Upregulated miR-17 Regulates Hypoxia-Mediated Human Pulmonary Artery Smooth Muscle Cell Proliferation and Apoptosis by Targeting Mitofusin 2.
BACKGROUND Pulmonary arterial hypertension (PAH) is a fatal disease characterized by impaired regulation of pulmonary artery vascular growth and remodeling. Aberrant expression of miR-17 has been shown to be involved in the pathogenesis of PAH, but its underlying molecular mechanism has not been elucidated. MATERIAL AND METHODS Mitofusin 2 (MFN2) expression was determined by qRT-PCR. The protein expression levels of MFN2, proliferating cell nuclear antigen (PCNA), and pro-apoptotic protein cleaved Caspase-3 were measured using Western blot analysis. Cell proliferation and apoptosis were assessed by CellTiter-Glo reagent and flow cytometry, respectively. Caspase-3/7 activity was measured using an Apo-ONE Homogeneous Caspase-3/7 assay kit. The regulation of miR-17 on MFN2 expression was assessed using luciferase reporter assay system. RESULTS miR-17 expression was upregulated in human pulmonary artery smooth muscle cells (hPASMCs) treated with hypoxia and lung tissues of PAH patients. Inhibition of miR-17 suppressed hypoxia-induced proliferation and promoted apoptosis in hPASMCs. miR-17 inhibited MFN2 expression by binding to its 3'-UTR. Decreased cell viability and increased apoptosis and Caspase-3 activity were observed in the anti-miR-17 + siNC group compared with the anti-miR-NC + siNC group. The expression of cleaved Caspase-3 was upregulated and the expression of PCNA was downregulated in the anti-miR-17 + siNC group. Moreover, these alterations were attenuated by knockdown of MFN2. CONCLUSIONS miR-17 regulates proliferation and apoptosis in hPASMCs through MFN2 modulation. We found that miR-17 acts as a potential regulator of proliferation and apoptosis of hPASMCs, and that it might be developed as a promising new strategy for the treatment of PAH.


Chlorogenic acid-chitosan copolymers: Synthesis, characterization and application in O/W emulsions for enhanced β-carotene stability.
In this study, chlorogenic acid-chitosan (CA-CS) copolymers were prepared with varying Chitosan (CS): chlorogenic acid (CA)ratios and characterized for their water solubility, antioxidant capacity, and emulsions stability. Results showed that CA-CS samples exhibited up to 90.5 % increase in DPPH scavenging efficiency and 20 % increase in hydroxyl radical scavenging efficiency compared to CS alone. CA-CS copolymers used to stabilize oil in water (O/W) emulsions, which were evaluated for their potential in encapsulating and protecting β-carotene. Microscopic observations revealed homogeneous spherical droplets in stable emulsions, suggesting effective interfacial structures. The selected CA-CS-stabilized O/W emulsions demonstrated encapsulation efficiencies of 74.8 % and 75.26 % for β-carotene. The CA-CS stabilized O/W emulsions provided the most effective protection against β-carotene degradation under UV exposure, retaining over 80 % of β-carotene content after 12 h of testing. These findings indicate that CA-CS-based O/W emulsions show promise as carriers and protectors for bioactive compounds, due to their improved antioxidant capacity, emulsions stability, and protection against degradation.


The Pathogenesis Based on the Glymphatic System, Diagnosis, and Treatment of Idiopathic Normal Pressure Hydrocephalus.
Idiopathic normal pressure hydrocephalus (iNPH) is a rare neurological disorder with no clear prevalence factors and is a significant danger to the elderly. The intracranial glymphatic system is the internal environment that maintains brain survival and metabolism, and thus fluid exchange changes in the glymphatic system under various pathological conditions can provide important insights into the pathogenesis and differential diagnosis of many neurodegenerative diseases such as iNPH. iNPH can be diagnosed using a combination of clinical symptoms, imaging findings and history, and cerebrospinal fluid biomarkers due to the glymphatic system disorder. However, only few researchers have linked the two. Shunt surgery can improve the glymphatic system disorders in iNPH patients, and the surgical approach is determined using a combination of clinical diagnosis and trials. Therefore, we have composed this review to provide a future opportunity for elucidating the pathogenesis of iNPH based on the glymphatic system, and link the glymphatic system to the diagnosis and treatment of iNPH. The review will provide new insights into the medical research of iNPH.


5-Bromo-8-methoxy-1-methyl-beta-carboline, an alkaloid from the New Zealand marine bryozoan Pterocella vesiculosa.
A new alkaloid, 5-bromo-8-methoxy-1-methyl-beta-carboline (1), has been isolated from the New Zealand marine bryozoan Pterocella Vesiculosa. Structural elucidation was achieved through NMR spectroscopic and mass spectrometric analysis, and a single-crystal X-ray diffraction analysis of 1 was performed. The biological activity of 1 was assessed against P388 murine leukemia cells and three microorganisms and compared with that of a number of related beta-carboline alkaloids.


Lumbar disc degeneration in relation to occupation.
The aim was to study risk factors of lumbar disc degeneration demonstrable with magnetic resonance imaging (MRI) with special emphasis on occupational load and back accidents. The subjects in this cross-sectional study were 53 machine drivers, 51 construction carpenters, and 60 municipal office workers aged 40-45 years. Data on possible risk factors were available from current structured questionnaires and for 4 and 7 years in retrospect. The prevalence of lumbar disc degeneration L2/L3-L5/S1 was determined with MRI. An increased risk was found for posterior disc bulges among the carpenters and for anterior disc bulges among the machine drivers, but decreased signal intensity was not related to occupation. Car driving was also associated with anterior disc bulges. All signs of disc degeneration were related to a history of back accidents. Disc degeneration was not related to body height, overweight, smoking, or the frequency of physical exercise. Occupational load affects the risk of disc degeneration of the lumbar spine. Accidental back injuries and motor vehicle driving are associated with an increased risk of disc degeneration. Anterior and posterior disc bulges seem to be related to different types of physical loads.


Protocol of Co-Culture of Human Osteoblasts and Osteoclasts to Test Biomaterials for Bone Tissue Engineering.
New biomaterials and scaffolds for bone tissue engineering (BTE) applications require to be tested in a bone microenvironment reliable model. On this assumption, the in vitro laboratory protocols with bone cells represent worthy experimental systems improving our knowledge about bone homeostasis, reducing the costs of experimentation. To this day, several models of the bone microenvironment are reported in the literature, but few delineate a protocol for testing new biomaterials using bone cells. Herein we propose a clear protocol to set up an indirect co-culture system of human-derived osteoblasts and osteoclast precursors, providing well-defined criteria such as the cell seeding density, cell:cell ratio, the culture medium, and the proofs of differentiation. The material to be tested may be easily introduced in the system and the cell response analyzed. The physical separation of osteoblasts and osteoclasts allows distinguishing the effects of the material onto the two cell types and to evaluate the correlation between material and cell behavior, cell morphology, and adhesion. The whole protocol requires about 4 to 6 weeks with an intermediate level of expertise. The system is an in vitro model of the bone remodeling system useful in testing innovative materials for bone regeneration, and potentially exploitable in different application fields. The use of human primary cells represents a close replica of the bone cell cooperation in vivo and may be employed as a feasible system to test materials and scaffolds for bone substitution and regeneration.


[Effect of early initiation of continuous renal replacement therapy based on the KDIGO classification on the prognosis of critically ill patients with acute kidney injury].
To investigate the impact of early initiation of continuous renal replacement therapy (CRRT) based on "Kidney Disease: Improving Global Outcomes (KDIGO)" classification on the prognosis of critically ill patients with acute kidney injury (AKI). A retrospective analysis of clinical data of patients diagnosed as AKI in Department of Critical Care Medicine of Zhejiang Provincial People's Hospital from January 2011 to January 2015 was conducted. All patients included should be 18 years old or older, having stayed in intensive care unit (ICU) for more than 48 hours, and received CRRT. All subjects were divided into three groups according to their renal function before CRRT according to the KDIGO-AKI guideline: AKI-stage 1 group, AKI-stage 2 group and AKI-stage 3 group. The general condition, original disease, severity of disease, duration of mechanical ventilation, the length of ICU or hospital stay, 28-day survival rate and in-hospital mortality rate were compared among these three groups. Additionally, risk factors for the 28-day survival rate and hospital mortality of critically ill patients with AKI were screened by logistic regression analysis. A total of 258 critically ill patients with AKI were enrolled, with 64 cases in AKI-stage 1 group, 62 cases in AKI-stage 2 group, and 132 cases in AKI-stage 3 group. 116 patients survived with 28-day survival rate of 44.96%. 154 patients died with hospital mortality 59.69%. The precipitating factors of AKI in all three groups (stage 1, stage 2, and stage 3) were similar, with sepsis, heart failure and poisoning (drugs or poison) being the main triggers for AKI, accounting for 35.66%, 19.38% and 13.18%, respectively. There were significant differences in the rate of vasoactive agent usage (31.25%, 41.94%, 50.00%, χ2 = 6.241, P = 0.044), acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score (20.87±7.37, 17.19±7.02, 22.58±7.95, F = 5.292, P = 0.006) and sequential organ failure assessment (SOFA) score (8.41±3.46, 6.22±2.43, 9.58±3.71, F = 10.328, P = 0.000), while there was no significant difference in gender, age, primary disease, time from ICU admission to the beginning of CRRT, mean arterial pressure (MAP), lactate level or 24-hour lactate clearance rate (LCR), mechanical ventilation time, the length of ICU or hospital stay, 28-day survival rate or hospital mortality among these three groups (all P > 0.05). According to the logistic regression analysis, time from ICU admission to start of CRRT and lactate level were the independent risk factors for 28-day survival rate or hospital mortality of critically ill patients with AKI [odds ratio (OR) for 28-day survival rate was 0.850 and 0.774, 95% confidence interval (95%CI) was 0.752-0.960 and 0.638-0.940, P value was 0.009 and 0.010, respectively; OR for hospital mortality was 0.884 and 0.756, 95%CI was 0.781-1.000 and 0.610-0.939, P value was 0.049 and 0.011, respectively]. Early initiation of CRRT based on KDIGO-AKI classification could not improve the prognosis of critically ill patients with AKI, the optimal timing of RRT for such patients remains to be further explored.


Comparing staffing levels in the Online Survey Certification and Reporting (OSCAR) system with the Medicaid Cost Report data: are differences systematic?
This study had two goals: (a) to assess the validity of the Online Survey Certification and Reporting (OSCAR) staffing data by comparing them to staffing measures from audited Medicaid Cost Reports and (b) to identify systematic differences between facilities that over-report or underreport staffing in the OSCAR. We merged the 2002 Texas Nursing Facility Cost Report, the OSCAR for Texas facilities surveyed in 2002, and the 2003 Area Resource File. We eliminated outliers in the OSCAR using three decision rules, resulting in a final sample size of 941 of the total of 1,017 non-hospital-based facilities. We compared OSCAR and Medicaid Cost Report staffing measures for three staff types. We examined differences between facilities that over-reported or underreported staffing levels in the OSCAR by using logistic regression. Average staffing levels were higher in the OSCAR than in the Medicaid Cost Report data. The two sets of measures exhibited correlations ranging between 0.5 and 0.6. For-profit and larger facilities consistently over-reported registered nurse staffing levels. Factors associated with increased odds of over-reporting licensed vocational nursing or certified nursing assistant staffing were lower Medicare or Medicaid censuses and less market competition. Facility characteristics associated with over-reporting were consistent across different levels of over-reporting. Underreporting was much less prevalent. Certain types of facilities consistently over-report staffing levels. These reporting errors will affect the validity of consumer information systems, regulatory activities, and health services research results, particularly research using OSCAR data to examine the relationship between staffing and quality. Results call for a more accurate reporting system.


The Impact of a Computerized Clinical Decision Support Tool on Inappropriate Clostridium difficile Testing.
OBJECTIVE To evaluate the effectiveness of a computerized clinical decision support intervention aimed at reducing inappropriate Clostridium difficile testing DESIGN Retrospective cohort study SETTING University of Pennsylvania Health System, comprised of 3 large tertiary-care hospitals PATIENTS All adult patients admitted over a 2-year period INTERVENTION Providers were required to use an order set integrated into a commercial electronic health record to order C. difficile toxin testing. The order set identified patients who had received laxatives within the previous 36 hours and displayed a message asking providers to consider stopping laxatives and reassessing in 24 hours prior to ordering C. difficile testing. Providers had the option to continue or discontinue laxatives and to proceed with or forgo testing. The primary endpoint was the change in inappropriate C. difficile testing, as measured by the number of patients who had C. difficile testing ordered while receiving laxatives. RESULTS Compared to the 1-year baseline period, the intervention resulted in a decrease in the proportion of inappropriate C. difficile testing (29.6% vs 27.3%; P=.02). The intervention was associated with an increase in the number of patients who had laxatives discontinued and did not undergo C. difficile testing (5.8% vs 46.4%; P<.01) and who had their laxatives discontinued and underwent testing (5.4% vs 35.2%; P<.01). We observed a nonsignificant increase in the proportion of patients with C. difficile related complications (5.0% vs 8.9%; P=.11). CONCLUSIONS A C. difficile order set was successful in decreasing inappropriate C. difficile testing and improving the timely discontinuation of laxatives. Infect Control Hosp Epidemiol 2017;38:1204-1208.


High arctic heath soil respiration and biogeochemical dynamics during summer and autumn freeze-in - effects of long-term enhanced water and nutrient supply.
In High Arctic NE Greenland, temperature and precipitation are predicted to increase during this century, however, relatively little information is available on the role of increased water supply on soil CO 2 efflux in dry, high arctic ecosystems. We measured soil respiration (Rsoil ) in summer and autumn of 2009 in combination with microbial biomass and nutrient availability during autumn freeze-in at a dry, open heath in Zackenberg, NE Greenland. This tundra site has been subject to fully factorial manipulation consisting of increased soil water supply for 14 years, and occasional nitrogen (N) addition in pulses. Summer watering enhanced Rsoil during summer, but decreased Rsoil in the following autumn. We speculate that this is due to intensified depletion of recently fixed plant carbon by soil organisms. Hence, autumn soil microbial activity seems tightly linked to growing season plant production through plant-associated carbon pools. Nitrogen addition alone consistently increased Rsoil , but when water and nitrogen were added in combination, autumn Rsoil declined similarly to when water was added alone. Despite several freeze-thaw events, the microbial biomass carbon (C) remained constant until finally being reduced by ~60% in late September. In spite of significantly reduced microbial biomass C and phosphorus (P), microbial N did not change. This suggests N released from dead microbes was quickly assimilated by surviving microbes. We observed no change in soil organic matter content after 14 years of environmental manipulations, suggesting high ecosystem resistance to environmental changes.


A unified model for the coevolution of resistance, tolerance, and virulence.
We present a general host-parasite model that unifies previous theory by investigating the coevolution of virulence, resistance, and tolerance, with respect to multiple physiological, epidemiological, and environmental parameters. Four sets of new predictions emerge. First, compared to virulence coevolving with resistance or tolerance, three-trait coevolution promotes more virulence and less tolerance, and broadens conditions under which pure defenses evolve. Second, the cost and efficiency of virulence and the epidemiological rates are the key factors of virulence coevolving with resistance and tolerance. Maximum virulence evolves for intermediate infection rate, at which coevolved levels of resistance and tolerance are both high. The influence of host and parasite background mortalities is strong on the evolution of defenses and weak on the coevolution of virulence. Third, evolutionary correlations between defenses can switch sign along single-parameter gradients. The evolutionary trade-off between resistance and tolerance may coevolve with virulence that either increases or decreases monotonically, depending on the underlying parameter gradient. Fourth, despite global attractiveness and stability of coevolutionary equilibria, not-so-rare and not-so-small mutations can beget large variation in virulence and defenses around equilibrium, in the form of transient "evolutionary spikes." Implications for evolutionary management of infections are discussed and directions for future research are outlined.


Obstructive sleep apnea is associated with preserved bone mineral density in healthy elderly subjects.
Chronic intermittent hypoxia (IH) acts as a stimulator of mesenchymal stem cell (MSC) mobilization, intensifying osteoblast formation in animal models. The recurrence of apnea and oxygen desaturation in obstructive sleep apnea (OSA) may mimic experimental models of IH. We hypothesized that in elderly with OSA, apnea-related IH may mobilize MSCs and thereby prevent the age-related decline in osteogenesis. This study explored the relationship between OSA and bone mineral density (BMD), and the effect of IH on BMD, in a large sample of elderly subjects. There were 833 volunteers age 68.6 ± 0.8 y (59% women). Each participant underwent evaluation of BMD at lumbar spine and femoral sites by dual-energy x-ray absorptiometry (DEXA) as well as clinical and polygraphic examinations. OSA was diagnosed on the basis of an apnea-hypopnea index (AHI) ≥ 15. There were 55% of the participants who presented with OSA, and these subjects were predominantly male and overweight. Compared with subjects without OSA, those with OSA had a higher femoral and spinal BMD (P < 0.001). Body mass index (BMI), AHI, and oxygen desaturation index (ODI) (P < 0.01) were significantly related to BMD. After adjustment for sex, BMI, metabolic values, and hypertension, multiple regression analysis showed a significant association between femoral and lumbar T scores and both daily energy expenditure (P < 0.001) and ODI (P = 0.007). In elderly subjects, the presence of obstructive sleep apnea is associated with higher bone mineral density, with oxygen desaturation index being a significant determinant of bone metabolism. These results suggest that apnea-related intermittent hypoxia may stimulate the bone remodeling process in older population. NCT 00759304 and NCT 00766584.


Structural and functional analysis of Mre11-3.
The Mre11, Rad50 and Nbs1 proteins make up the conserved multi-functional Mre11 (MRN) complex involved in multiple, critical DNA metabolic processes including double-strand break repair and telomere maintenance. The Mre11 protein is a nuclease with broad substrate recognition, but MRN-dependent processes requiring the nuclease activity are not clearly defined. Here, we report the functional and structural characterization of a nuclease-deficient Mre11 protein termed mre11-3. Importantly, the hmre11-3 protein has wild-type ability to bind DNA, Rad50 and Nbs1; however, nuclease activity was completely abrogated. When expressed in cell lines from patients with ataxia telangiectasia-like disorder (ATLD), hmre11-3 restored the formation of ionizing radiation-induced foci. Consistent with the biochemical results, the 2.3 A crystal structure of mre11-3 from Pyrococcus furiosus revealed an active site structure with a wild-type-like metal-binding environment. The structural analysis of the H85L mutation provides a detailed molecular basis for the ability of mre11-3 to bind but not hydrolyze DNA. Together, these results establish that the mre11-3 protein provides an excellent system for dissecting nuclease-dependent and independent functions of the Mre11 complex.


Goal-Driven Cognition and Functional Behavior: The Fundamental-Motives Framework.
Fundamental motives have direct implications for evolutionary fitness and orchestrate attention, memory, and social inference in functionally specific ways. Motivational states linked to self-protection and mating offer illustrative examples. When self-protective motives are aroused, people show enhanced attention to, and memory for, angry male strangers; they also perceive out-group members as especially dangerous. In contrast, when mating motives are aroused, men show enhanced attention to and memory for attractive members of the opposite sex; mating motives also lead men (but not women) to perceive sexual arousal in attractive members of the opposite sex. There are further functionally specific consequences for social behavior. For example, self-protective motives increase conformity among both men and women, whereas mating motives lead men (but not women) to engage in anticonformist behavior. Other motivational systems trigger different adaptive patterns of cognitive and behavioral responses. This body of research illustrates the highly specific consequences of fitness-relevant motivational states for cognition and behavior, and highlights the value of studying human motivation and cognition within an evolutionary framework.


Confusion assessment method: a systematic review and meta-analysis of diagnostic accuracy.
Delirium is common in the early stages of hospitalization for a variety of acute and chronic diseases. To evaluate the diagnostic accuracy of two delirium screening tools, the Confusion Assessment Method (CAM) and the Confusion Assessment Method for the Intensive Care Unit (CAM-ICU). We searched MEDLINE, EMBASE, and PsychInfo for relevant articles published in English up to March 2013. We compared two screening tools to Diagnostic and Statistical Manual of Mental Disorders IV criteria. Two reviewers independently assessed studies to determine their eligibility, validity, and quality. Sensitivity and specificity were calculated using a bivariate model. Twenty-two studies (n = 2,442 patients) met the inclusion criteria. All studies demonstrated that these two scales can be administered within ten minutes, by trained clinical or research staff. The pooled sensitivities and specificity for CAM were 82% (95% confidence interval [CI]: 69%-91%) and 99% (95% CI: 87%-100%), and 81% (95% CI: 57%-93%) and 98% (95% CI: 86%-100%) for CAM-ICU, respectively. Both CAM and CAM-ICU are validated instruments for the diagnosis of delirium in a variety of medical settings. However, CAM and CAM-ICU both present higher specificity than sensitivity. Therefore, the use of these tools should not replace clinical judgment.


Phase Ib Study of Atezolizumab Plus Interferon-α with or without Bevacizumab in Patients with Metastatic Renal Cell Carcinoma and Other Solid Tumors.
This Phase Ib study combined programmed death-ligand 1 inhibitor, atezolizumab, with other immunomodulatory agents in locally advanced and metastatic solid tumors. Arms B-D evaluated atezolizumab plus interferon-α, with/without vascular endothelial growth factor inhibitor, bevacizumab, in renal cell carcinoma (RCC) and other solid tumors. Arm B predominantly recruited patients with previously treated RCC or melanoma to receive atezolizumab plus interferon α-2b. Arm C investigated atezolizumab plus polyethylene glycol (PEG)-interferon α-2a in previously treated RCC. Arm D evaluated atezolizumab plus PEG-interferon α-2a and bevacizumab. Primary objectives were safety and tolerability; secondary objectives included clinical activity. Combination therapy was well tolerated, with safety profiles consistent with known risks of individual agents. The most frequent treatment-related toxicities were fatigue, chills, and pyrexia. The objective response rate (ORR) in arm B was 20.0% overall and 17.8% in patients with previously treated checkpoint inhibitor-naive RCC (n = 45). No responses were reported in arm C. The highest ORR in arm D was 46.7% in patients with treatment-naive RCC (n = 15). Data showed preliminary clinical activity and acceptable tolerability of atezolizumab plus interferon α-2b in patients with previously treated checkpoint inhibitor-naive RCC and of atezolizumab plus PEG-interferon α-2a and bevacizumab in patients with treatment-naive RCC.


Identification of the Ebola virus glycoprotein as the main viral determinant of vascular cell cytotoxicity and injury.
Here we defined the main viral determinant of Ebola virus pathogenicity; synthesis of the virion glycoprotein (GP) of Ebola virus Zaire induced cytotoxic effects in human endothelial cells in vitro and in vivo. This effect mapped to a serine-threonine-rich, mucin-like domain of this type I transmembrane glycoprotein, one of seven gene products of the virus. Gene transfer of GP into explanted human or porcine blood vessels caused massive endothelial cell loss within 48 hours that led to a substantial increase in vascular permeability. Deletion of the mucin-like region of GP abolished these effects without affecting protein expression or function. GP derived from the Reston strain of virus, which causes disease in nonhuman primates but not in man, did not disrupt the vasculature of human blood vessels. In contrast, the Zaire GP induced endothelial cell disruption and cytotoxicity in both nonhuman primate and human blood vessels, and the mucin domain was required for this effect. These findings indicate that GP, through its mucin domain, is the viral determinant of Ebola pathogenicity and likely contributes to hemorrhage during infection.


Counts of MSC in the Bone Marrow of Young and Old CBA Mice after a Single Exposure to Osteogenic Stimuli (Curettage, BMP-2 Injection) or Antigens (S. typhimurium Antigenic Complex) and in Heterotopic Bone Marrow Transplants.
The efficiency of cloning and the content of multipotent stromal cells (MSC) in the femoral bone marrow of intact CBA mice was 1.5 times less in old mice (24-36 months) than in young ones (2-3 months). The concentration of osteogenic MSC was higher in old vs. young mice (42±3 vs. 22±2%, respectively). Changes in the total counts of MSC and concentrations of osteogenic MSC in response to osteogenic (curettage, BMP-2) and immunogenic stimuli (S. typhimurium antigenic complex) were similar in young and old mice in comparison with intact controls of respective age. The counts of the total pool of bone marrow MSC and pool of osteogenic MSC in response to osteogenic stimuli were 1.5-2 times less in old vs. young mice. This difference seemed to be a result of age-specific decrease of their bone marrow count but not of age-specific decrease of the MSC functional activity, this leading to a decrease in the transplantability of bone marrow stromal tissue of old mice. Comparison of transplantations "old donor - young recipient" vs. "young donor - young recipient" demonstrated a decrease in the count of nuclear cells (1.8 times), size of bone capsule (2-fold), efficiency of MSC cloning (1.6 times), count of MSC per transplant (2.9 times), and count of osteogenic MSC per transplant (3.3 times). The concentrations of osteogenic MSC in transplants from young and old donors leveled in young recipients, that is, seemed to be regulated by the host. Serum concentrations of IL-10 and TNF-α in intact old mice were at least 2.9 and 2 times higher than in young animals, while the concentrations of almost all the rest studied cytokines (IL-2, IL-5, GM-CSF, IFN-γ, IL-4, IL-12) were lower. Presumably, the decrease in the content of bone marrow MSC and in transplantability of bone marrow stromal tissue in old mice were caused by exhaustion of the MSC pool as a result of age-specific chronic inflammation. These data indicated a close relationship between age-specific changes in the stromal tissue and immune system.


Zhangfei is a potent and specific inhibitor of the host cell factor-binding transcription factor Luman.
Host cell factor (HCF) was initially discovered as a cellular co-factor required for the activation of herpes simplex virus immediate early gene expression by the virion associated transactivator VP16. HCF also participates in a variety of cellular processes, although the mechanism of its action is not known. VP16 binds to HCF through a 4-amino acid motif (EHAY), which closely resembles the HCF binding domain of two cellular basic leucine-zipper proteins, Luman and Zhangfei. Luman is a powerful transcription factor that, in transient expression assays, activates promoters containing cAMP or unfolded protein response elements (UPRE). In contrast, Zhangfei neither binds consensus recognition elements for basic leucine-zipper proteins nor does it activate promoters containing them. Here we show that Zhangfei suppresses the ability of Luman to activate transcription. HCF appeared to be required for efficient suppression. A mutant of Zhangfei, which was unable to bind HCF, was impaired in its ability to suppress Luman. Zhangfei did not suppress ATF6, a transcription factor closely related to Luman but that does not bind HCF, unless the HCF binding motif of Luman was grafted onto it. Zhangfei inhibited the HCF-dependent activation of a UPRE-containing promoter by a Gal4-Luman fusion protein but was unable to inhibit the HCF-independent activation by Gal4-Luman of a promoter that contained Gal4 binding motifs. Binding of HCF by Zhangfei was required for the co-localization of Luman and Zhangfei to nuclear domains, suggesting that HCF might target the proteins to a common location.


Anti-GAD Antibody-associated Syndrome Presenting with Limb Myoclonus.
The clinical spectrum of anti-glutamic acid decarboxylase (GAD) antibody-associated neurologic syndromes is expanding, with focal, generalized, and atypical forms. We describe a 59-year-old female showing continuous right lower limb myoclonus and mild encephalopathy. These symptoms started 2 weeks prior to evaluation. The patient had great improvement with intravenous steroids. An autoantibody panel was positive for anti-GAD. Various clinical manifestations, including myoclonus, may relate to anti-GAD antibodies. The treatment options available include symptomatic drugs, intravenous immunoglobulin, steroids, and other immunosuppressant agents.


Comparison of prognostic values of high-sensitivity cardiac troponin T and N-terminal prohormone brain natriuretic peptide to assess mortality in elderly inpatients.
Studies have shown that increases in low-level high-sensitivity cardiac troponin T (hs-cTnT) and N-terminal prohormone brain natriuretic peptide (NT-proBNP) in the elderly population lead to high risk of adverse clinical outcomes, such as mortality. In this study, associations between the two biomarkers and long-term mortality in elderly patients hospitalized for medical conditions other than acute cardiovascular events were investigated. A comparison of the predictive value of hs-cTnT and NT-proBNP for all-cause mortality was conducted. A cohort of 715 elderly inpatients free of acute cardiovascular events was initially recruited. Based on tertiles of baseline hs-cTnT and NT-proBNP levels, the elderly were arranged into low, middle, and high groups. The mortality of each group was observed and the predictive values of hs-cTnT and NT-proBNP compared. Additionally, all-cause mortality was analyzed for hs-cTnT and NT-proBNP combined. There were 135 (18.9%) all-cause mortality cases identified during follow-up (median 47 months). The upper tertile of hs-cTnT was significantly associated with an increased risk of mortality (HR 3.29, 95% CI 1.85-5.85), even after adjustment for potential confounders. However, there were no significant differences observed in mortality rates among the three NT-proBNP groups after adjustment for potential confounders (HR 1.77, 95% CI 0.83-3.17). Compared to NT-proBNP, hs-cTnT was a better predictor of mortality, as area under curves for hs-cTnT and NT-proBNP at 60 months were 0.712 (95% CI 0.616-0.809) and 0.585 (95% CI 0.483-0.6871), respectively. Also, the combination of the two cardiac biomarkers did not render a better predictive value for mortality than hs-cTnT alone. Unlike hs-cTnT, baseline NT-proBNP levels failed to show an independent association with all-cause mortality in hospitalized elderly without acute cardiovascular events. hs-cTnT provided significant prognostic value for mortality in the study cohort.


Repair of a perforated sinus membrane with a subepithelial palatal conjunctive flap: technique report and evaluation.
The maxillary sinus grafting procedure has proven to be an acceptable modality for bone augmentation to provide a base for endosseous implants, routinely used for the rehabilitation of posterior maxilla. Perforation of the membrane is the most common complication in this type of procedure. This paper presents a technique for repairing a perforated Schneiderian membrane with a conjunctive connective tissue graft harvested from the palate and shows the histological and radiographic evaluation of the results. Ten consecutives cases with the occurrence of membrane perforation were included in this study. All were repaired with a flap of tissue removed from the palatine portion near to the surgical site. The technique is demonstrated through a clinical case. The results showed successful integration of 88.8% of the implants after 12 months from prosthesis installation. Histological evaluation of the samples showed that the use of nanocrystalized hydroxyapatite showed an adequate stimulation of boné neoformation within 6 months. Radiographic evaluation revealed a small apical implant bone loss, not compromising their anchorages and proservation. Thus, it can be concluded that the use of conjunctive technique with collected palate flap for sealing the perforation of the membrane of the sinus may have predictable result.


T-Cell Activation Independently Associates With Immune Senescence in HIV-Infected Recipients of Long-term Antiretroviral Treatment.
Aging-associated noncommunicable comorbidities are more prevalent among human immunodeficiency virus type 1 (HIV)-infected individuals than among HIV-uninfected individuals. Residual HIV-related chronic immune activation and senescence may increase the risk of developing comorbidities. Immune phenotyping, thymic output, and telomere length were assessed in 94 HIV-infected individuals who were aged >45 years and receiving antiretroviral therapy (ART; cases) and 95 age-matched uninfected controls. Cases had lower CD4(+) T-cell counts, higher CD8(+) T-cell counts, and increased levels of immune activation (ie, increased soluble CD14 [sCD14] level and increased percentages of CD38(+)HLA-DR(+) cells among both CD4(+) and CD8(+) T cells), regulatory T cells, and percentage of programmed cell death 1 (PD-1)-expressing cells among CD4(+) T cells. Immune senescence levels (ie, percentages of CD27(-)CD28(-) cells or CD57(+) cells) were comparable between cases and controls. Peripheral blood mononuclear cells from cases had shorter telomeres but increased single-joint T-cell receptor excision circle content and CD31(+) naive CD4(+) T cells. Although cytomegalovirus (CMV) antibody titers were higher in cases, CMV-specific T-cell responses were comparable between cases and controls. T-cell senescence in cases was independently associated with T-cell activation but not with CMV-specific immune responses. Despite long-term receipt of ART, HIV-infected adults had higher levels of immune activation, regulatory T cells, and PD-1-expressing CD4(+) cells and shorter telomeres. The increased soluble CD14 levels and percentage of CD38(+)HLA-DR(+) cells among CD4(+) T cells correlated with shorter telomeres and increased regulatory T-cell levels. This suggests that HIV influences immune function irreversibly, with several pathways that are persistently abnormal during effective ART. Therapies aimed at improving immune health during ART are needed.


Noncovalent interactions in the gas-phase conformers of anionic iduronate (methyl 2-o-sulfo-α-L-iduronate): variation of subconformer versus ring conformer energetics for a prototypical anionic monosaccharide studied using computational methods.
The conformational preferences of the prototypical anionic monosaccharide (methyl 2-O-sulfo-α-L-iduronate) have been studied at various computational levels to investigate the energetic variation of 17 subconformers associated with the (4)C(1), (2)S(0), (5)S(1), and (1)C(4) ring conformers. These calculations include the first fully optimized MP2 calculations that have been performed for an anionic sugar system, and therefore allow an assessment of the performance of a group of DFT functionals (B3LYP, PW91PW91, and M05-2X) for treating the noncovalent dispersion and anomeric effects that are present in this system. We find that the recently developed M05-2X functional of Truhlar and co-workers [Y. Zhao, N. E. Schultz, D. G. Truhlar, J. Chem. Theory Comput., 2006, 2, 364] reproduces the MP2 results most closely, thus indicating that it may well be suitable for computational studies of larger ionic saccharides. Most importantly, the results presented indicate that it is crucial to consider the subconformers (which correspond to rearrangements of the sugar-ring side-chains) of the main ring-conformers in order to obtain a reliable overview of the potential energy surface of such systems. We find that the lowest isolated (gas-phase) conformer corresponds to a (4)C(1) chair conformer, which displays a pair of strong C(3)-OH···SO(3)(-) and OMe···SO(3)(-) electrostatic hydrogen-bonding interactions, coupled with a looser C(4)-OH···SO(3)(-) interaction. Overall, the relative energies of the subconformers appear to be straightforwardly related to the number of hydrogen-bonding interactions that each conformer displays among its pendant functional groups.


Understanding chronic pain complicating disability: finding meaning through focus group methodology.
Although extensive literature exists on the experiences of people living with chronic nonmalignant pain as a primary condition, little is known about the phenomenon of pain as it is experienced by the person with a chronic disabling condition. This focus group study explored the experience of disability-related pain among 32 people with multiple sclerosis (MS) living in the community. Thematic analysis of transcripts revealed 4 broad conceptualizations of the experience of living with chronic MS-related pain. The first theme, pain is pervasive, described the overwhelming and intrusive presence of pain in daily life. Participants related the physical, emotional, and social consequences of living with chronic, disability-related pain, which caused them great loss and restriction. The second theme resonated around feelings that nobody understands. The participants sought understanding and validation of their pain experiences by family, friends, and healthcare professionals and struggled with difficult issues concerning the legitimacy and invisibility of their pain. The third theme, I'm fine, referred to the propensity of participants to keep pain private from others. In part because of its contested nature, participants at times concealed their pain from others to avoid conflict and maintain some semblance of their former lives. The final theme that emerged was always a factor in the equation. The participants shared how MS had transformed their worlds into ones in which pain and discomfort had become a normal part of everyday life, requiring careful negotiation and planning to undertake activities and prevent exacerbation. It is vital that healthcare providers give people with MS opportunities to talk about pain and pain-related concerns, validate their experiences, and provide interventions that enable self-management. Clinicians are encouraged to challenge their own meanings and expectations about disability-related pain so that therapeutic interventions can be facilitated.