportunities in this field are presented, highlighting that overcoming the current issues will pave the way toward the elusive dream of "personalized medicine." Graphical Abstract.


Simultaneous localization of two different tissue antigens based on the silver intensified PAP-DAB and on the traditional PAP-DAB method.
A new immunohistological method was elaborated for the light-microscopic detection of two tissue antigens in the same section. One of the antigens was visualized by the silver-gold intensified peroxidase-antiperoxidase complex (PAP)-diaminobenzidine (DAB) method resulting in a black coloured end-product. The other antigen was labelled by the brown reaction-product of the traditional PAP-DAB method. Both reaction-products are stable, electron-dense and they can clearly be distinguished from each other.


Intrasellar symptomatic salivary gland: case series and literature review.
Ectopic salivary glands have been found in both extracranial and intracranial locations, however, intrasellar symptomatic salivary gland is extremely rare and its clinical manifestation, radiological characteristics and outcome have not been systematically studied. Here we present a case series of intrasellar symptomatic salivary gland and perform a literature review to better characterize this disease. We retrospectively reviewed the data of three patients with intrasellar symptomatic salivary gland from our institutional and other cases available in literatures. Information for sex, age at diagnosis, clinical symptoms, radiological features, treatment strategy and prognosis were recorded. A total of 11 cases (including our own) were identified. There were three men and eight women, with an average age at diagnosis of 28.3 years. The peak incidence was in the second and the third decade (72.7% of all cases). The most common symptom was headache (81.8% of all patients). About 63.6% patients had one or more abnormal hormone levels, and prolactin was likely the most vulnerable hormone. The radiological appearances of intrasellar salivary gland were various, and four cases mimicked pituitary adenoma radiologically. All patients underwent transsphenoidal surgery with no mortality. Although intrasellar symptomatic salivary gland is rare, it should be considered in the differential diagnosis of intrasellar lesions. Preoperative diagnosis is challenging since it mimics pituitary neoplasm in clinical and radiological manifestations, and confirmation for this disease could only be conducted through pathological examination. Transsphenoidal surgical resection is the preferred therapy and the patient prognosis is usually good.


The role of pethidine in sedation for colonoscopy.
As sedation with a combination of benzodiazepines and opioids has been associated with an increased risk of serious cardiorespiratory events, it is rarely used in upper gastrointestinal endoscopy. The combination is, however, still commonly used in sedation for colonscopy and endoscopic retrograde cholangiopancreatography (ERCP). This study aimed to determine the benefits and disadvantages of adding pethidine to diazepam (Diazemuls) in the sedation for colonoscopic examinations. Seventy-one consecutive patients attending for routine colonoscopy were randomized in a double-blind, placebo-controlled manner to sedation with Diazemuls alone or Diazemuls with pethidine. The colonoscopist was at liberty to administrate any dose of Diazemuls considered appropriate for the patient's age and size, following which the patient was given an injection of pethidine 50 mg or a placebo. Both the patient and the endoscopist were blind to whether pethidine or a placebo was used. Adequacy of sedation was assessed and oxygen saturation recorded throughout the procedure. Oxygen was administered if the oxygen saturation fell below 92%. Patient satisfaction with sedation and recall of the procedure was recorded, by questionnaire, one day after examination. A total of 71 patients were entered into the trial. The two groups were similar with respect to age, sex, weight, alcohol intake, and state of health. An average of 18 mg of Diazemuls was given in the pethidine group and 19 mg in the placebo group. There was no statistically significant difference in the level of sedation obtained with the two regimens, although there was a tendency for the combination of Diazemuls with pethidine to give deeper sedation. Three patients were inadequately sedated with Diazemuls alone, requiring the code to be broken for pethidine to be given. Nine colonoscopies had to be abandoned due to patient intolerance in the group receiving Diazemuls, compared to five in the group receiving the combination regimen (p = 0.22). Twice as many patients receiving the combination of Diazemuls with pethidine required oxygen supplementation to maintain oxygen saturation during the procedure. This difference was highly statistically significant (p = 0.008). With the administration of supplemental oxygen, the average drop in oxygenation was similar in both groups. Satisfaction with sedation and recall for the procedure were similar in both groups. Although the patients had no preference for either regime, there may be an advantage in using the combination of Diazemuls and pethidine, as there was a trend for this combination to be preferred by the colonoscopists. The combination of a benzodiazepine with an opiate should be used with caution, however, as there was a greater requirement for oxygen in the group sedated by Diazemuls and pethidine.


Search for Alternative Control Strategies of Drosophila suzukii (Diptera: Drosophilidae): Laboratory Assays Using Volatile Natural Plant Compounds.
Drosophila suzukii (Diptera: Drosophilidae), is native to southeastern Asia and invaded Europe during the past decade. It causes serious economic damage in cherries and soft fruits. Control strategies rely on few insecticides with varying success. Due to environmental concern, the use of synthetic chemicals is restricted. Therefore, research effort is put into the quest for alternative substances applicable in chemical pest control. In laboratory assays, we tested 17 volatile plant compounds from different chemical classes for their contact toxicity, feeding modification, and oviposition repellency. Toxicity through contact with treated surfaces was evaluated after 1 h, 4 h, and 24 h; effects on food uptake were observed with capillary feeding (CAFE)-tests and oviposition trials compared egg numbers laid in raspberry medium with or without treated filter paper. Cinnamon oil and its components had the highest contact toxicity with an LC90 = 2-3%, whereas lemongrass oil, its main components, and farnesol were less toxic (LC90 = 7-9%), and geraniol was the least toxic. In CAFE tests, feeding stimulation was observed through 0.1% and 1% solutions of citronellol, lemongrass oil and farnesol. Cinnamon oil, cinnamaldhyde, and ethyl cinnamate were not consumed at a concentration of 1%. In the presence of citral, eugenol, and lemongrass oil, oviposition was reduced, and in the presence of limonene, no eggs were deposited. The natural products found most efficient in either bioassay will be further tested under field conditions.


Text-messaging to promote smoking cessation among individuals with opioid use disorder: quantitative and qualitative evaluation.
Individuals with opioid use disorder (OUD) who smoke cigarettes have high tobacco-related comorbidities, lack of access to tobacco treatment, lack of inclusion in smoking cessation trials, and remain understudied in the mobile health field. The purpose of this study was to understand patients' with OUD perceptions of 1) text message programs to promote smoking cessation, 2) content and features to include in such a program, and 3) how message content should be framed. From December 2018 to February 2019, we recruited 20 hospitalized individuals with a concurrent diagnosis of OUD and tobacco dependence at Boston Medical Center (BMC), the largest safety-net hospital in New England. We surveyed participants' cell phone use, their interest in a text message program to promote smoking cessation, and their reactions to and ratings of a series of 26 prototype texts. We then conducted open-ended interviews to elicit content and suggestions on how text message interventions can improve motivation to increase smoking cessation among individuals with OUD. The interviews also included open-ended inquiries exploring message ratings and message content, inquiries about preferences for message duration, frequency, and personalization. Quantitative analysis of questionnaire data indicated that the majority of participants owned a cell phone (95%, 19/20). Most participants (60%, 12/20) reported that they would be interested or very interested in receiving text messages about smoking cessation. Text messages about the health benefits of quitting were rated the highest among various categories of text messages. Qualitative analysis showed that almost every participant felt that text messages would help motivate smoking cessation given the support it would provide. This study demonstrates that individuals with OUD who smoke cigarettes perceive that a text message program designed to promote smoking cessation would motivate and support smoking cessation efforts. Our findings demonstrate that such a program is feasible as participants own cell phones, frequently send and receive text messages, and have unlimited text message plans. Findings from this study provide valuable insight into content and features to include when developing text message programs to address barriers to smoking cessation in individuals who have OUD and smoke cigarettes.


The measurement of hunger as a relative estimate of food available to populations of Nassarius pauperatus.
It is often difficult to estimate the amount of food available to an animal population, especially for species having unspecialised diets. The intertidal snail Nassarius pauperatus is omnivorous, feeding on both algae and animal carrion. Sampling the standing crops of carrion and algae was not thought to be a valid measure of the amount of food available to N. pauperatus. Instead, food availability was estimated indirectly, by measuring how hungry snails were.The percentage of snails feeding within 15 min of having been offered a meal of the bivalve Katelysia scalarina (Lamarck) was found to be a realiable and comparative estimate of food availability, and was measured in the field at 17 locations. The mean length of adult Nassarius pauperatus was negatively correlated with hunger, and at locations having high indices of hunger there was evidence of intraspecific competition for space to feed around items of carrion.These results are discussed and it is proposed that Nassarius pauperatus are surrounded by algal food of poor quality, with carrion being an important part of their diet for survival, growth and reproduction.


The harm of male-on-female rape: a response to David Benatar.
This article attacks David Benatar's claim made in his 2012 book, The Second Sexism: Discrimination Against Men and Boys, that when an individual is the victim of violence it does not matter whether his or her perpetrator is of the same or a different sex. By exploring two related yet distinct phenomena, well documented in the empirical psychological literature, that I call "shattering" and "fragmentation," I argue that when a woman is raped, it does matter that her rapist is male, given that her situation as a woman under patriarchy is partly constitutive of the harm that she suffers.


An Opportunity to END TB: Using the Sustainable Development Goals for Action on Socio-Economic Determinants of TB in High Burden Countries in WHO South-East Asia and the Western Pacific Regions.
The progress towards ending tuberculosis (TB) by 2035 is less than expected in 11 high TB burden countries in the World Health Organization South-East Asia and Western Pacific regions. Along with enhancing measures aimed at achieving universal access to quality-assured diagnosis, treatment and prevention services, massive efforts are needed to mitigate the prevalence of health-related risk factors, preferably through broader actions on the determinants of the "exposure-infection-disease-adverse outcome" spectrum. The aim of this manuscript is to describe the major socio-economic determinants of TB and to discuss how there are opportunities to address these determinants in an englobing manner under the United Nations Sustainable Development Goals (SDGs) framework. The national TB programs must identify stakeholders working on the other SDGs, develop mechanisms to collaborate with them and facilitate action on social-economic determinants in high TB burden geographical areas. Research (to determine the optimal mechanisms and impact of such collaborations) must be an integral part of this effort. We call upon stakeholders involved in achieving the SDGs and End TB targets to recognize that all goals are highly interlinked, and they need to combine and complement each other's efforts to end TB and the determinants behind this disease.


In the search for new anticancer drugs. 9. Synthesis and anticancer activity of spin-labeled analogues of N,N:N',N':N",N"-Tri-1,2-ethanediylphosphoric triamide and N,N:N',N':N",N"-tri-1,2-ethanediylphosphorothioic triamide.
A number of N,N:N',N':N",N"-tri-1,2- ethanediylphosphoric triamide (TEPA) and N,N:N',N':N",N"-tri-1,2- ethanediylphosphorothioic triamide (thio-TEPA) derivatives containing either two aziridine moieties (1a) or two (2-chloroethyl)amino functions (1b) and either a 2,2,6,6-tetramethylpiperidine, 1-oxy-2,2,6,6-tetramethylpiperidine or 1-hydroxy-2,2,6,6-tetramethylpiperidine component were synthesized and tested against lymphocytic leukemia P388 in mice. In a structure-activity comparison it was found that at optimum dose all compounds containing the nitroxyl radical were more active than the corresponding hydroxylamine derivatives. The open-chain compounds (1b) were less active than the corresponding aziridine ring compounds (1a). The replacement of the X = bridge in 1a with the X = N(CH3) group resulted in lowering of the anticancer activity.


Individual-level barriers to bariatric surgery from patient and provider perspectives: A qualitative study.
Less than 1% adults in the United States who meet body mass index criteria undergo bariatric surgery. Our objective was to identify patient and provider perceptions of individual-level barriers to undergoing bariatric surgery. Adults with severe obesity and obesity care providers described their experiences with the bariatric surgery care process in semi-structured interviews. Using conventional content analysis, individual-level barriers were identified within Andersen's Behavioral Model of Health Services Use. Of the 73 individuals interviewed, 36 (49%) were female, and 15 (21%) were non-white. Six individual-level barriers were identified: fear of surgery, fear of lifestyle change, perception that weight had not reached its "tipping point," concerns about dietary changes, lack of social support, and patient characteristics influencing referral. Patient and provider education should address patient fears of surgery and the belief that surgery is a "last resort." Bariatric surgery programs should strengthen social support networks for patients.


Adaptive switches in midbrain circuits.
In this issue of Neuron, Mysore and Knudsen (2012) describe a simple, anatomically supported circuit that can categorize stimuli into "strongest" and "others," regardless of their absolute strength. Such flexible categorization cannot be achieved by lateral inhibition alone but also requires that the inhibitory neurons reciprocally inhibit each other.


Effects of Hormone Therapy on Intraocular Pressure: The Women's Health Initiative-Sight Exam Study.
Previous studies suggest that hormone therapy favorably affects intraocular pressure (IOP). Here, we examined the association between hormone therapy use and IOP in the context of a large randomized trial. Secondary data analysis from a randomized controlled trial. We used data from the Women's Health Initiative-Sight Exam (WHISE). Women with prior hysterectomy received oral conjugated equine estrogen (0.625 mg/day) or placebo. Women with a uterus received estrogen plus progestin (medroxyprogesterone acetate 2.5 mg/day) or placebo. IOP was measured 5 years after randomization. Adjusted linear regression models were used to assess the association between hormone therapy and IOP. The WHISE included 1668 women in the estrogen-alone trial (aged 63-86, mean 72 years) and 2679 women in the estrogen-plus-progestin trial (aged 63-87, mean 72 years). In multivariate analyses, compared to placebo treatment, treatment with estrogen alone was associated with a 0.5 mm Hg reduction of the IOP in the right eye (95% CI: -0.8, -0.1, P = .005) and a 0.6 mm Hg (95% CI: -0.9, -0.3, P < .001) reduction of the IOP in the left eye. In the estrogen-plus-progestin trial, there was no significant difference in IOP between the treatment and placebo groups (P = .30 right eye and P = .43 left eye). This study represents an IOP analysis in the largest hormone trial available. Estrogen-alone therapy in postmenopausal women is associated with a small but significant IOP reduction of 0.5 mm Hg. The clinical significance of this small decrease remains to be determined.


Whole-genome analyses of loss of heterozygosity and methylation analysis of four tumor-suppressor genes in N-methyl-N'-nitro-N-nitrosoguanidine-induced rat stomach carcinomas.
N-Methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced rat stomach carcinomas are considered to be a good model for differentiated-type human stomach carcinomas. However, as for their molecular basis, only infrequent mutations of Catnb (beta-catenin) and Trp53 (p53) have been observed. Here, we carried out a whole-genome analysis of loss of heterozygosity (LOH) using 21 stomach carcinomas induced by MNNG in F(1) hybrids of ACI and BUF rats, and also analyzed promoter methylation of four tumor-suppressor genes. LOH analysis was performed using 130 polymorphic markers covering rat chromosomes 1-20 with an average interval of 20 Mbp. Despite adapting conditions so that LOH could be detected with up to a 50% contamination of stromal cells, no LOH was detected at any loci. CpG islands in putative promoter regions of four tumor-suppressor genes, Cdh1 (E-cadherin), Cdkn2a (p16), Mlh1, and Rassf1a, were analyzed by methylation-specific polymerase chain reaction (PCR). However, no methylation was detected. In contrast, the promoter region of Pgc (pepsinogen C), which lacks a CpG island, was methylated in all 21-cancer samples. These results indicated that LOH spanning a chromosomal region larger than 30-40 Mbp or silencing of Cdh1, Cdkn2a, Mlh1, and Rassf1a, was not involved in MNNG-induced rat stomach carcinomas. The search for other genes involved in these carcinomas needs to be continued.


Critical care management of the patient with acute spinal cord injury.
The critical care management of acute injury to the cervical spinal cord is discussed from the perspective of the pathophysiology of the injury process and its ramifications. Emphasis is placed upon resolution of cardiovascular derangements, spinal cord resuscitation, and respiratory support utilizing practical therapeutic interventions.


Extensor tendon triggering by impingement on the extensor retinaculum: a report of 5 cases.
We describe 4 patients who were treated for symptomatic triggering of the index and small fingers with pain on the dorsum of the hand and were found to have impingement of an extensor tendon on the extensor retinaculum at the wrist. All cases were treated by release of the extensor retinaculum and excision of pathologic structures.


Genetic characterization of Toxoplasma gondii strains by random amplified polymorphic DNA polymerase chain reaction.
The technique of Random Amplified Polymorphic DNA (RAPD) PCR has been used to detect DNA polymorphisms among Toxoplasma gondii strains. Seven arbitrary oligonucleotides (10-mer) were used as primers to amplify total genomic DNAs and significant genetic heterogeneity was detected among 11 T. gondii strains with different virulence for mice. The polymorphisms observed allowed relationship dendrograms of T. gondii strains to be constructed by PHYLIP and PAUP analyses. The genetic relationships of the T. gondii strains generated by 2 analyses using completely different assumptions were similar. Both analyses revealed 2 groups of T. gondii strains, one formed by the 6 virulent strains and the other formed by the 5 avirulent strains. This suggests that the genus Toxoplasma may actually contain 2 groups, correlated with their virulence, which have probably evolved independently following their initial separation. Significant polymorphisms were also detected between 2 different laboratory stocks of the T. gondii RH strain.


Effect of chronic treatment with angiotensin type 1 receptor antagonists on striatal dopamine levels in normal rats and in a rat model of Parkinson's disease treated with L-DOPA.
Beneficial effects of angiotensin type-1 receptor (AT1) inhibition have been observed in a number of brain processes mediated by oxidative stress and neuroinflammation, including Parkinson's disease. However, important counterregulatory interactions between dopamine and angiotensin systems have recently been demonstrated in several peripheral tissues, and it is possible that a decrease in dopamine levels due to AT1 inhibition may interfere with neuroprotective strategies. The present experiments involving rats with normal dopaminergic innervation indicate that chronic treatment with the AT1 antagonist candesartan does not significantly affect striatal levels of dopamine, serotonin or metabolites, as does not significantly affect motor behavior, as evaluated by the rotarod test. Interestingly, chronic administration of candesartan to normal rats induced a marked increase in dopamine D1 and a decrease in dopamine D2 receptor expression. In a rat model of Parkinson's disease treated with L-DOPA, no differences in striatal dopamine and serotonin levels were observed between candesartan-treated rats and untreated, which suggests that chronic treatment with candesartan does not significantly affect the process of L-DOPA decarboxylation and dopamine release in Parkinson's disease patients. Candesartan did not induce any differences in the striatal expression of dopamine D1 and D2 and serotonin 5-HT1B receptors in 6ydroxydopamine-lesioned rats treated with L-DOPA. The results suggest that chronic treatment with AT1 antagonists as a neuroprotective strategy does not significantly affect striatal dopamine release or motor behavior. This article is part of the Special Issue entitled 'The Synaptic Basis of Neurodegenerative Disorders'.


A Competing Risk-based Prognostic Model to Predict Cancer-specific Death of Patients with Spinal and Pelvic Chondrosarcoma.
Retrospective analysis. The aim of this study was to develop and validate a competing-risk-based prognostic model and a nomogram for predicting the three- and five-year probability of cancer-specific death (CSD) in patients with spinal and pelvic chondrosarcoma. The issue of competing risk has rarely been addressed and discussed in survival analysis of bone sarcoma. In addition, the Fine and Gray model, a more accurate method for survival analysis in the context of competing risk, has also been less reported in prognostic study of chondrosarcoma. A total of 623 patients with spinal or pelvic chondrosarcoma were identified from the SEER database and were divided into a training and a validation cohort. These two cohorts were used to develop and validate a prognostic model to predict the 3- and 5-year probability of CSD, considering non-CSD as competing risk. The C-index, calibration plot, and decision curve analysis were used to assess the predictive performance and clinical utility of the model. Older age (subdistribution hazards ratio [SHR]: 1.02, 95% confidence interval [CI]: 1.01∼1.03; P = 0.013), high grade (SHR: 2.68, 95% CI: 1.80∼3.99; P < 0.001), regional involvement (SHR: 1.66, 95% CI: 1.06∼2.58; P = 0.026), distant metastasis (SHR: 5.18, 95% CI: 3.11∼8.62; P < 0.001) and radical resection (SHR: 0.38, 95% CI: 0.24∼0.60; P < 0.001) were significantly associated with the incidence of CSD. These factors were used to build a competing-risk-based model and a nomogram to predict CSD. The C-index, calibration plot, and decision curve analysis indicated that the nomogram performs well in predicting CSD and is suitable for clinical use. A competing-risk based prognostic model is developed to predict the probability of CSD of patients with spinal and pelvic chondrosarcoma. This nomogram performs well and is suitable for clinical use.Level of Evidence: 4.


Resource partitioning of space and its relationship to body temperature in Anolis lizard populations.
An index is introduced that allows both the use and availability of space as a resource to be quantified in a manner that parallels the way that the use and availability of food resources are quantified in community ecology. This index provides the 'resource axis' for space resources that pertains to the thermal implications of micro-climate. The index is called the Grey Body Temperature Index (GBTI) and it is the equilibrium temperature that an inanimate reference object attains in the space being quantified. For this study the inanimate reference object is a grey lizard-shaped object weighing 5 grams. Formulae to calculate the GBTI from measurements of air temperature, wind speed and solar radiation are derived from an energy balance equation. The technique is illustrated with Anolis lizard populations from Grenada and St. Kitts. It is shown that the two lizard species in Grenada partition space as a resource with respect to GBTI and that the two species in St. Kitts do not. The determination of the availability of space along the GBTI axis is illustrated for a site in St. Maarten.


Correction to: Antithrombotic therapy after mitral valve repair: VKA or aspirin?
The original version of this article unfortunately contained a mistake in the author name. The co-author name should be Frederikus A. Klok instead of Frederik A. Klok. The original article has been corrected.


Predicting severe ischemic events after uncomplicated myocardial infarction by exercise testing and rest and exercise radionuclide ventriculography.
In 183 patients with uncomplicated myocardial infarction, exercise-induced angina, ST segment depression, decrease in ejection fraction, or inadequate increase in systolic blood pressure and low exercise tolerance were significantly associated with 4-year incidence of hard ischemic events. Only the onset of both ST segment depression and a decrease in left ventricular ejection fraction with exercise was an independent predictor. ST segment depression and decrease in left ventricular ejection fraction had low sensitivity (61% and 70%) and specificity (56% and 51%) for hard ischemic events, but specificity increased to 78% when both were present. During medical therapy, 22 of 53 patients with both ST segment depression and a decrease in left ventricular ejection fraction with exercise had an ischemic event (i.e., 48.1% 4-year probability on Kaplan-Meier analysis vs 19.2% in the remaining 130 patients [p < 0.0005]). Even if no single variable, derived from exercise testing, is a highly sensitive and specific predictor, specificity increases to a clinically relevant level by combining ST segment depression and a decrease in left ventricular ejection fraction with exercise, and in this way patients with recent infarction may be selected for coronary arteriography.


Quality control, and changing indications for lens implantation. The Seventh Binkhorst Medal Lecture-1982.
A total of 776 specimens yielded significant data on quality control and manufacturing defects in lens implants and on the biodegradation of lens implant and suture materials. Improved quality and an understanding of durability of materials in vivo has permitted wider safe use of intraocular lenses.


Identification of prognostic factors for the nonoperative treatment of stiff shoulder.
This study evaluated prognostic factors for the nonoperative treatment of stiff shoulder. Between June 2005 and May 2010, 497 stiff shoulders treated at our institute were included in this study. Multivariable analysis for recovery with Cox proportional hazard model was performed. The chief determining variable was pathogenesis (idiopathic, diabetic, post-traumatic) and confounding variables were age (49 or less, 50-59, 60 and above), sex, onset to visit interval (three months or less, four months or more), and external rotation (under 0°, 0° or more) or forward flexion (less than 90°, 90° or more) or internal rotation on the first visit. There were 356 idiopathic, 61 diabetic, and 80 post-traumatic stiff shoulders. Hazard ratio (HR) and 95 % confidence interval (CI) for recovery (lower HR means poor prognosis) was 0.54 (0.36-0.96) in the diabetic group (p = 0.007), and 0.92 (0.67-1.25) in the post-traumatic group (p = 0.58) compared with the idiopathic group. A positive correlation was observed in ages of 60 or over (HR 1.46, 95 % CI 0.86-1.65, p-value 0.02) and external rotation under 0° on the first visit (0.71, 0.53-0.96, 0.03). No correlations were observed in sex (p = 0.78) or onset to visit interval (p = 0.99). Similar results were obtained when forward flexion or internal rotation was used as a confounding variable. Diabetes mellitus and severely restricted joint motion on the first visit were poor prognostic factors and ages of 60 or over was a better prognostic factor.


Peroxidase activity in monocytes and tissue macrophages of mice.
A description is given of the distribution of peroxidatic (PO) activity in murine monocytes of blood and peritoneal cavity, and in murine macrophages residing in the unstimulated peritoneal cavity as well as in liver, spleen, bone marrow, and small intestine. In the monocytes, PO activity is restricted to some of the cytoplasmic granules; in the tissue (or resident) macrophages present in peritoneal cavity, liver, spleen, and small intestine, the PO activity is located in the nuclear envelope and the rough endoplasmic reticulum. Macrophages in the bone marrow are PO-negative. In the spleen and bone marrow, reticulum cells show PO activity in the nuclear envelope and the RER. Transitional forms between monocytes and tissue macrophages were not observed.


Sodium thiosulfate fails to increase the therapeutic index of intravenously administered cis-diamminedichloroplatinum (II) in mice bearing murine and human tumors.
Intravenous (i.v.) administration of sodium thiosulfate reduces the toxicity of cis-diamminedichloroplatinum (II) (CDDP). This effect, which allows the use of increased CDDP doses, has been exploited clinically in the intraperitoneal (i.p.) treatment of intraabdominal tumors. Recently, attempts have been made to treat extraperitoneal tumors by concurrent i.v. administration of CDDP and sodium thiosulfate. We have tested this approach in mice bearing systemic L1210 leukemia, s.c. growing Lewis lung carcinoma, C3H mammary carcinoma, and a human sarcoma growing in athymic nude mice. In all cases the antitumor activity of CDDP was substantially reduced in a manner dependent on the thiosulfate dose. Increased doses of CDDP, permitted by reduced toxicity in the presence of thiosulfate, raised the antitumor activity. However, the latter did not exceed that obtained by much lower doses in the absence of thiosulfate. The present experiments in animal models thus fail to support the clinical use of CDDP given i.v. together with its antidote, sodium thiosulfate.


"A disembodied voice over the telephone": a qualitative study of healthcare practitioners' experiences in geriatric medicine.
This study explored the experience of delivering care remotely among practitioners in a UK geriatric medicine clinic. Nine semi-structured interviews were conducted with consultants (n = 5), nurses (n = 2), a speech and language and an occupational therapist, and thematically analysed. Four themes developed; Challenges of remote consultations; Perceived advantages of remote consultations; Disruption of involvement of family members; Impact on care staff. Participants felt that rapport and trust had been more feasible to develop remotely than they had anticipated, although this was more challenging for new patients and those with cognitive or sensory impairments. While practitioners identified advantages of remote consultations, including involving relatives, saving time, and reducing anxiety, they also experienced disadvantages such as consultations feeling like a 'production line', missing visual cues and reduced privacy. Some participants felt their professional identity was threatened by the lack of face-to-face contact, linked to feeling that remote consultations are not suitable for frail older adults or those with cognitive deficits. Staff perceived barriers to remote consultations that went beyond practical concerns, and suggest support for building rapport, involving families, and protecting clinician identity and job satisfaction may be warranted.


Fate and distribution of sulfur-35 in yellow poplar and red maple trees.
Two deciduous tree species (yellow poplar and red maple) on Walker Branch Watershed (WBW), near Oak Ridge, Tennessee, were radiolabeled with 35S (87 day halflife) to study internal cycling, storage, and biogenic emission of sulfur (S). One tree of each species was girdled before radiolabeling to prevent phloem translocation to the roots, and the aboveground biomass was harvested prior to autumn leaf fall. Aboveground biomass, leaf fall, throughfall, and stemflow were sampled over a 13 to 24 week period. Sulfur-35 concentrations in tree leaves reached nearly asymptotic levels within 1 to 2 weeks after radiolabeling. Foliar leaching of 35S and leaf fall represented relatively unimportant return pathways to the forest soil. The final distribution of 35S in the nongirdled trees indicated little aboveground storage of S in biomass and appreciable (>60%) capacity to cycle S either to the belowground system by means of translocation or to the atmosphere by means of biogenic S emissions. Losses of volatile 35S were estimated from the amount of isotope missing (∼33%) in final inventories of the girdled trees. Estimated 35S emission rates from the girdled trees were ∼10-6 to ∼10-7 μCi cm-2 leaf d-1, and corresponded to an estimated gaseous S emission of approximately 0.1 to 1 μg S cm-2 leaf d-1. Translocation to roots was a significant sink for 35S in the red maple tree (40% of the injected amount). Research on forest biogeochemical S cycles should further explore biogenic S emissions from trees as a potential process of S flux from forest ecosystems.


Salvianolic acid A diminishes LDHA-driven aerobic glycolysis to restrain myofibroblasts activation and cardiac fibrosis via blocking Akt/GSK-3β/HIF-1α axis.
Myofibroblasts activation intensively contributes to cardiac fibrosis with undefined mechanism. Salvianolic acid A (SAA) is a phenolic component derived from Salvia miltiorrhiza with antifibrotic potency. This study aimed to interrogate the inhibitory effects and underlying mechanism of SAA on myofibroblasts activation and cardiac fibrosis. Antifibrotic effects of SAA were evaluated in mouse myocardial infarction (MI) model and in vitro myofibroblasts activation model. Metabolic regulatory effects and mechanism of SAA were determined using bioenergetic analysis and cross-validated by multiple metabolic inhibitors and siRNA or plasmid targeting Ldha. Finally, Akt/GSK-3β-related upstream regulatory mechanisms were investigated by immunoblot, q-PCR, and cross-validated by specific inhibitors. SAA inhibited cardiac fibroblasts-to-myofibroblasts transition, suppressed collage matrix proteins expression, and effectively attenuated MI-induced collagen deposition and cardiac fibrosis. SAA attenuated myofibroblasts activation and cardiac fibrosis by inhibiting LDHA-driven abnormal aerobic glycolysis. Mechanistically, SAA inhibited Akt/GSK-3β axis and downregulated HIF-1α expression by promoting its degradation via a noncanonical route, and therefore restrained HIF-1α-triggered Ldha gene expression. SAA is an effective component for treating cardiac fibrosis by diminishing LDHA-driven glycolysis during myofibroblasts activation. Targeting metabolism of myofibroblasts might occupy a potential therapeutic strategy for cardiac fibrosis.


[Fertilizer Effect on Yield and Quality of Gentiana crassicaulis].
To find the best fertilizer formula of yield-quality-fertilizer effect, and to establish a new comprehensive evaluation-fertilizer effect method. Grey related degree analysis and DTOPSIS method were combined to comprehensive evaluate the 11 indexes of 14 fertilizer formulas of nitrogen, phosphorus and potassium, in order to explore the best dosage and ratio. The fertilizer requirements of the factors which contribute to the yield and quality of Gentiana crassicaulis were different and the most appropriate rates of nitrogen, phosphorus and potassium fertilizer were 21.49~42.03 kg/667 m2, 49.53~65.75 kg/667 m2 and 26.34~ 42.97 kg/667 m2, respectively. The fertilizer formula in this study can be widely used in the standardized plantation of Gentiana crassicaulis.


Debate: short-term orthodontics.
Debate based on an opinion article published in the British Dental Journal entitled 'Truth or consequences: the potential implications of short-term cosmetic orthodontics for general dental practitioners' by R. A. C. Chate ( 2013; 215: 551-553; www.nature.com/bdj/journal/v215/n11/full/sj.bdj.2013.1140.html).


Patterns of salt transport and factors affecting typical shrub in desert-oases transition areas.
Soil salinization and water deficits are considered the primary factors limiting economic development and environmental improvement in arid areas. However, there remains limited knowledge of the adaptability of typical shrubs to salinization of desert areas in arid zones. This study was conducted in a desert oasis transition zone (Tarim River, China), aiming to investigate: i) the spatial-temporal changes in soil salinity; ii) the interactions between the pedoenvironment vs typical shrub (Calligonum mongolicum). The van Genuchten soil salinity retention ensemble model (TVGSSREM-3D) was developed to simulate variations in soil water-salt transport in the desert-oasis zone and to accurately explain the main factors influencing Calligonum mongolicum desert-oases transition areas. The results showed that monthly average salinity ranged from 2.0 to 8.0 g kg-1, with a peak in August (9.17 g kg-1). The presence of human activities (Salt Drainage Canal) and the distribution of Calligonum mongolicum resulted in a clear spatial salinity zonation. Moreover, analysis of environmental indicators using the TVGSSREM-3D model revealed strong correlations between the distribution of salinity in Calligonum mongolicum desert-oases transition areas and groundwater depth (GD), minimum relative humidity (MRH), and water vapor pressure (WVP). These findings provide a scientific basis for stabilizing, restoring, and reconstructing the ecosystem of the oasis-desert transition zone.


Inhibition by protein kinase-C inhibitor and cycloheximide of phorbol ester- and epidermal growth factor-induced arachidonic acid metabolism in cultured porcine thyroid cells.
In an attempt to elucidate possible mechanism(s) for stimulated arachidonic acid metabolism by phorbol 12-myristate 13-acetate (PMA) and epidermal growth factor (EGF) in porcine thyroid cells, we examined the effects of protein kinase inhibitors, isoquinolinesulfonamide derivatives (H-7 and HA-1004), and cycloheximide. The production of PGE2 stimulated by either PMA or EGF was strongly inhibited by H-7, with an ID50 value of approximately 20 to 25 mumol/L in each case, as well as by cycloheximide, with an ID50 value of less than 0.5 micrograms/mL in each case. In contrast, 100 mumol/L of HA-1004 showed less inhibition of PGE2 production provocated by either PMA or EGF. On the other hand, PGE2 production in basal or stimulated condition by exogenously added arachidonic acid, was inhibited to an even lesser extent by both H-7 and cycloheximide. The EGF- and PMA-stimulated release of 3H-arachidonic acid from the cells was also strongly inhibited by H-7 and cycloheximide. These results suggest an induction of synthesis of some proteins responsible for the release of arachidonic acid, which might be attributed to protein kinase-C activation in arachidonic acid metabolism stimulated by PMA or EGF. Moreover, PGE2 production was potently induced by PMA and slightly by EGF in the cyclooxygenase-inactivated cells by acetyl salicylate pretreatment, which also suggests that both agents might induce the synthesis of cyclooxygenase in cultured porcine thyroid cells, although we did not measure its activity.


Developmental epidemiology of depressive disorders.
Definitions, understanding, and treatment of childhood depressive disorders are changing. The last 40 years have seen a move from questioning whether depression even existed in younger children to evidence-based descriptive models. The field is now moving toward developmentally informed multifactorial models that more accurately reflect the complexity, heterogeneity, and dimensionality of depressive disorders. Knowledge about genetic, temperamental, and developmental risks has increased. Inability to self-regulate seems to be common in depressive and related disorders. Positive modulation can be promoted through experiences, psychotherapies, and, possibly, medications. The authors provide an overview of childhood depressive disorders with emphasis on the developmental/etiologic underpinnings.


Screening for Colorectal Cancer With Fecal Immunochemical Testing With and Without Postpolypectomy Surveillance Colonoscopy: A Cost-Effectiveness Analysis.
Population-based screening to prevent colorectal cancer (CRC) death is effective, but the effectiveness of postpolypectomy surveillance is unclear. To evaluate the additional benefit in terms of cost-effectiveness of colonoscopy surveillance in a screening setting. Microsimulation using the ASCCA (Adenoma and Serrated pathway to Colorectal CAncer) model. Dutch CRC screening program and published literature. Asymptomatic persons aged 55 to 75 years without a prior CRC diagnosis. Lifetime. Health care payer. Fecal immunochemical test (FIT) screening with colonoscopy surveillance performed according to the Dutch guideline was simulated. The comparator was no screening or surveillance. FIT screening without colonoscopy surveillance and the effect of extending surveillance intervals were also evaluated. CRC burden, colonoscopy demand, life-years, and costs. FIT screening without surveillance reduced CRC mortality by 50.4% compared with no screening or surveillance. Adding surveillance to FIT screening reduced mortality by an additional 1.7% to 52.1% but increased lifetime colonoscopy demand by 62% (from 335 to 543 colonoscopies per 1000 persons) at an additional cost of €68 000, for an increase of 0.9 life-year. Extending the surveillance intervals to 5 years reduced CRC mortality by 51.8% and increased colonoscopy demand by 42.7% compared with FIT screening without surveillance. In an incremental analysis, incremental cost-effectiveness ratios (ICERs) for screening plus surveillance exceeded the Dutch willingness-to-pay threshold of €36 602 per life-year gained. When using a parameter set representing low colorectal lesion prevalence or when colonoscopy costs were halved or colorectal lesion incidence was doubled, screening plus surveillance became cost-effective compared with screening without surveillance. Limited data on FIT performance and background CRC risk in the surveillance population. Adding surveillance to FIT screening is not cost-effective based on the Dutch ICER threshold and substantially increases colonoscopy demand. Extending surveillance intervals to 5 years would decrease colonoscopy demand without substantial loss of effectiveness. Alpe d'HuZes, Dutch Cancer Society, and Stand Up To Cancer.


Cerebrospinal fluid camk2a levels at baseline predict long-term progression in multiple sclerosis.
Multiple sclerosis (MS) remains a highly unpredictable disease. Many hope that fluid biomarkers may contribute to better stratification of disease, aiding the personalisation of treatment decisions, ultimately improving patient outcomes. The objective of this study was to evaluate the predictive value of CSF brain-specific proteins from early in the disease course of MS on long term clinical outcomes. In this study, 34 MS patients had their CSF collected and stored within 5 years of disease onset and were then followed clinically for at least 15 years. CSF concentrations of 64 brain-specific proteins were analyzed in the 34 patient CSF, as well as 19 age and sex-matched controls, using a targeted liquid-chromatography tandem mass spectrometry approach. We identified six CSF brain-specific proteins that significantly differentiated MS from controls (p < 0.05) and nine proteins that could predict disease course over the next decade. CAMK2A emerged as a biomarker candidate that could discriminate between MS and controls and could predict long-term disease progression. Targeted approaches to identify and quantify biomarkers associated with MS in the CSF may inform on long term MS outcomes. CAMK2A may be one of several candidates, warranting further exploration.


Defective karyogamy in meiotic segregants of a Candida utilis-Saccharomyces cerevisiae hybrid.
Meiotic segregants derived from a Candida utilis--Saccharomyces cerevisiae hybrid obtained by protoplast fusion were crossed to several standard S. cerevisiae laboratory strains. Random spore and tetrad analysis suggested that in these segregants, karyogamy is impaired and internuclear chromosome transfer occurs.


Serum microRNAs as potential biomarkers of mandibular prognathism.
The aim of the study was to determine whether the expression levels of specific circulating serum microRNAs (miRNAs) are associated with mandibular prognathism (MP). Sixty subjects in the early permanent dentition stage and 23 in the mixed dentition stage with MP were identified. Sixty-eight normal control subjects in the early permanent dentition stage and 24 in the mixed dentition stage were recruited for comparison. According to the microarray-based expression profiling, four serum miRNAs (let-7i-3p, miR-595, miR-16-2-3p, and miR-367-5p) were validated. In the MP groups, let-7i-3p was significantly over-expressed in subjects in the early permanent (P < 0.0005) and mixed (P < 0.001) dentitions, and miR-595 was significantly under-expressed (P < 0.004) in subjects in the early permanent (P < 0.004) and mixed (P < 0.0005) dentitions, compared with normal control groups. Multiple logistic regression analysis and receiver operating characteristic curve analysis revealed that let-7i-3p and miR-595 were able to significantly discriminate MP subjects from normal controls. Let-7i-3p and miR-595 could be potential, non-invasive biomarkers for the accurate early detection and diagnosis of MP, which may result in improved clinical management.


Economic evaluation of laboratory diagnostic test types in Covid-19 epidemic: A systematic review.
Corona 2 virus (SARS-CoV-2) is known as the causative agent of COVID-19 disease; the World Health Organization (WHO) declared it an epidemic on March 11, 2020. The Joint Guidelines of the Centers for Disease Control and Prevention (CDC) and the WHO including social distancing, the use of face masks, emphasis on hand washing, quarantine, and using diagnosis tests have been used widely, but the value of diagnostic interventions to prevent the transmission of SARS-CoV-2 is unclear. We compared the economic evaluation of different laboratory diagnostic interventions with each other and also with implementing the conservative CDC & WHO guidelines. Electronic searches were conducted on PubMed, Embase, Science Direct, Scopus, Cochrane Library, Web of Knowledge, NHSEED, NHS Health Technology assessment (CRD), and Cost-Effectiveness Analysis Registry databases. Related articles were reviewed from January 2020 to the end of November 2021. Out of 1791 initial studies, 13 articles had the inclusion criteria. According to the Consolidated Health Economic Evaluation Reporting Standards (CHEERS) checklist, ten studies were of excellent quality, and the remaining two studies were of very good quality. Most studies were cost-effectiveness analysis studies. The entered studies had different time horizons. Diagnostic tests reviewed in the studies included real-time polymerase chain reaction (RT-PCR) test, immunoglobulin G (IgG) & Antigen, point of care tests. Although polymerase chain reaction (PCR) testing improves the quality of life and survival for patients with infected Covid-19 based on its greater effectiveness compared to standard protection protocols, due to the high cost of this intervention, it has been considered a cost-effective method in some countries. Since most studies have been conducted in developed countries, it unquestionably does not make sense to extend these results to low-income and developing countries. Therefore further studies are required in low-income and developing countries to evaluate the cost-effectiveness of laboratory-based diagnostic methods (RT-PCR) of covid-19 in variable prevalence of infectious cases.


Effects of capsaicin on inflammation and on the substance P content of nervous tissues in rats with adjuvant arthritis.
Capsaicin (20-80 mg/kg, s.c.) reduced the inflammatory response to inoculation with Mycobacterium butyricum in the rat. The effect was apparent within 24 h, was partial, persisted for well over 20 days, and occurred irrespective of whether capsaicin was administered before or after the onset of inflammation, or at the time when the pathology reached peak. Capsaicin also attenuated the increase in substance P content in sciatic nerve, saphenous nerve, dorsal root ganglia, dorsal roots, and dorsal spinal cord (L4, L5) which occurs in rats with adjuvant arthritis. The data are consistent with a possible role of substance P in the peripheral manifestations of adjuvant arthritis.


Left Atrial Size May Predict Blood Pressure Improvement After Ganglionated Plexi Ablation in Addition to Pulmonary Vein Isolation.
Hypertension (HTN) and atrial fibrillation (AF) commonly co-exist. An improvement in control of HTN in a subset of patients undergoing AF ablation was previously demonstrated by our group. In the present study, we aimed to assess whether left atrial (LA) size based on transthoracic echocardiography may predict the patients who demonstratebetter HTN improvement after ganglionated plexus ablation (GPA) in addition to pulmonary vein isolation (PVI). This was a retrospective chart review of patients with AF and HTN who underwent GPA+PVI. Patients were divided into 2 groupsbased on LA size: Patients with normal LA size and patients with LA enlargement. Systolic blood pressure (SBP) levelswere compared at baseline, and 3, 6, and 12 months post-ablation. The primary endpoints of the study weremean systolic blood pressure change compared between groups from baseline to 12-months, as well as the absolute difference in systolic blood pressure at 12 months follow-up.Medical therapy for HTN was also assessed before the procedure, and at 12 months post-procedure. 53 patients (37 with LA enlargement, 16 with normal LA size) met inclusion criteria. At 12 months follow-up, SBP was 136.46 ± 22.38 mmHg in patients with LA enlargementand 118.25 ± 9.81 mmHg in patients with normal LA size (estimated difference of 19.04 ± 6.98 mmHg, p = 0.01). Patients with normal LA size were on significantly fewer anti-hypertensive agents at 12 months (2.33 ± 1.49 vs. 1.44 ± 1.21, p < 0.05). In patients undergoing PVI+GP ablation, normal LA size may predict HTN improvement at 12 months post-procedure. Normal LA size may identify hypertensive AF patients for whom autonomic modulation could be an effective therapy.


Prevalence of HBV, HDV and HCV hepatitis markers in HIV-positive patients.
Since HIV infection could condition the natural history of parenterally transmitted viral hepatitis (HBV, HCV, HDV), with possibly differing effects in different risk groups, we decided to retrospectively examine sera from a cohort of 637 HIV seropositive patients in different stages of infection, seen from 1985 to 1992, to study the prevalence and temporal course of these infections. Virological markers of HBV, HCV and HDV were determined by ELISA and RIBA methods. The severity of HIV infection was higher in homosexuals than in drug addicts. Prevalence of antiHBc antibodies was 82% in drug addicts and 77% in homosexuals, whereas antiHCV antibodies prevalence was 72% in drug addicts and only 7% in homosexuals (p < 0.000001). When only antiHBc-positive patients were considered, there was a significant difference in antiHBs antibodies between drug addicts (DA) and homosexuals (OR for DA 0.29, 95% CI 0.08/0.83, p = 0.02), suggesting that drug addicts are less able to produce a protective response. This fact cannot be explained by the severity of HIV infection (which was higher among the homosexual group) and suggests some immunodepressive effect of drug abuse. Delta infection was only detected in the drug addict group, and the prevalence was low. Finally, we cannot confirm the interference of HCV infection with the speed of HBsAg clearance: in this study the prevalence of HBsAg was almost the same in HCV-positive and negative patients.


The membrane-embedded segment of cytochrome b5 as studied by cross-linking with photoactivatable phospholipids. II. The nontransferable form.
Rabbit liver cytochrome b5 was reconstituted into vesicles containing dipalmitoyl-sn-glycero-3-phosphocholine and the radioactively labeled photoactivatable 1-palmitoyl-2-omega-(m-diazirinophenoxy)undecanoyl-sn-glycero-3-phosphocholine (PC-I) by the cholate-gel filtration method. The cytochrome inserted predominantly in the nontransferable form (Preparation I). In addition, reconstituted vesicles containing asymmetrically distributed radioactively labeled PC-I were prepared by the use of the phosphatidylcholine transfer protein. This preparation (Preparation II) contained [14C]PC-I in the outer leaflet and [3H]PC-I predominantly in the inner leaflet. Following photolysis of the labeled vesicle preparations, the purified cross-linked products were analyzed for the distribution of the sites of cross-links. In Preparation I, a broad distribution (Asn-105 to Met-130) was found, similar to that observed with the transferable form. On the other hand, the peptide fragments obtained from the asymmetrically labeled preparation II showed 14C/3H ratios which decreased in going from Asn-105 to Met-130. We conclude that the membrane-embedded segment in the nontransferable form of cytochrome b5 spans the bilayer.


Blood vessel transplantation to bone.
In dogs an artery, a vein, and a vascular bundle were transplanted into intact bone, isolated bone segments, necrotized bone, and homografts of bone. Active proliferation of new blood vessels and formation of new bone occurred in all instances where the vascular bundle was used, when the vein was implanted into the intact bone and isolated bone, and only when the artery was implanted into intact bone. As a result of these experiments, vascular bundle transplantation has been performed in patients with Kienboeck's disease, and avascular necrosis of the scaphoid, the femoral head, the talus, and other conditions.


Surgical treatment of stages IB and IIA invasive carcinoma of the cervix by radical abdominal hysterectomy.
From 1963 to 1977, 349 radical abdominal hysterectomies with bilateral pelvic lymphadenectomy were performed for Stage IB (331 patients) and Stage IIA (18 patients) cervical cancer at the New York Medical College, with no operative deaths. Definitive diagnosis was obtained from the biopsy specimen in 281 patients. Twenty-nine patients were pregnant when the diagnosis was established. The average operating time was 4 hours and 48 minutes with an average blood loss of 900 ml. Eleven fistulas were noted: ureterovaginal, 7; vesicovaginal, 3; rectovaginal, 1. Since 1972, there have been no fistulas in 130 radical hysterectomies. Metastatic carcinoma of the regional lymph nodes was discovered in 27 patients for an incidence of 7.7%. Postoperative total pelvic external irradiation was utilized in 40 patients (27 with positive nodes, 10 with microscopic carcinoma in vascular channels, and an additional three patients with an inadequate vaginal extirpation margin). A total of 62% of those patients with poor prognostic criteria receiving postoperative irradiation are alive and well. Two hundred nineteen patients have been followed up for at least 5 years and the survival rate was 90%. Our data support the view that radical abdominal hysterectomy with bilateral pelvic lymphadenectomy is the treatment of choice for patients with Stages IB and IIA cervical cancer in the nonpregnant state, unless there are major medical contraindications.


Noninvasive monitoring of infection and rejection after lung transplantation.
The survival rate following lung transplantation is among the lowest of all solid-organ transplants, and current diagnostic tests often fail to distinguish between infection and rejection, the two primary posttransplant clinical complications. We describe a diagnostic assay that simultaneously monitors for rejection and infection in lung transplant recipients by sequencing of cell-free DNA (cfDNA) in plasma. We determined that the levels of donor-derived cfDNA directly correlate with the results of invasive tests of rejection (area under the curve 0.9). We also analyzed the nonhuman cfDNA as a hypothesis-free approach to test for infections. Cytomegalovirus is most frequently assayed clinically, and the levels of CMV-derived sequences in cfDNA are consistent with clinical results. We furthermore show that hypothesis-free monitoring for pathogens using cfDNA reveals undiagnosed cases of infection, and that certain infectious pathogens such as human herpesvirus (HHV) 6, HHV-7, and adenovirus, which are not often tested clinically, occur with high frequency in this cohort.