Phosphoinositide signaling in human brain.
The phosphoinositide signal transduction system constitutes one of the primary means for intercellular communication in the central nervous system, but only recently has this system been studied in human brain. Although some investigations have studied phosphoinositide signaling in slices from biopsied human brain, due to the limited access to such material a greater number of studies have utilized membranes prepared from postmortem human brain. With membranes exposed to exogenous labeled phosphoinositides, activation of phospholipase C with calcium, with G-proteins stimulated by GTP gamma S or NaF, or with several receptor agonists, have demonstrated that all of the components of the phosphoinositide system are retained in human brain membranes and are responsive to appropriate stimuli. Investigators have begun to examine the effects of neurological (Alzheimer's disease, epilepsy, Parkinson's disease) and psychiatric (schizophrenia, major depression, bipolar affective disorder) diseases on the activity of the phosphoinositide system. Alzheimer's disease has been studied to the greatest extent and a severe deficit in phosphoinositide signaling has been identified in most studies. In addition, brain regionally selective deficits in G-protein function associated with phosphoinositide signaling have been reported in subjects with major depression or with bipolar affective disorder, and in the latter an ameliorative effect of the therapeutic drug lithium was identified. Although significant progress has been achieved in studying the phosphoinositide system in human brain, many issues remaining to be addressed are discussed in this review. With carefully controlled studies, it appears that much will be learned in the near future about the phosphoinositide signal transduction system in human brain and the effects of a variety of disorders on its function.


Pulmonary arterial hypertension: updates in epidemiology and evaluation of patients.
Group 1 pulmonary hypertension (or pulmonary arterial hypertension) is a rare, highly complex, and progressive disorder that is incurable and ultimately can lead to premature death. PAH causes significant physical, social, work, and emotional burdens among affected patients and their caregivers. Early diagnosis and initiation of treatment is required for best outcomes; however, the clinical presentation of PAH is nonspecific and frequently overlaps with several other conditions, often leading to a delay in diagnosis or misdiagnosis. In the past decades, increased understanding of the pathobiology of PAH has led to changes in its definition. Additionally, contemporary PAH registries have shown greater survival rates among patients with PAH and have allowed for the development of risk calculator tools that are now used to drive therapeutic goals. To date, multiple PAH-specific therapies have been developed, and all currently target one of 3 pathways that contribute to the endothelial dysfunction pathogenesis of PAH (prostacyclin, endothelin, and nitric oxide pathways). Because PAH is classified into 7 subgroups, it is essential that individuals are grouped appropriately for the efficacy of treatment and avoidance of harm. As health-related quality of life for PAH is multifactorial, it is important that patients are involved in the clinical decision-making process and have access to multidisciplinary care. The purpose of this review is to update healthcare professionals on the management of PAH with the most current information on epidemiology, pathophysiology, clinical presentation, and diagnostic considerations.


Rho GDP dissociation inhibitor-mediated disruption of Rho GTPase activity impairs lens fiber cell migration, elongation and survival.
To explore the role of the Rho GTPases in lens morphogenesis, we overexpressed bovine Rho GDP dissociation inhibitor (Rho GDI alpha), which serves as a negative regulator of Rho, Rac and Cdc42 GTPase activity, in a lens-specific manner in transgenic mice. This was achieved using a chimeric promoter of delta-crystallin enhancer and alpha A-crystallin, which is active at embryonic day 12. Several individual transgenic (Tg) lines were obtained, and exhibited ocular specific phenotype comprised of microphthalmic eyes with lens opacity. The overexpression of bovine Rho GDI alpha disrupted membrane translocation of Rho, Rac and Cdc42 GTPases in Tg lenses. Transgenic lenses also revealed abnormalities in the migration pattern, elongation and organization of lens fibers. These changes appeared to be associated with impaired organization of the actin cytoskeleton and cell-cell adhesions. At E14.5, the size of the Rho GDI alpha Tg lenses was larger compared to wild type (WT) and the central lens epithelium and differentiating fibers exhibited an abnormal increase of bromo-deoxy-uridine incorporation. Postnatal Tg eyes, however, were much smaller in size compared to WT eyes, revealing increased apoptosis in the disrupted lens fibers. Taken together, these data demonstrate a critical role for Rho GTPase-dependent signaling pathways in processes underlying morphogenesis, fiber cell migration, elongation and survival in the developing lens.


Cornification of keratinocytes is associated with differential changes in the catalytic activity and the immunoreactivity of transglutaminase-1.
Transglutaminase 1 (TGM1) plays an essential role in skin barrier formation by cross-linking proteins in differentiated keratinocytes. Here, we established a protocol for the antibody-dependent detection of TGM1 protein and the parallel detection of TGM activity. TGM1 immunoreactivity initially increased and co-localized with membrane-associated TGM activity during keratinocyte differentiation. TGM activity persisted upon further differentiation of keratinocytes, whereas TGM1 immunoreactivity was lost under standard assay conditions. Pretreatment of tissue sections with the proteases trypsin or proteinase K enabled immunodetection of TGM1 in cornified keratinocytes, indicating that removal of other proteins was a prerequisite for TGM1 immunolabeling after cornification. The increase of TGM activity and subsequent loss of TGM1 immunoreactivity could be replicated in HEK293T cells transfected with TGM1, suggesting that protein cross-linking mediated by TGM1 itself may lead to reduced recognition of TGM1 by antibodies. To screen for proteins potentially regulating TGM1, we performed Virotrap experiments and identified the CAPNS1 subunit of calpain as an interaction partner of TGM1. Treatment of keratinocytes and TGM1-transfected HEK293T cells with chemical inhibitors of calpain suppressed transglutamination. Our findings suggest that calpain contributes to the control of TGM1-mediated transglutamination and proteins cross-linked by transglutamination mask epitopes of TGM1.


Putative roles of soluble trophic factors in facial nerve regeneration, target reinnervation, and recovery of vibrissal whisking.
It is well-known that, after nerve transection and surgical repair, misdirected regrowth of regenerating motor axons may occur in three ways. The first way is that the axons enter into endoneurial tubes that they did not previously occupy, regenerate through incorrect fascicles and reinnervate muscles that they did not formerly supply. Consequently the activation of these muscles results in inappropriate movements. The second way is that, in contrast with the precise target-directed pathfinding by elongating motor nerves during embryonic development, several axons rather than a single axon grow out from each transected nerve fiber. The third way of misdirection occurs by the intramuscular terminal branching (sprouting) of each regenerating axon to culminate in some polyinnervation of neuromuscular junctions, i.e. reinnervation of junctions by more than a single axon. Presently, "fascicular" or "topographic specificity" cannot be achieved and hence target-directed nerve regeneration is, as yet, unattainable. Nonetheless, motor and sensory reinnervation of appropriate endoneurial tubes does occur and can be promoted by brief nerve electrical stimulation. This review considers the expression of neurotrophic factors in the neuromuscular system and how this expression can promote functional recovery, with emphasis on the whisking of vibrissae on the rat face in relationship to the expression of the factors. Evidence is reviewed for a role of neurotrophic factors as short-range diffusible sprouting stimuli in promoting complete functional recovery of vibrissal whisking in blind Sprague Dawley (SD)/RCS rats but not in SD rats with normal vision, after facial nerve transection and surgical repair. Briefly, a complicated time course of growth factor expression in the nerves and denervated muscles include (1) an early increase in FGF2 and IGF2, (2) reduced NGF between 2 and 14days after nerve transection and surgical repair, (3) a late rise in BDNF and (4) reduced IGF1 protein in the denervated muscles at 28days. These findings suggest that recovery of motor function after peripheral nerve injury is due, at least in part, to a complex regulation of nerve injury-associated neurotrophic factors and cytokines at the neuromuscular junctions of denervated muscles. In particular, the increase of FGF2 and concomittant decrease of NGF during the first week after facial nerve-nerve anastomosis in SD/RCS blind rats may prevent intramuscular axon sprouting and, in turn, reduce poly-innervation of the neuromuscular junction.


[Impaired superoxide dismutase activity and decreased content of lipid peroxides in hepatocellular carcinoma tissue].
Superoxide dismutase (SOD) activity and content of lipid peroxides (LPO) in human hepatocellular carcinoma (HCC) tissue were studied. It was observed that SOD activity and LPO content in HCC tissue were lower than those in normal liver tissue (P less than 0.001 respectively). The contents of copper, zinc and manganese in HCC tissue were also lower than those in normal liver tissue (P less than 0.001, P less than 0.05). Furthermore, LPO content in necrotic HCC tissue was higher than that in non-necrotic HCC tissue (P less than 0.05). The results suggest that deficiency of copper, zinc and manganese in HCC tissue may be a contributing factor that leads to impairment of SOD activity. Decreased SOD activity in liver cancer cells was a negative feedback of the multiplication of cancer cells loss of lipid peroxidation explains the malignancy of HCC, and enhanced lipid peroxidation in liver cancer cells may cause the necrosis of cancer cells.


Evaluating severity of white matter lesions from computed tomography images with convolutional neural network.
Severity of white matter lesion (WML) is typically evaluated on magnetic resonance images (MRI), yet the more accessible, faster, and less expensive method is computed tomography (CT). Our objective was to study whether WML can be automatically segmented from CT images using a convolutional neural network (CNN). The second aim was to compare CT segmentation with MRI segmentation. The brain images from the Helsinki University Hospital clinical image archive were systematically screened to make CT-MRI image pairs. Selection criteria for the study were that both CT and MRI images were acquired within 6 weeks. In total, 147 image pairs were included. We used CNN to segment WML from CT images. Training and testing of CNN for CT was performed using 10-fold cross-validation, and the segmentation results were compared with the corresponding segmentations from MRI. A Pearson correlation of 0.94 was obtained between the automatic WML volumes of MRI and CT segmentations. The average Dice similarity index validating the overlap between CT and FLAIR segmentations was 0.68 for the Fazekas 3 group. CNN-based segmentation of CT images may provide a means to evaluate the severity of WML and establish a link between CT WML patterns and the current standard MRI-based visual rating scale.


Percutaneous radiofrequency treatment for refractory anteromedial pain of osteoarthritic knees.
Although severe knee osteoarthritis with refractory pain is commonly treated surgically, this is often not an option for patients with poor health status or unwillingness to undergo major surgery. We examined the efficacy of radiofrequency application to sensory nerves as a novel alternative treatment for refractory knee pain. This study was an open-label, nonrandomized, and controlled study. Patients complaining of refractory anteromedial knee pain associated with radiological osteoarthritis (moderate or severe) were included. They were assigned to one of two groups: those receiving radiofrequency thermocoagulation (N = 18) or those receiving nerve block (N = 17), depending on the time period that they were referred to the clinic. Radiofrequency current or local anesthetics was applied to the medial retinacular nerve and the infrapatellar branch of the saphenous nerve. Western Ontario McMaster Universities osteoarthritis index score, pain visual analog scale (VAS), and patient's global assessment were assessed with a minimum follow-up of 6 months. Radiofrequency treatment significantly decreased knee pain as measured by VAS for 12 weeks compared with the control group. In terms of responders, more patients in the RF group responded to the treatment than in the control group. The differences were statistically significant at 4 weeks, 8 weeks, and 12 weeks in pain VAS. Eight patients (44%) treated with radiofrequency rated excellent or good but only three (18%) in the control group rated good, although the difference was not statistically significant. Some patients were able to benefit substantially from radiofrequency treatment. Even if its effective period is limited, radiofrequency application is a promising treatment to alleviate refractory anteromedial knee pain with osteoarthritis. Further experience and technical improvements are needed to establish its role in the management of knee osteoarthritis.


Living arrangements and the transition to adulthood.
The sharp decline with age in the percent of young adults who live with their parents is usually attributed to other concurrent life-cycle changes in the "transition to adulthood." We investigate this presumption using data tracking high school seniors seven years after graduation. Although marriage and military service strongly reduce residential dependence on parents, other life-cycle changes such as employment and parenthood are only weakly associated with living arrangements and often affect returning home more than leaving. "Leaving home" is often independent of other transition events and should be studied directly to understand recent patterns of family change.


Cross-talk between two organs: how the kidney responds to disruption of acid-base balance by the lung.
Hypoventilation increases PaCO(2) (hypercapnia) and initiates the acid-base disorder known as respiratory acidosis. Hyperventilation decreases PaCO(2) (hypocapnia) and initiates the acid-base disorder known as respiratory alkalosis. The impact on acidity of these primary changes in PaCO(2) is ameliorated by secondary, directional changes in plasma bicarbonate concentration that occur in two stages. Acutely, modest changes in plasma bicarbonate originate from titration of the body's nonbicarbonate buffers. In chronic hypercapnia and hypocapnia, larger changes in plasma bicarbonate occur that reflect adjustments in renal acidification mechanisms. As a result, the amelioration of systemic acidity is more pronounced in the chronic forms of the respiratory acid-base disorders.


[Chronic autoimmune thyroiditis as a predictor and precursor of metaplasia].
In recent years an increase in the incidence of chronic autoimmune thyroiditis (CAT) and its transformation into thyroid cancer (TC) and thyroid lymphoma (TL) have been recorded. The article discusses etiologic and pathogenetic relationship between CAT, TC, TL, PDS and MEN syndromes. Clinical, laboratory, intrascopic and morphological diagnostics of CAT and comorbidities is considered. The algorithm for the choice of the treatment is presented.


A unique phosphorylation-dependent mechanism for the activation of Ca2+/calmodulin-dependent protein kinase type IV/GR.
The activity of the Ca2+/calmodulin-dependent protein kinase IV/Gr (CaMKIV/Gr) is shown to be strictly regulated by phosphorylation of three residues both in vitro and in response to antigen receptor-mediated signaling in lymphocytes. One residue, Thr-200, is indispensable for enhancement of Ca2+/calmodulin-dependent basal activity by CaMKIV/Gr kinase. This event requires Ca2+/calmodulin in the full-length CaMKIV/Gr but is Ca2+/calmodulin-independent when a truncated version of CaMKIV/Gr is used as a substrate (DeltaCaMKIV/Gr1-317 (Delta1-317)). The other two residues, Ser12 and Ser13, are apparently autophosphorylated by the Ca2+/calmodulin-bound CaMKIV/Gr. Phosphorylation of neither Ser12-Ser13 nor Thr312 (the residue in a homologous position to Thr286 of CaMKIIalpha influences the development of Ca2+/calmodulin-independent activity or any other property of CaMKIV/Gr examined. Similarly, removal of the NH2-terminal 20 amino acids has no effect on the activation or function of CaMKIV/Gr. However, mutation of both Ser12 and Ser13 residues to Ala in Delta1-317 completely abrogates activity, while individual substitutions have no effect. These results indicate that the NH2-terminal Ser cluster mediates a novel type of intrasteric inhibition and suggest that three events are required for CaMKIV/Gr activation: 1) Ca2+/calmodulin binding; 2) phosphorylation of the Ca2+/calmodulin-bound enzyme on Thr200 by a Ca2+/calmodulin-dependent protein kinase kinase; and 3) autophosphorylation of Ser12-Ser13. This three-step requirement is unique among the multifunctional Ca2+/calmodulin-dependent kinases.


First norovirus outbreaks associated with consumption of green seaweed (Enteromorpha spp.) in South Korea.
In February 2012, an outbreak of gastroenteritis was reported in school A; a successive outbreak was reported at school B. A retrospective cohort study conducted in school A showed that seasoned green seaweed with radishes (relative risk 7·9, 95% confidence interval 1·1-56·2) was significantly associated with illness. Similarly, a case-control study of students at school B showed that cases were 5·1 (95% confidence interval 1·1-24·8) times more likely to have eaten seasoned green seaweed with pears. Multiple norovirus genotypes were detected in samples from students in schools A and B. Norovirus GII.6 isolated from schools A and B were phylogenetically indistinguishable. Green seaweed was supplied by company X, and norovirus GII.4 was isolated from samples of green seaweed. Green seaweed was assumed to be linked to these outbreaks. To our knowledge, this is the first reported norovirus outbreak associated with green seaweed.


Co-developed implementation guidelines to maximize acceptability, feasibility, and usability of mobile phone supervision in Kenya.
Opportunities exist to leverage mobile phones to replace or supplement in-person supervision of lay counselors. However, contextual variables, such as network connectivity and provider preferences, must be considered. Using an iterative and mixed methods approach, we co-developed implementation guidelines to support the implementation of mobile phone supervision with lay counselors and supervisors delivering a culturally adapted trauma-focused cognitive behavioral therapy in Western Kenya. Guidelines were shared and discussed with lay counselors in educational outreach visits led by supervisors. We evaluated the impact of guidelines and outreach on the acceptability, feasibility, and usability of mobile phone supervision. Guidelines were associated with significant improvements in acceptability and usability of mobile phone supervision. There was no evidence of a significant difference in feasibility. Qualitative interviews with lay counselors and supervisors contextualized how guidelines impacted acceptability and feasibility - by setting expectations for mobile phone supervision, emphasizing importance, increasing comfort, and sharing strategies to improve mobile phone supervision. Introducing and discussing co-developed implementation guidelines significantly improved the acceptability and usability of mobile phone supervision. This approach may provide a flexible and scalable model to address challenges with implementing evidence-based practices and implementation strategies in lower-resourced areas.


Inhibition of streptomycin-dependent mutation in Escherichia coli on the lytic growth of bacteriophage lambda.
Spontaneous streptomycin-dependent mutants (StrDA) were isolated from Escherichia coli C600. On C600 StrD, the lytic growth of phage lambda Nam and lambda cI857 was inhibited. After E. coli lysogenic strain 1.1485 (lambda cI857) mutated to StrDA, induction of lambda was decreased greatly. On StrDA of E. coli strains C600 and 1.1485 (lambda cI857), the plating efficiencies and burst sizes of phage T4 and T7 remained normal. Since StrDA is a mutation in the structural gene for ribosomal protein S12, the results obtained in the present study suggest that ribosomes of the StrDA mutants inhibit the lytic growth of lambda phage. The possibility that StrDA ribosomes inhibit the expression of lambda N gene is discussed based on the comparison of the genetic background of lambda cI857 and lambda Nam.


Electroconvulsive Therapy for Parkinson's Disease: A Systematic Review and Meta-Analysis.
Electroconvulsive therapy (ECT) is a well-established treatment for psychiatric disorders, including depression and psychosis. ECT has been reported to be effective in treating such psychiatric symptoms in patients with Parkinson's disease (PD) and has been also reported to be effective in treating motor symptoms. The aim of the study is to summarize previous clinical studies investigating the efficacy of ECT for symptoms in patients with PD. A systematic review and meta-analysis of any study designs assessing motor and/or non-motor symptoms in patients with PD before and after ECT. Co-primary outcomes were set as motor manifestations assessed using the Unified Parkinson's Disease Rating Scale or other rating scales, and non-motor symptoms included depression and psychosis. Secondary outcomes were wearing-off phenomenon and cognitive function. The impact of ECT on those symptoms was examined by comparing the severity of the symptoms before and after ECT using a random effect model and was expressed in standardized mean difference. Of 1219 identified citations, 14 studies (n = 129; 1 randomized controlled study, 9 prospective observational studies, and 4 retrospective studies) were analyzed. The findings were as follows: ECT significantly improved motor manifestations in patients with PD, and the improvement was significant in the subpopulation without psychiatric symptoms; ECT significantly improved depression and psychosis; and ECT significantly relieved wearing-off phenomenon and did not worsen cognitive functioning. The current meta-analysis suggests the potential benefit of ECT on motor and non-motor symptoms in presumably complicated and difficult-to-treat subgroups. © 2020 International Parkinson and Movement Disorder Society.


DNA adductomics.
Systems toxicology is a broad-based approach to describe many of the toxicological features that occur within a living system under stress or subjected to exogenous or endogenous exposures. The ultimate goal is to capture an overview of all exposures and the ensuing biological responses of the body. The term exposome has been employed to refer to the totality of all exposures, and systems toxicology investigates how the exposome influences health effects and consequences of exposures over a lifetime. The tools to advance systems toxicology include high-throughput transcriptomics, proteomics, metabolomics, and adductomics, which is still in its infancy. A well-established methodology for the comprehensive measurement of DNA damage resulting from every day exposures is not fully developed. During the past several decades, the (32)P-postlabeling technique has been employed to screen the damage to DNA induced by multiple classes of genotoxicants; however, more robust, specific, and quantitative methods have been sought to identify and quantify DNA adducts. Although triple quadrupole and ion trap mass spectrometry, particularly when using multistage scanning (LC-MS(n)), have shown promise in the field of DNA adductomics, it is anticipated that high-resolution and accurate-mass LC-MS(n) instrumentation will play a major role in assessing global DNA damage. Targeted adductomics should also benefit greatly from improved triple quadrupole technology. Once the analytical MS methods are fully mature, DNA adductomics along with other -omics tools will contribute greatly to the field of systems toxicology.


Color-by-blue display using blue quantum dot light-emitting diodes and green/red color converting phosphors.
We report a novel full-color display based on the generation of full-color by a highly efficient blue QD-LED light approach, or so called color-by-blue QD-LED display. This newly proposed color-by-blue QD-LED display combines a blue CdZnS/ZnS QD-LED blue subpixel and excitation source with front-emitting green/red phosphor subpixels. It is carefully estimated that the detailed display characteristics as well as full color-conversion and reasonable device efficiency of blue, green, and red satisfy the minimum requirements for display application. Also, we would like to emphasize that the proposed blue, green, and red device shows maximum luminance of 1570, 12920, and 3120 cd/m², respectively, luminous efficiency of 1.5, 12.1, and 2.5 cd/A, respectively, and external quantum efficiency of 6.8, 2.8, and 2.0%, respectively. It is expected that full color generation by color-by-blue QD-LED will lead to further technological advancements in the area of efficient and facile display applications.


Phenethyl isothiocyanate reduces breast cancer stem cell-like properties by epigenetic reactivation of CDH1.
Breast cancer is the most frequently diagnosed malignancy and leading cause of cancer-related deaths among women worldwide. Tumor recurrence, or metastasis, is caused by cancer stem cells and has a dismal prognosis for breast cancer patients. Thus, targeting breast cancer stem cells (BCSCs) for eradication is a potential method to improve clinical outcomes. Phenethyl isothiocyanate (PEITC) is a novel epigenetic regulator derived from cruciferous vegetables that has marked antitumor effects. However, the exact mechanism of these antitumor effects by PEITC is unknown. As breast cancer progresses, a tumor suppressor in the breast, cadherin 1 (CDH1), is silenced by hypermethylation of the promoter region, further promoting the stem cell-like properties of cancer. Herein, the ability of PEITC to reduce BCSC-like properties by epigenetic reactivation of CDH1 was investigated by multiple analyses such as MTT, colony formation and sphere formation assays, methylation-specific PCR, western blot analysis, Co-IP and qPCR. It was revealed that PEITC inhibited colony and mammosphere formation and decreased the expression of protein markers associated with BCSC-like properties via epigenetic reactivation of CDH1. Further exploration of this mechanism revealed inhibitory effects of PEITC on DNMTs and HDACs, which play a pivotal role in demethylating the hypermethylated CDH1 promoter region. Reactivated CDH1 suppressed the Wnt/β-catenin pathway which confers BCSC-properties in breast cancer cells. These findings suggest a novel method to eradicate BCSCs from breast cancer patients.


The pharmacokinetics of flurbiprofen in younger and elderly patients with rheumatoid arthritis.
The pharmacokinetics of flurbiprofen (Ansaid Tablets, Upjohn Company of Canada, Don Mills, Ontario) were evaluated in both younger (40 to 60 years) and elderly (65 to 83 years) rheumatoid arthritic patients after both a 100-mg single-dose administration and at steady state during a 100-mg twice-a-day dosage regimen. Both flurbiprofen plasma concentration-time profiles and the urinary excretion of flurbiprofen and its major metabolites were evaluated. The results indicate that the pharmacokinetics of flurbiprofen are linear in both age groups based on only minor changes between single-dose and steady-state parameter determinations and the agreement between calculated and predicted accumulation values in plasma concentrations. Only minor differences in the pharmacokinetic parameters were observed between the younger and elderly patients. Only free flurbiprofen clearance was found to have a significant but variable correlation to patient age. The effect of flurbiprofen on the urinary excretion of two prostaglandins were also evaluated throughout this study. In both age groups, the maximum decrease in urinary excretion was observed after the first dose, and this effect was maintained throughout the remainder of the study. Percent decreases from baseline in urinary excretion during drug administration were similar for both age groups. Similar side-effect profiles were observed between age groups.


The mechanism of lncRNA TALNEC2 regulating miR-19a-3p/JNK to alleviate cerebral ischemia injury in rats with acute cerebral infarction.
Ischemia-reperfusion (I-R) is renowned as a key approach in recovery related to cerebral infarction and further promotes succeeding infarction development. This study investigated the fundamental molecular function of the TALNEC2 in the pathogenesis of cerebral infarction to provide insights on the potential novel therapeutic agents in cerebral infarction. RT-qPCR measured expression of TALNEC2 and JNK in human neural cell line SH-SY5Y. Cell transfection upregulated or silenced the genes with MTT assay examining cell viability. RT-qPCR detected cell death in the apoptosis biomarker caspase-3, inflammation in the biomarkers C-reactive protein (CRP) and IL-6 and verified cell proliferation via the ki67 and PCNA markers. Luciferase assay was performed to see the luciferase activity and western blotting determined the protein expression of JNK in proliferation, inflammation. The results demonstrated that TALNEC2 was highly expressed after OGD/R treatment in nerve cells after cerebral infarction. In addition, TALNEC2 silencing prevented apoptosis and inflammation of nerve cells after cerebral infarction. TALNEC2 directly interacted with miR-19a-3p to regulate JNK protein expression. Lastly, miR-19a-3p inhibitor abolished the protective effect of si-TALNEC2 against OGD/R induced damage in vitro. In summary, this study has demonstrated that TALNEC2 is a positive moderator for pathogenesis of cerebral infarction. Furthermore, our conclusions provide further insights on the interplay among TALNEC2, miR-19a-3p and JNK in cerebral infarction. It has demonstrated herein that TALNEC2 positively modulates JNK post-transcriptional expression through miR-19a-3p sponging in cerebral Infarction offering a novel therapy target for cerebral infarction.


Induction of ErbB-2/neu-specific protective and therapeutic antitumor immunity using genetically modified dendritic cells: enhanced efficacy by cotransduction of gene encoding IL-12.
Overexpression of ErbB-2/neu occurs in 20-30% of patients with breast cancer and indicates a poor prognosis. The presence of a detectable immune response to ErbB-2/neu in some patients suggests that this oncogene may be a useful target for vaccine therapy. We evaluated whether genetic immunization using dendritic cells (DC) transduced ex vivo with an adenovirus expressing the ErbB-2/neu gene (AdNeuTK) could induce protective and therapeutic immunity against a breast tumor cell line overexpressing ErbB-2/neu. Subcutaneous (s.c.) immunization with the DC vaccine elicited protective immunity in an average of 60% of animals. CTL analysis demonstrated specific cytotoxic activity against breast tumor cells, as well as syngeneic fibroblasts transduced with AdNeuTK. In vivo depletion studies demonstrated both CD4+ and CD8+ T cells were required. In a therapeutic setting, immunization with the DC vaccines could cure mice with pre-established tumors and efficacy was further enhanced by cotransducing DCs with a vector expressing murine IL-12 (AdmIL-12). These studies support DC vaccines as a therapeutic strategy for human breast cancer, while emphasizing the importance of optimizing an immune response by combining tumor antigen presentation with immunostimulatory cytokines.


Epidermal growth factor enemas with oral mesalamine for mild-to-moderate left-sided ulcerative colitis or proctitis.
Epidermal growth factor (EGF) is a potent mitogenic peptide produced by salivary glands. We examined whether EGF enemas are an effective treatment for active left-sided ulcerative colitis and ulceration limited to the rectum (proctitis). In a randomized, double-blind clinical trial conducted at Leicester Royal Infirmary, 12 patients with mild-to-moderate left-sided ulcerative colitis received daily enemas of 5 microg of EGF in 100 ml of an inert carrier and 12 received daily enemas with carrier alone for 14 days. All also began to receive 1.2 g of oral mesalamine per day or had their dose increased by 1.2 g per day. Patients were assessed clinically at 0, 2, 4, and 12 weeks and by sigmoidoscopy and biopsy at 0, 2, and 4 weeks. The primary end point was disease remission (defined by a St. Marks score of 4 or less without sigmoidoscopic evidence of inflammation) at two weeks. Secondary end points were clinically significant improvements in disease activity (defined by a decrease of more than 3 points in the St. Marks score or the ulcerative colitis disease-activity index) at two and four weeks. Analyses were performed according to the intention-to-treat principle. After two weeks, 10 of the 12 patients given EGF enemas were in remission, as compared with 1 of 12 in the control group (83 percent vs. 8 percent, P<0.001). At the 2-week assessment, disease-activity scores, sigmoidoscopic score, and histologic scores were all significantly better in the EGF group than in the placebo group (P<0.01 for all comparisons), and this benefit was maintained at 4 weeks and at 12 weeks. This study provides preliminary data suggesting that EGF enemas are an effective treatment for active left-sided ulcerative colitis.


Influence of nutrient level on methylmercury content in water spinach.
Widely consumed vegetables are often cultivated in sewage waters with high nutrient levels. They can contain high levels of methylmercury (MeHg), because they can form MeHg from inorganic Hg in their young shoots. We determined whether the MeHg uptake and the MeHg formation in the shoots of water spinach (Ipomoea aquatica) were affected by the presence of a high nutrient level in the growth medium. Water spinach shoots were rooted and pretreated in growth medium containing 7% (low) or 70% (high) Hoagland nutrient solution; thereafter, the plants were treated with either 0.02 microM MeHg or 0.2 microM HgCl2 for 3 d. Half the plants were then analyzed for total Hg and MeHg. The remaining plants were transferred to mercury-free medium with low or high nutrient levels and posttreated for 3 days before analysis of total Hg and MeHg in order to measure MeHg formation in the absence of external Hg. The results indicate that nutrient level did not influence MeHg uptake, but that a high nutrient level reduced the distribution of MeHg to the shoots 2.7-fold versus low nutrient level. After treatment with HgCl2, MeHg was found in roots and new shoots but not in old shoots. The MeHg:total-Hg ratio was higher in new shoots than in roots, being 13 times higher at high versus low nutrient levels. Thus, MeHg formation was the same in new shoots independent of inorganic Hg concentration, since the total Hg level decreased at a high nutrient level.


Encapsulation of functional organic compounds in nanoglass for optically anisotropic coatings.
A novel approach is presented for the encapsulation of organic functional molecules between two sheets of 1 nm thin silicate layers, which like glass are transparent and chemically stable. An ordered heterostructure with organic interlayers strictly alternating with osmotically swelling sodium interlayers can be spontaneously delaminated into double stacks with the organic interlayers sandwiched between two silicate layers. The double stacks show high aspect ratios of >1000 (typical lateral extension 5000 nm, thickness 4.5 nm). This newly developed technique can be used to mask hydrophobic functional molecules and render them completely dispersible in water. The combination of the structural anisotropy of the silicate layers and a preferred orientation of molecules confined in the interlayer space allows polymer nanocomposite films to be cast with a well-defined orientation of the encapsulated molecules, thus rendering the optical properties of the nanocoatings anisotropic.


Robot-assisted total mesorectal excision: is there a learning curve?
Laparoscopic total mesorectal excision (TME) is associated with a steep learning curve, but the learning curve for robotic TME is unknown. This study aimed to evaluate the learning curve for robotic TME. Between November 2004 and April 2009, 80 patients underwent robotic TME performed by a single surgeon. The operative experience was divided into two groups: group 1 (the first 40 cases) and group 2 (the subsequent 40 cases). Patient demographics, operative characteristics, and morbidities were compared. The two patient populations selected did not differ statistically in age, body mass index (BMI), preoperative risk assessment, stage, preoperative chemoradiotherapy, or tumor location. The mean operative times in group 1 (310 min) and group 2 (297 min) were similar (p = 0.55), and the mean robotic TME time did not differ between the two groups (60 vs. 64 min; p = 0.65). In addition, the operative times did not improve during the course of the study. There were no differences in EBL, margin status, or number of lymph nodes harvested. Furthermore, there were no differences in conversion rate, time to resumption of diet, length of hospital stay, or postoperative complications. Robot-assisted TME may attenuate the learning curve for laparoscopic rectal cancer resection. Further studies are necessary to establish the role of robotic surgery in minimally invasive rectal operations.


Na+-K+-ATPase activity in medullary thick ascending limb during short-term anoxia.
Na+-K+-ATPase activity was measured in a suspension of rabbit medullary thick ascending limb tubules under oxygenated and anoxic conditions. Oxygenated, K-depleted tubules rapidly take up added extracellular potassium accompanied by a simultaneous increase in oxygen consumption. The ATP/O2 ratio was 12.5 +/- 0.7, suggesting a tight coupling between oxidative metabolism (6 ATP/O2) and Na+-K+-ATPase activity (2 K/ATP). On reaching anoxia, the tubules released potassium into the medium, but this rate was accelerated by the addition of ouabain, which indicated that the Na+-K+-ATPase was still operative in anoxia. Because 10 min of anoxia led to only a 15.7% decline in potassium content, a new steady state of potassium uptake and leakage must be reached during anoxia. Anaerobic metabolism maintained 73% of cellular ATP during 10 min of anoxia. Exposure of anoxic tubules to iodoacetate produced a 57% decline in ATP levels and a 33% decline in potassium content, which indicated that glycolysis is an important pathway in supplying energy during anaerobiosis.


[Use of the restriction analysis in the quality control of the strain of Cuban biolarvicide BACTIVEC].
The genomic characterization of the strain used in manufacturing biolarvicide BACTIVET was shown in this paper. Digestion patterns of Bacillus thuringiensis reference strain 266 from the collection of Pushkin Institute located in former Leningrad and of two strains isolated from 2 batches of biolarvicide were compared. Chromosomal DNA digestion was performed by enzymes Hindlll and EcoRI. The same restriction pattern was observed in all cases, which is indicative of genetic stability of the active ingredient of biolarvicide BACTIVET.


It is safe to use minimal restrictions following posterior approach total hip arthroplasty: results from a large cohort study.
To prevent early postoperative dislocation following a total hip arthroplasty (THA) procedure, patients must adhere to restrictions. Restrictive protocols are common if THA surgery is performed using the posterior approach, but scientific evidence form larger studies that supports these restrictions are scarce. In this large cohort study we compare the <90-days dislocation rate between patients receiving posterior approach THA managed with minimal versus extensive restrictions. Prospective cohort (n = 1049) of consecutive elective primary hip replacement surgery procedures (September 2014-July 2017) managed with minimal postoperative restrictions. Hospital charts were prospectively reviewed for patient demographics, risk factors and any hip dislocation. Control (n = 1102) consecutive primary elective THAs (January 2011-August 2014) managed with a traditional restrictive protocol. A posterior surgical approach was used in all procedures. Minimal restrictions group: 17 dislocations <90 days (1.6%); Restricted group: 28 (2.5%), chi-square p = 0.1. Testing the hypothesis of inferiority by a minimum of 1% increase in ⩽90 days dislocation risk: p = 0.14 (test for difference) and p < 0.001 (non-inferiority test), allowing us to discard the null hypothesis (absolute increase in risk of ⩾1% with minimal restrictions). The proportion of surgeries performed with a femoral head size ⩾32 mm was higher in the minimal restrictions group. Patients can be managed safely with minimal restrictions following posterior approach THA if combined with frequent use of larger femoral heads.


Anti-proteinuric effects of combination therapy with enalapril and losartan in patients with nephropathy due to type 2 diabetes.
The benefits of angiotensin-converting enzyme inhibitors and angiotensin II (ATII) receptor antagonist therapy of diabetic nephropathy (DNP) are thought to be largely the result of attenuation of ATII effects on proteinuria. The aim of the study was to ascertain whether there is the additive anti-proteinuric effect of enalapril plus losartan in DNP. Twenty-two patients with DNP were studied. Patients were randomly assigned to enalapril 10 mg/day (11 patients) or losartan 50 mg/day (11 patients) administered in a single oral dose in the morning for 12 weeks. and then, in 10 patients (five patients from enalapril group and five patients from losartan group), combination therapy (10 mg/day enalapril and 50 mg/day losartan) was started and continued for 12 weeks. In 12 patients, initial drugs dosages were doubled (six patients 20 mg/day enalapril and six patients 100 mg/day losartan), and monotherapy was continued for 12 weeks. Blood pressure and proteinuria were measured before and after therapy. Adverse effects were recorded at every visit. Proteinuria decreased by 33% with enalapril and losartan administered alone (p < 0.05). Co-administration of enalapril and losartan decreased proteinuria by a greater extent compared with enalapril and losartan administered alone (51%, p<0.05). This proteinuria level was significantly lower than the proteinuria level of 12 weeks therapy with enalapril and losartan alone. The decrease of proteinuria was 37% in double-dose monotherapy group (p < 0.05). Reduction of mean arterial blood pressure (MAP) in co-administration of enalapril and losartan was higher than enalapril and losartan administered alone (p < 0.05). Combination of enalapril and losartan decreased proteinuria and MAP by a greater extent compared with enalapril and losartan administered alone. We have found that proteinuria reduction induced by combined therapy is maintained throughout short-term follow-up; a greater anti-proteinuric response was observed in the patients with DNP.


Percutaneous mitral valve repair for mitral regurgitation.
Mitral regurgitation (MR) associated with, ischemic, and degenerative (prolapse) disease, contributes to left ventricular (LV) dysfunction due to remodeling, and LV dilation, resulting in worsening of MR. Mitral valve (MV) surgical repair has provided improvement in survival, LV function and symptoms, especially when performed early. Surgical repair is complex, due to diverse etiologies and has significant complications. The Society for Thoracic Surgery database shows that operative mortality for a 1st repair is 2% and for re-do repair is 4 times that. Cardiopulmonary bypass and cardiac arrest are required. The attendant morbidity prolongs hospitalization and recovery. Alfieri simplified mitral repair using an edge-to-edge technique which subsequently has been shown to be effective for multiple etiologies of MR. The MV leaflers are typically brought together by a central suture producing a double orifice MV without stenosis. Umana reported that MR decreased from grade 3.6 +/- 0.5 to 0.8 +/- 0.4 (P < 0.0001) and LV ejection fraction increased from 33 +/- 13% to 45 +/- 11% (P = 0.0156). In 121 patients, Maisano reported freedom from re-operation of 95 +/- 4.8% with up to 6 year follow-up. Oz developed a MV "grasper" that is directly placed via a left ventriculotomy and coapts both leaflets which are then fastened by a graduated spiral screw. An in-vitro model using explanted human valves showed significant reduction in MR and in canine studies, animals followed by serial echo had persistent MV coaptation. At 12 weeks the device was endothelialized. These promising results have paved the way for a percutaneous or minimally invasive-off pump mitral repair. Evalve has developed catheter-based technology, which, by apposing the edges of a regurgitant MV, results in edge-to-edge repair. Release of the device is done after echo and fluoroscopic evaluation under normal loading conditions. If the desired effect is not produced the device can be repositioned or retrieved. Animal studies show excellent healing, with incorporation of the device into the leaflets at 6-10 weeks with persistent coaptation. Another percutaneous approach has been to utilize the proximity of the coronary sinus (CS) to the mitral annulus (MA). Placement of a self-compressing device in the CS along the region of the posterior MA has, in canine models, reduced MR and addresses the issues of MA dilation and its contribution to MR. Ongoing studies are underway for both techniques.


Chlorogenic acid versus amaranth's caffeoylisocitric acid - Gut microbial degradation of caffeic acid derivatives.
The almost forgotten crop amaranth has gained renewed interest in recent years due to its immense nutritive potential. Health beneficial effects of certain plants are often attributed to secondary plant metabolites such as phenolic compounds. As these compounds undergo significant metabolism after consumption and are in most cases not absorbed very well, it is important to gain knowledge about absorption, biotransformation, and further metabolism in the human body. Whilst being hardly found in other edible plants, caffeoylisocitric acid represents the most abundant low molecular weight phenolic compound in many leafy amaranth species. Given that this may be a potentially bioactive compound, gastrointestinal microbial degradation of this substance was investigated in the present study by performing in vitro fermentation tests using three different fecal samples as inocula. The (phenolic) metabolites were analyzed using high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS). Furthermore, quantitative polymerase chain reaction (qPCR) analyses were carried out to study the influence on the microbiome and its composition. The in vitro fermentations led to different metabolite profiles depending on the specific donor. For example, the metabolite 3-(4-hydroxyphenyl)propionic acid was observed in one fermentation as the main metabolite, whereas 3-(3-hydroxyphenyl)propionic acid was identified in the other fermentations as important. A significant change in selected microorganisms of the gut microbiota however was not detected. In conclusion, caffeoylisocitric acid from amaranth, which is a source of several esterified phenolic acids in addition to chlorogenic acid, can be metabolized by the human gut microbiota, but the metabolites produced vary between individuals.


Role of adherence in interleukin-8 induction in Helicobacter pylori-associated gastritis.
Active Helicobacter pylori-associated gastritis is characterized by a dense mucosal infiltration with granulocytes. Since H. pylori is noninvasive, secondary signals must induce the accumulation of granulocytes. Interleukin-8 (IL-8) has been shown to play a key role in this event. Using competitive reverse transcriptase-PCR on mRNA from gastric biopsies, we could show a clear correlation between the amount of IL-8 transcripts and the activity of H. pylori gastritis. Due to the inability of the bacterium to invade host cells, the epithelial layer is a potential candidate as an IL-8 source. To study the mechanism of IL-8 induction, established gastric carcinoma epithelial cell lines (AGS and Kato III) and well-defined H. pylori strains were used in a modified in vitro system. The experimental design enabled us to prevent direct contact of bacteria with epithelial cells by use of a filter membrane which did not block secreted bacterial products crossing the membrane. The data clearly showed that the direct contact of the bacterial cell with the epithelial cell is necessary for optimal IL-8 production because not only live bacteria, but also metabolically inactive bacteria, increased IL-8 secretion. Neither purified lipopolysaccharide nor water-soluble protein fractions of H. pylori NCTC 11637 and Tx30a nor the cytotoxin of H. pylori was able to increase IL-8 production significantly by the epithelial cells used. Furthermore, preparations of total membrane and outer membrane proteins of H. pylori were not able to stimulate IL-8 release in vitro. Accumulatively, these results imply that active metabolism is not necessary for stimulation as long as there is an intact membrane aiding the presentation of a stimulating membrane complex or aggregate on the surface of the bacteria. From these results, we conclude that whole bacteria and their direct contact with epithelial cells may be critical for IL-8 induction in vivo.


Haloadaptative Responses of Aspergillus sydowii to Extreme Water Deprivation: Morphology, Compatible Solutes, and Oxidative Stress at NaCl Saturation.
Water activity (aw) is critical for microbial growth, as it is severely restricted at aw < 0.90. Saturating NaCl concentrations (~5.0 M) induce extreme water deprivation (aw ≅ 0.75) and cellular stress responses. Halophilic fungi have cellular adaptations that enable osmotic balance and ionic/oxidative stress prevention to grow at high salinity. Here we studied the morphology, osmolyte synthesis, and oxidative stress defenses of the halophile Aspergillus sydowii EXF-12860 at 1.0 M and 5.13 M NaCl. Colony growth, pigmentation, exudate, and spore production were inhibited at NaCl-saturated media. Additionally, hyphae showed unpolarized growth, lower diameter, and increased septation, multicellularity and branching compared to optimal NaCl concentration. Trehalose, mannitol, arabitol, erythritol, and glycerol were produced in the presence of both 1.0 M and 5.13 M NaCl. Exposing A. sydowii cells to 5.13 M NaCl resulted in oxidative stress evidenced by an increase in antioxidant enzymes and lipid peroxidation biomarkers. Also, genes involved in cellular antioxidant defense systems were upregulated. This is the most comprehensive study that investigates the micromorphology and the adaptative cellular response of different non-enzymatic and enzymatic oxidative stress biomarkers in halophilic filamentous fungi.


Some effects of the formamidine pesticide chlordimeform on the behavior of mice.
Three experiments were performed to determine the behavioral effects of acutely administered chlordimeform (1-56 mg/kg, i.p.) to male mice. In Experiment I, dosages of 30 and 56 mg/kg decreased the amount of time spent investigating a female conspecific. In Experiment II, large dosages of chlordimeform (30 and 56 mg/kg) decreased vertically directed activity (rearing) to a proportionately greater extent than horizontally directed activity (ambulation). In Experiment III, chlordimeform produced conditioned flavor aversions that were intermediate at 10 mg/kg and maximal at 30 mg/kg. In an additional experiment (Experiment IV), the LD50 for chlordimeform was determined to be 95.6 mg/kg. Behavioral effects were thus obtained at dosages that were approximately 10 to 30% of the LD50.


Suppressed histamine release from rat peritoneal mast cells by ultraviolet B irradiation: decreased diacylglycerol formation as a possible mechanism.
This study was designed to investigate the effect of ultraviolet B (UVB) irradiation on mast cell functions. Purified mast cells obtained from rat peritoneal cavity were irradiated with UVB and subsequently exposed to a degranulator, compound 48/80, or the calcium ionophore A-23187. The amount of histamine released from mast cells measured by the enzyme isotopic assay was significantly decreased by UVB irradiation (100-400 mJ/cm2). Within this dose range, UVB alone was not cytotoxic to the cells because it did not induce histamine release. The suppression was observed when mast cells were subjected to degranulation without intervals after UVB irradiation, and even after 5 h postirradiation. The wavelength of 300 nm from a monochromatic light source showed the maximum effect. When mast cells prelabeled with [3H]arachidonate were irradiated and challenged by compound 48/80, label accumulation in diacylglycerol produced by the phosphatidylinositol cycle was considerably decreased by UVB irradiation. From these results, we hypothesize that, within an adequate irradiation dose, UVB irradiation suppresses histamine release from mast cells, probably by causing noncytotoxic damage to the membrane phospholipid metabolism, which is tied to the degranulation mechanisms.


Dynamic analysis of a rhythmic neural circuit in the leech Hirudo medicinalis.
1. The results of perturbation experiments demonstrate the functional diversity of the interneurons (HN cells) that generate heartbeat in the medicinal leech. 2. HN cells were individually stimulated by single current pulses. The induced activity of HN cells in the first four ganglia (cell pairs HN(1)-HN(4)) reset the rhythm of the interneuron network; induced activity of those in the fifth through seventh ganglia (cell pairs HN(5)-HN(7)) did not. 3. Cells HN(1)-HN(4) can entrain every other interneuron of the network; cells HN(5)-HN(7) cannot. 4. Thus the HN interneuron network includes two distinct subsets: cells HN(1)-HN(4) form the network's timing oscillator; cells HN(5)-HN(7), driven by the timing oscillator, force one of the two coordination states on the heart motor neurons. 5. In general the dynamic behavior of the heart interneuron network was predictable given the web of identified synapses between HN cells. Nevertheless, the unexpected capacity of cells HN(3) and HN(4) to entrain the network shows that there are functional connections still to be found. Burst termination experiments suggest that cells HN(3) and HN(4) inhibit directly the more rostral HN cells. 6. The timing oscillation seems to arise from a balance between the endogenous polarization rhythms of interneurons HN(1)-HN(4) and selective reciprocal inhibition between these same cells.


Electrophysiology of the insect dorsal ocellus. I. Origin of the components of the electroretinogram.
Dorsal ocelli are small cup-like organs containing a layer of photoreceptor cells, the short axons of which synapse at the base of the cup with dendritic terminals of ocellar nerve fibers. The ocellar ERG of dragonflies, recorded from the surface of the receptor cell layer and from the long lateral ocellar nerve, contains four components. Component 1 is a depolarizing sensory generator potential which originates in the distal ends of the receptor cells and evokes component 2. Component 2 is believed to be a depolarizing response of the receptor axons. It evokes a hyperpolarizing postsynaptic potential, component 3, which originates in the dendritic terminals of the ocellar nerve fibers. Ocellar nerve fibers in dragonflies are spontaneously active, discharging afferent nerve impulses (component 4) in the dark-adapted state. Component 3 inhibits this discharge. The ERG of the cockroach ocellus is similar. The main differences are that component 3 is not as conspicuous as in the dragonflies and that in most cases ocellar nerve impulses appear only as a brief burst at "off." In one preparation a spontaneous discharge of nerve impulses was observed. As in the dragonflies, this was inhibited by illumination.


The consequences of hospital autonomization in Colombia: a transaction cost economics analysis.
Granting autonomy to public hospitals in developing countries has been common over recent decades, and implies a shift from hierarchical to contract-based relationships with health authorities. Theory on transactions costs in contractual relationships suggests they stem from relationship-specific investments and contract incompleteness. Transaction cost economics argues that the parties involved in exchanges seek to reduce transaction costs. The objective of this research was to analyse the relationships observed between purchasers and the 22 public hospitals of the city of Bogota, Colombia, in order to understand the role of relationship-specific investments and contract incompleteness as sources of transaction costs, through a largely qualitative study. We found that contract-based relationships showed relevant transaction costs associated mainly with contract incompleteness, not with relationship-specific investments. Regarding relationships between insurers and local hospitals for primary care services, compulsory contracting regulations locked-in the parties to the contracts. For high-complexity services (e.g. inpatient care), no restrictions applied and relationships suggested transaction-cost minimizing behaviour. Contract incompleteness was found to be a source of transaction costs on its own. We conclude that transaction costs seemed to play a key role in contract-based relationships, and contract incompleteness by itself appeared to be a source of transaction costs. The same findings are likely in other contexts because of difficulties in defining, observing and verifying the contracted products and the underlying information asymmetries. The role of compulsory contracting might be context-specific, although it is likely to emerge in other settings due to the safety-net role of public hospitals.


Poisson-Boltzmann continuum-solvation models: applications to pH-dependent properties of biomolecules.
All molecules can be viewed as either discrete or continuous assemblies of electric charges, and electrostatics plays a major role in intermolecular and intramolecular interactions. Moreover, charge distribution within molecules may fluctuate due to the presence of ionizable groups capable of exchanging protons with the environment, leading to pH-dependence of phenomena involving such molecules. Electrostatic aspects of complex shapes and environments of biological molecules, in vitro and in vivo, are relatively well amenable to treatment by Poisson-Boltzmann models, which are attractive in that they possess a clear physical meaning, and can be readily solved by several mathematically sound methods. Here we describe applications of these models to obtain valuable insights into some biologically important pH-dependent properties of biomolecules, such as stability, binding of ligands (including potential drugs), enzymatic activity, conformational transitions, membrane transport and viral entry.


CD95/Fas suppresses NF-κB activation through recruitment of KPC2 in a CD95L/FasL-independent mechanism.
CD95 expression is preserved in triple-negative breast cancers (TNBCs), and CD95 loss in these cells triggers the induction of a pro-inflammatory program, promoting the recruitment of cytotoxic NK cells impairing tumor growth. Herein, we identify a novel interaction partner of CD95, Kip1 ubiquitination-promoting complex protein 2 (KPC2), using an unbiased proteomic approach. Independently of CD95L, CD95/KPC2 interaction contributes to the partial degradation of p105 (NF-κB1) and the subsequent generation of p50 homodimers, which transcriptionally represses NF-κB-driven gene expression. Mechanistically, KPC2 interacts with the C-terminal region of CD95 and serves as an adaptor to recruit RelA (p65) and KPC1, which acts as E3 ubiquitin-protein ligase promoting the degradation of p105 into p50. Loss of CD95 in TNBC cells releases KPC2, limiting the formation of the NF-κB inhibitory homodimer complex (p50/p50), promoting NF-κB activation and the production of pro-inflammatory cytokines, which might contribute to remodeling the immune landscape in TNBC cells.


Regulation of hepatic glucose production during exercise in humans: role of sympathoadrenergic activity.
To investigate the role of sympathoadrenergic activity on glucose production (Ra) during exercise, eight healthy males bicycled 20 min at 41 +/- 2 and 74 +/- 4% maximal O2 uptake (VO2max; mean +/- SE) either without (control; Co) or with blockade of sympathetic nerve activity to liver and adrenal medulla by local anesthesia of the celiac ganglion (Bl). Epinephrine (Epi) was in some experiments infused during blockade to match (normal Epi) or exceed (high Epi) Epi levels during Co. A constant infusion of somatostatin and glucagon was given before and during exercise. At rest, insulin was infused at a rate maintaining euglycemia. During intense exercise, insulin infusion was halved to mimic physiological conditions. During exercise, Ra increased in Co from 14.4 +/- 1.0 to 27.8 +/- 3.0 mumol.min-1.kg-1 (41% VO2max) and to 42.3 +/- 5.2 (74% VO2max; P < 0.05). At 41% VO2max, plasma glucose decreased, whereas it increased during 74% VO2max. Ra was not influenced by Bl. In high Epi, Ra rose more markedly compared with control (P < 0.05), and plasma glucose did not fall during mild exercise and increased more during intense exercise (P < 0.05). Free fatty acid and glycerol concentrations were always lower during exercise with than without celiac blockade. We conclude that high physiological concentrations of Epi can enhance Ra in exercising humans, but normally Epi is not a major stimulus. The study suggests that neither sympathetic liver nerve activity is a major stimulus for Ra during exercise. The Ra response is enhanced by a decrease in insulin and probably by unknown stimuli.(ABSTRACT TRUNCATED AT 250 WORDS)


Outcomes of Patients With ST-Segment Elevation Myocardial Infarction Admitted During COVID-19 Pandemic Lockdown in Germany - Results of a Single Center Prospective Cohort Study.
Objective: Since the outbreak of the COVID-19 pandemic, healthcare professionals reported declining numbers of patients admitted with ST-segment myocardial infarction (STEMI) associated with increased in-hospital morbidity and mortality. However, the effect of lockdown on outcomes of STEMI patients admitted during the COVID-19 crisis has not been prospectively evaluated. Methods: A prospective, observational study on STEMI patients admitted to our tertiary care center during the COVID-19 pandemic was conducted. Outcomes of patients admitted during lockdown were compared to those patients admitted before and after pandemic-related lockdown. Results: A total of 147 patients were enrolled in our study, including 57 patients in the pre-lockdown group (November 1, 2019 to March 20, 2020), 16 patients in the lockdown group (March 21 to April 19, 2020), and 74 patients in the post-lockdown group (April 20 to September 30, 2020). Patients admitted during lockdown had significantly longer time to first medical contact, longer door-to-needle-time, higher serum troponin T levels, worse left ventricular end-diastolic pressure, and higher need for circulatory support. After a median follow-up of 142 days, survival was significantly worse in STEMI patients of the lockdown group (log-rank: p = 0.0035). Conclusions: This is the first prospective study on outcomes of STEMI patients admitted during public lockdown amid the COVID-19 pandemic. Our results suggest that lockdown might deteriorate outcomes of STEMI patients. Public health strategies to constrain spread of COVID-19, such as lockdown, have to be accompanied by distinct public instructions to ensure timely medical care in acute diseases such as STEMI.


Sites of action of procaine at the motor end-plate.
The effects of procaine at pH 7.4 and 9.9 were studied by examining the decay phase of spontaneous miniature end-plate currents (m.e.p.c.s) recorded from toad sartorius muscle fibres. Following exposure to procaine (0.05-0.1 mM) at pH 7.4, the decay of m.e.p.c.s rapidly became biphasic, and could be described as the sum of two exponential components. When the same concentrations of procaine were applied at pH 9.9, the development of biphasic m.e.p.c.s took much longer. Reversal of the effect upon washing out the procaine was much slower at pH 9.9 than at pH 7.4. A rapid change in pH from 7.4 to 9.9 during exposure to a constant concentration of procaine quickly reduced the effect of procaine on m.e.p.c.s. The effect gradually returned after prolonged exposure to procaine at pH 9.9. These results suggest that procaine applied at high pH, where it is predominantly in uncharged form, may be diffusing to a site of action which is not directly accessible from the external surface of the membrane. The voltage-dependence of procaine action was similar whether it was applied at pH 7.4 or 9.9. Intracellular injection of procaine rapidly produced biphasic m.e.p.c.s, whether the extracellular pH was 7.4 or 9.9. The effect of membrane potential on these m.e.p.c.s was similar to that seen for biphasic m.e.p.c.s produced by extracellular application of procaine. The results indicate that procaine can affect end-plate channels when applied to either surface of the muscle membrane, and that the voltage-dependence of procaine action does not arise from the influence of membrane field on the movement of charged procaine molecules into open channels.


[Sutureless anastomoses in gastrointestinal surgery with and without steady magnetic field (experimental study)].
Experimentally in 52 dogs, sutureless gastro-duodenal and ceco-jejunal side-to-side anastomoses were done by means of devices proposed by N.N. Kanshin for connecting void organs, and the features of accretion of the organ walls under the effect of both simple mechanical compression and that with the help of a steady magnetic field were studied. In both variants the process of anastomosis formation completes at 6-7 days after the operation and natural elimination of the compressing elements occurs at 7-8 days. Morphological examinations of sutureless anastomoses formed under a magnetic field or without it revealed no differences between them. The experimental results demonstrated a number of significant advantages of the sutureless methods of tissue connection over the traditional manual intestinal suture.


[Combination of morphine with low-dose naloxone for intravenous patient-controlled analgesia].
To compare the analgesic effect and side effects of morphine for intravenous patient-controlled analgesia (PCA) with or without low-dose naloxone after abdominal surgery. Fifty-nine ASA I - II patients undergoing elective abdominal surgery were randomly divided into two groups: group morphine received postoperative PCA with 0.4 mg/ml morphine (a 1 mg bolus with a 5 min lockout), group naloxone received morphine 0.4 mg/ml with 6 microg/kg naloxone. Blood pressure, heart rate, respiratory rate, and pulse oxygen saturation were monitored. Visual analogue scale (VAS), nausea/vomiting, pruritus, sedation and consumption of morphine were recorded for 24 hours. VAS had no difference between group morphine and group naloxone, but group naloxone had significantly lower VAS for pain at rest or movement (beyond 4-8 h), and the incidence of nausea/vomiting significantly decreased in group naloxone (P < 0.05). No differences existed in pruritus, sedation, respiratory rate, and hemodynamic parameters between these two groups. The 24 hours postoperative morphine consumption was (36.6 +/- 13.5) mg in group naloxone and (43.7 +/- 14.6) mg in group morphine (P < 0.05). For morphine PCA, morphine with 6 microg/kg naloxone is effective in preventing some PCA morphinerelated side effects. Naloxone not only reduces postoperative morphine requirements but also improves the analgesic effect.


Diagnostic accuracy of holotranscobalamin, methylmalonic acid, serum cobalamin, and other indicators of tissue vitamin B₁₂ status in the elderly.
Vitamin B₁₂ deficiency is common among the elderly, and early detection is clinically important. However, clinical signs and symptoms have limited diagnostic accuracy and there is no accepted reference test method. In elderly subjects (n = 700; age range 63-97 years), we investigated the ability of serum cobalamin, holotranscobalamin (holoTC), total homocysteine (tHcy), methylmalonic acid (MMA), serum and erythrocyte folate, and other hematologic variables to discriminate cobalamin deficiency, defined as red blood cell cobalamin <33 pmol/L. Serum holoTC was the best predictor, with area under the ROC curve (95% CI) 0.90 (0.86-0.93), and this was significantly better (P ≤ 0.0002) than the next best predictors; serum cobalamin, 0.80 (0.75-0.85), and MMA, 0.78 (0.72-0.83). For these 3 analytes, we constructed a 3-zone partition of positive and negative zones and a deliberate indeterminate zone between. The boundaries were values of each test that resulted in a posttest probability of deficiency of 60% and a posttest probability of no deficiency of 98%. The proportion of indeterminate observations for holoTC, cobalamin, and MMA was 14%, 45%, and 50%, respectively. Within the holoTC indeterminate zone (defined as 20-30 pmol/L), discriminant analysis selected only erythrocyte folate, which correctly allocated 65% (58/89) of the observations. Renal dysfunction compromised the diagnostic accuracy of MMA but not holoTC or serum cobalamin. This study supports the use of holoTC as the first-line diagnostic procedure for vitamin B₁₂ status.


Glycinergic pathways in the goldfish retina.
Autoradiographic localization of high affinity [3H]glycine uptake in the retina of the goldfish has been used to study some anatomical and physiological properties of potentially glycinergic neurons. There are two classes of retinal cells exhibiting high affinity glycine uptake: Aa amacrine cells and I2 interplexiform cells. Aa amacrine cells constitute about 20% of the somas in the amacrine cell layer and send their dendrites to the middle of the inner plexiform layer. There they are both pre- and postsynaptic primarily to other amacrine cells. Photic modulation of glycine uptake indicates that they are probably red-hyperpolarizing/green-depolarizing neurons. I2 interplexiform cells are a newly discovered type of interplexiform cell; in the outer plexiform layer, they receive direct synaptic input from the somas of red-dominated GABAergic H1 horizontal cells and are apparently presynaptic to dendrites of unidentified types of horizontal cells. The connections of I2 interplexiform cells have not been successfully characterized in the inner plexiform layer. These findings extend our knowledge of neurochemically specific pathways in the cyprinid retina and indicate that glycine, like GABA, is a neurotransmitter primarily involved with circuits coding "red" information.


Cardiac mucormycosis: a case report.
Mucormycosis is an invasive fungal infection (IFI) most commonly seen in immunocompromised patients. Diabetic ketoacidosis, haematopoietic transplantation, iron overload states, and deferoxamine therapy are considered to be some of the classical risk factors. While cutaneous and rhino-sinusoidal forms may be seen in immunocompetent (IC) individuals, cardiac and mediastinal involvement is rare. In this report, we describe a young patient without predisposing factors who presented as mediastinal mucormycosis with extensive cardiac involvement. A 19-year-old male presented with complaints of dry cough and dyspnoea on exertion over the last 4 months. Echocardiography showed diffuse infiltration of both atria along with multiple pedunculated freely mobile masses. A computed tomography chest was done to further delineate the true extent of the disease and revealed diffuse infiltration of the mediastinum, bilateral atria and interatrial septum, pulmonary veins, and superior vena cava. A fine needle aspiration cytology from a mediastinal mass revealed broad aseptate fungal hyphae with right angled branching consistent with Mucor. Extensive evaluation could not find any predisposing factors. The patient was started on Amphotericin B and surgical debridement was contemplated. However, owing to the diffuse infiltration around the heart and mediastinal vasculature, debridement could not be performed and the patient eventually succumbed to the illness. Mediastinal mucormycosis though rare in IC patients, is a rapidly progressive condition with a high fatality. A high index of suspicion needs to be maintained in individuals presenting with infiltrative disorders of the mediastinum for early diagnosis and prompt treatment.


TCF7L2, dietary carbohydrate, and risk of type 2 diabetes in US women.
TCF7L2 is the strongest type 2 diabetes (T2D) locus identified to date, and evidence suggests it plays an important role in insulin synthesis, processing, and secretion. Dietary factors that increase the insulin demand might enhance the risk of T2D associated with TCF7L2 variants. The objective was to determine whether the risk of T2D associated with TCF7L2 is modified by the glycemic load (GL), glycemic index (GI), cereal fiber content, and total carbohydrate content of the diet. T2D cases (n = 1140) and controls (n = 1915) from the Nurses' Health Study were genotyped for TCF7L2 (rs12255372). Dietary intake was assessed with a semiquantitative food-frequency questionnaire. Significant differences in odds ratios (ORs) of T2D associated with the TCF7L2 genotype between high and low strata of GL (P = 0.03) and GI (P = 0.05) were suggested. Compared with the GG genotype, multivariate-adjusted ORs (95% CI) of T2D associated with the TT genotype were 2.71 (1.64, 4.46) and 2.69 (1.64, 4.43) among individuals in the highest tertile of GL and GI, respectively. Corresponding ORs (95% CIs) among individuals in the lowest tertiles of GL and GI were 1.66 (0.95, 2.88) and 1.82 (1.11, 3.01). The risk of T2D associated with the TCF7L2 single nucleotide polymorphism did not significantly differ by cereal fiber or carbohydrate intake. Carbohydrate quality and quantity modified risk of T2D associated with TCF7L2, which suggests that changes in risk attributable to the TCF7L2 variant are magnified under conditions of increased insulin demand.


Econazole-polycarbophil, a new delivery system for topical therapy: microbiological and clinical results on vaginal candidiasis.
The aim of this study was to demonstrate that the addition of a bioadhesive polymer to econazole, which increases the duration of the active drug at the site of infection, leads to a greater frequency of negative culture after treatment and probably reduces the recurrence rate of vaginal candidiasis.180 women with vaginal candidiasis were treated with 150 mg vaginal ovules econazole nitrate with (group A) or without (group B) polycarbophil. After 3 days of treatment the negative culture of Candida albicans reached 98.6% in group A and 84.8% in B group, while the overall persistence (C. albicans, C. glabrata, C. krusei, and C. parapsilosis) was 5.6% and 30%, respectively. During a 60-day follow-up, only one case out of 85 (1.2%) in group A reported recurrence while in group B there were 6 out of 63 (9.5%) recurrences. We conclude that, since the women were treated with the same amount of econazole, the better clinical and microbiological results can be attributed to polycarbophil, as confirmed by a significant reduction of recurrences.


Strontium modified calcium phosphate cements - approaches towards targeted stimulation of bone turnover.
Making use of the potential of calcium phosphates to host a variety of ions in their crystal lattice, ion substitution of calcium phosphate bone cements has become the subject of intense investigations in the last few years, since this approach allows one to stabilize a bone defect and to locally deliver therapeutic ions into a specific defect site at the same time. In this respect significant attention has been given to strontium ions (Sr2+) lately. Strontium possesses the unique potential to both stimulate new bone formation and inhibit cell-driven bone resorption and thus has been used successfully in systemic osteoporosis therapy. Strontium doping of calcium phosphate bone cements might allow making use of this dual effect to promote local bone defect healing. The goal of this review is to provide an overview of different routes that have been employed to obtain strontium-containing calcium phosphate bone cements and describe their material characteristics as well as their biological properties based on cell culture and animal studies.


Nutritional status of urban and rural primary school pupils in Lagos State, Nigeria.
This study was carried out in Lagos State in Ikorodu LGA (rural), a largely agrarian area producing raw materials for agro-allied industries and Ikeja LGA (urban), a highly-developed metropolis and commercial centre. Hitherto, studies showed that undernutrition had been the main form of malnutrition among Nigerian children. However with influences from urbanization, westernization and globalization in cosmopolitan Lagos, it is imperative to find out if the global epidemic of overweight and obesity has caught on. To compare the nutritional status of primary school pupils in urban and rural LGAs in Lagos State, Nigeria. This was a cross , sectional comparative study. A multistage sampling method was adopted. Data were collected from 529 pupils with an interviewer administered questionnaire. Their weight and height were recorded in pre-designed proforma and converted to nutritional indices. The prevalence rates of underweight, stunting and wasting in the rural area were 49.6%, 50.8% and 24.2% respectively; while it was 15.1%, 16.6% and 13.6% in the urban area respectively. There was overnutrition in the urban area: 15.1% were overweight and 13.2% were obese. The mean nutritional indices were significantly lower among the rural pupils than urban pupils (p<0.001) in each case. Undernutrition is still a major health problem among school children in Lagos State, Nigeria, more so in the rural area. There is an emergence of overweight and obesity in the urban area. A comprehensive programme to improve nutritional status is recommended as part of a well funded school health programme.


Adrenergic regulation of avian pancreatic polypeptide secretion in vitro.
Adrenergic regulation of avian pancreatic polypeptide (APP) secretion from perifused microfragments of chicken pancreas was investigated with catecholamines and adrenoreceptor-specific analogues added to avian Krebs-Ringer perifusion medium containing 11.1 mM glucose. APP release was stimulated by the alpha-adrenoreceptor agonist phenylephrine (threshold 5 microM; maximum effective concn, 50 microM) and the beta-adrenoreceptor agonist isoproterenol (threshold 5 microM; maximum effective concn, 50 microM). Epinephrine (0.164 microM) and 50 microM norepinephrine also stimulated APP release. The results obtained suggest that APP secretion from chicken pancreas is regulated by dual, complementary alpha- and beta-adrenoreceptor mechanisms.


Modulation of the plasminogen activator cascade during enhanced epidermal proliferation in vivo.
Many lines of evidence support an involvement of urokinase plasminogen activator (uPA) and its type 1 inhibitor (PAI-1) in the migration of a variety of cells, including normal keratinocytes and carcinoma lines. In the present study, uPA expression was found to be a characteristic not just of migratory but also of proliferative keratinocytes. A variety of naturally occurring and experimentally induced epidermal hyperproliferative conditions were examined in mice, including fetal and neonatal epidermis, tape-stripped epidermis, and epidermis from which the hairs had been gently plucked. In all cases, epidermal hyperproliferation was accompanied by elevated levels of uPA mRNA (as measured by in situ hybridization) and activity (as measured by zymography). uPA mRNA was predominantly localized in the basal and immediately suprabasal cells, which constitute the proliferative population. To determine whether a PAI was concomitantly elevated, in situ hybridization for PAI-1 and PAI-2 was performed. PAI-2 but not PAI-1 mRNA was detected in fetal and neonatal epidermis, localized in the spinous layers. Although mRNAs for both inhibitors were induced by tape-stripping or hair-plucking, their distribution was more focal and more transient than that of uPA mRNA. These findings show that uPA, but not its usual inhibitors, is consistently elevated in the proliferative population of keratinocytes in a diverse range of hyperproliferative states. Two hypotheses are suggested by these data: (a) uPA may play a regulatory role in the activation of epidermal proliferation; or (b) uPA may be involved in the vertical migration of keratinocytes that must accompany increased cell proliferation.


ROS-Induced mtDNA Release: The Emerging Messenger for Communication between Neurons and Innate Immune Cells during Neurodegenerative Disorder Progression.
One of the most striking hallmarks shared by various neurodegenerative diseases, including Parkinson's disease, Alzheimer's disease and amyotrophic lateral sclerosis, is microglia-mediated and astrocyte-mediated neuroinflammation. Although inhibitions of both harmful proteins and aggregation are major treatments for neurodegenerative diseases, whether the phenomenon of non-normal protein or peptide aggregation is causally related to neuronal loss and synaptic damage is still controversial. Currently, excessive production of reactive oxygen species (ROS), which induces mitochondrial dysfunction in neurons that may play a key role in the regulation of immune cells, is proposed as a regulator in neurological disorders. In this review, we propose that mitochondrial DNA (mtDNA) release due to ROS may act on microglia and astrocytes adjacent to neurons to induce inflammation through activation of innate immune responses (such as cGAS/STING). Elucidating the relationship between mtDNA and the formation of a pro-inflammatory microenvironment could contribute to a better understanding of the mechanism of crosstalk between neuronal and peripheral immune cells and lead to the development of novel therapeutic approaches to neurodegenerative diseases.


Enrichment and efficient screening of ES cells containing a targeted mutation: the use of DT-A gene with the polyadenylation signal as a negative selection maker.
Gene targeting in embryonic stem (ES) cells via homologous recombination can occur at very low frequency. In order to enrich homologous recombinants before screening, a negative selection marker, such as thymidine kinase (TK) and diphtheria toxin A fragment (DT-A), has been commonly used. In this study, we developed a negative selection marker using DT-A gene with polyadenylation signal and it was designated DT-ApA. To determine the difference in targeting efficiency of the negative selections, we constructed three different targeting vectors for each negative selection (first, TK at the 3' end, second, TK at both the 5' and 3' ends < 2 X TK >, and third, DT-ApA at the 5' end of the homologous sequences). Gene targeting experiments using these constructs clearly showed that negative selection using DT-ApA was more efficient than that using TK for homologous recombination and that negative selection using DT-ApA was as efficient as that using 2 X TK. Considering the fact that the use of DT-ApA is more convenient for construction of targeting vectors than that of 2 X TK, DT-ApA is an efficient negative selection marker. In addition, we examined long and accurate PCR (LA-PCR) for screening gene targeted clones. The use of LA-PCR with genomic DNAs from ES cell clones facilitated simple detection of homologous recombinants, suggesting that the screening with LA-PCR is compatible with the use of longer homologous sequences of both arms in vector design. Our results indicate that the use of DT-ApA for negative selection together with the application of LA-PCR for screening ensures efficient and time-saving screening for homologous recombinants.


Evaluation of integrity of gloves used in a flow cytometry laboratory.
This study was conducted to evaluate the effect of normal use on latex glove integrity in a flow cytometry laboratory. The gloves were tested using the 1,000 microL water-tight test and met industrial standards (less than 4% leakage) before, but not after use. More durable gloves, or more frequent changes of gloves, may be needed to ensure adequate barrier protection for laboratory workers during routine procedures.


A molecular phylogenetic analysis of strombid gastropod morphological diversity.
The shells of strombid gastropods show a wide variety of forms, ranging from small and fusiform to large and elaborately ornamented with a strongly flared outer lip. Here, we present the first species-level molecular phylogeny for strombids and use the resulting phylogenetic framework to explore relationships between species richness and morphological diversity. We use portions of one nuclear (325 bp of histone H3) and one mitochondrial (640 bp of cytochrome oxidase I, COI) gene to infer relationships within the two most species-rich genera in the Strombidae: Strombus and Lambis. We include 32 species of Strombus, representing 10 of 11 extant subgenera, and 3 of the 9 species of Lambis, representing 2 of 3 extant subgenera. Maximum likelihood and Bayesian analyses of COI and of H3 and COI combined suggest Lambis is nested within a paraphyletic Strombus. Eastern Pacific and western Atlantic species of Strombus form a relatively recent monophyletic radiation within an older, paraphyletic Indo-West Pacific grade. Morphological diversity of subclades scales positively with species richness but does not show evidence of strong phylogenetic constraints.


Additive neurocognitive deficits in adults with attention-deficit/hyperactivity disorder and depressive symptoms.
The purpose of this study was to examine the possible additive neurocognitive deficits in adults with both attention-deficit/hyperactivity disorder (ADHD) and serious depressive symptoms. Participants were 54 university students who completed a psycho-educational assessment. Three groups were examined: a group with comorbid ADHD and elevated depressive symptoms (ADHD + DEP; N = 18); a group with ADHD only (N = 18); and a group with elevated depressive symptoms only (DEP; N = 18). Group differences were examined on a battery of neurocognitive tests. The ADHD + DEP group performed significantly worse than the other groups on processing speed tasks and delayed recall of conceptual verbal information and significantly worse than the ADHD group on shifting tasks. Depressive symptom severity was significantly correlated with processing speed, verbal memory performance, and shifting in the ADHD and ADHD + DEP groups. Results suggest that the co-occurrence of ADHD and depressive symptoms in adults is associated with additional neurocognitive impairment.


Loss of the tight junction protein claudin-7 correlates with histological grade in both ductal carcinoma in situ and invasive ductal carcinoma of the breast.
Claudins are transmembrane proteins that seal tight junctions, and are critical for maintaining cell-to-cell adhesion in epithelial cell sheets. However, their role in cancer progression remains largely unexplored. Here, we report that Claudin-7 (CLDN-7) expression is lower in invasive ductal carcinomas (IDC) of the breast than in normal breast epithelium, as determined by both RT-PCR (9/10) and Western analysis (6/8). Immunohistochemical (IHC) analysis of ductal carcinoma in situ (DCIS) and IDC showed that the loss of CLDN-7 expression correlated with histological grade in both DCIS (P<0.001, n=38) and IDC (P=0.014, n=31), occurring predominantly in high-grade (Nuclear and Elston grade 3) lesions. Tissue array analysis of 355 IDC cases further confirmed the inverse correlation between CLDN-7 expression and histological grade (P=0.03). This pattern of expression is consistent with the biological function of CLDN-7, as greater discohesion is typically observed in high-grade lesions. In line with this observation, by IHC analysis, CLDN-7 expression was lost in the vast majority (13/17) of cases of lobular carcinoma in situ, which is defined by cellular discohesion. In fact, inducing disassociation of MCF-7 and T47D cells in culture by treating with HGF/scatter factor resulted in a loss of CLDN-7 expression within 24 h. Silencing of CLDN-7 expression correlated with promoter hypermethylation as determined by methylation-specific PCR (MSP) and nucleotide sequencing in breast cancer cell lines (3/3), but not in IDCs (0/5). In summary, these studies provide insight into the potential role of CLDN-7 in the progression and ability of breast cancer cells to disseminate.


Indirect costs associated with early exit from work attributable to rheumatic diseases.
Rheumatic diseases (RD) cause physical disability that may lead to early exit from work, generating indirect costs to society. We aimed to measure these costs in a population approaching the statutory retirement age. The analysis was based on the prevalence of self-reported RD using a bottom-up approach. Health and sociodemographic data were retrieved from the fourth National Health Survey (INS), for all people between 50 and 64 years of age (3762 men and 4241 women), whereas an official national database was used to estimate productivity values by gender, age group and region, using the human capital approach. The effects of RD on the likelihood of early exit from paid employment and the attributable fractions estimates were obtained at the individual level by logistic regression. At the time of the survey, 37.2% of the population aged 50-64 years self-reported at least one RD. Among these, 52.6% were not employed, compared with 40.7% of those without RD (P < 0.001). The annual indirect costs following premature exit from work attributable to RD were €650 million (€892 per RD patient). Early retirement amounted to €367 million, whereas early retirement and unemployment totalized €385 million (€504 and €528 per RD patient, respectively). Females are responsible for about 60% of these costs; however, males contribute with higher individual productivity losses. Early exit from work attributable to RD amounts to approximately 0.4% of the national GDP. The public health concern and the economic impact highlight the need to prioritize investments in health and social protection policies targeting patients with rheumatic conditions.


Drug repurposing in pediatrics and pediatric hematology oncology.
Drug 'repurposing', that is, using old drugs for new indications, has been proposed as a more efficient strategy for drug development than the current standard of beginning with novel agents. In this review, we explore the scope of drug repurposing in pediatric hematology oncology and in pediatrics in general. Drugs commonly used in children were identified using the Harriet Lane Handbook (HLH) and searched in PubMed for different uses. Additional drugs were identified by searching PubMed and Google.com for 'drug repurposing' or 'drug repositioning'. Almost 10% of drugs with primary uses in pediatrics have been repurposed in pediatric hematology oncology or pediatrics. The observant clinician, pharmacologist and translational bioinformatician, as well as structural targeting, will have a role in discovering new repurposing opportunities.


Quality of life after proctocolectomy. A comparison of Brooke ileostomy, Kock pouch, and ileal pouch-anal anastomosis.
Recent work has shown that patients undergoing proctocolectomy and ileal pouch-anal anastomosis experience a better quality of life than those undergoing proctocolectomy and Brooke ileostomy. To assess whether the improvement with the pouch is due to the absence of a stoma or the preservation of fecal continence, functional and performance activities were assessed in 406 patients with Brooke ileostomies (stoma present, incontinent), 313 with Kock pouches (stoma present, continent), and 298 with ileal pouch-anal anastomoses (stoma absent, continent). All patients underwent proctocolectomy for ulcerative colitis or familial adenomatous polyposis. After adjusting for age, sex, diagnosis, attitude towards the operation, dietary satisfaction, and ability to work, the likelihood of an impact on performance was estimated for the three types of operation using logistic regression analysis. Patients with ileal pouch-anal anastomoses had fewer restrictions in sports and sexual activities than those with Kock pouches (P less than 0.05), whereas those with Kock pouches in turn had fewer restrictions in these activities but more restrictions in travel than those with Brooke ileostomies (P less than 0.05). In contrast, performance in the categories of social life, recreation, work, and family was similar between groups. It is concluded that both the presence of a stoma and fecal incontinence impair the quality of life after proctocolectomy. Ileal pouch-anal anastomosis, which avoids both stoma and incontinence, offers the best quality of life between the three operations studied.


Ovarian adenocarcinomas express fms-complementary transcripts and fms antigen, often with coexpression of CSF-1.
In earlier studies of oncogene expression in ovarian and endometrial neoplasms, the authors reported that high tumor levels of fms-complementary transcripts correlate with high histologic grade and advanced clinical stage presentations. In this communication, they pursue these initial clinicopathologic investigations to demonstrate by in situ hybridization and immunohistochemistry that malignant epithelial cells of 14 of 14 invasive adenocarcinomas of the ovary express fms-complementary transcripts. By Northern blotting and by reverse transcription, followed by polymerase chain reaction amplification, the authors also were able to demonstrate fms transcript expression in several ovarian and endometrial carcinoma-derived cell lines. Because about half (6/14) of the invasive adenocarcinoma specimens were shown to coexpress fms and colony-stimulating factor 1, the authors propose that the expression of this lymphohematopoietic cytokine and its receptor by ovarian adenocarcinomas could contribute to their proliferative and invasive characteristics in vivo.


eHealth Literacy: Patient Engagement in Identifying Strategies to Encourage Use of Patient Portals Among Older Adults.
Innovations in chronic disease management are growing rapidly as advancements in technology broaden the scope of tools. Older adults are less likely to be willing or able to use patient portals or smartphone apps for health-related tasks. The authors conducted a cross-sectional survey of older adults (ages ≥50) with hypertension or diabetes to examine relationships between portal usage, interest in health-tracking tools, and eHealth literacy, and to solicit practical solutions to encourage technology adoption. Among 247 patients surveyed in a large integrated delivery health system between August 2015 and January 2016, eHealth literacy was positively associated with portal usage (OR [95% CI]: 1.3 [1.2-1.5]) and interest in health-tracking tools (1.2 [1.1-1.3]). Portal users compared to nonusers (N = 137 vs.110) had higher rates of interest in using websites/smartphone apps to track blood pressure (55% vs. 36%), weight (53% vs. 35%), exercise (53% vs. 32%), or medication (46% vs 33%, all P < 0.05). Portal users noted cumbersome processes for accessing portals and variations in provider availability for online scheduling and response times to messages. Portal nonusers expressed concerns about data security, lack of personalization, and limited perceived value of using portals. Both groups noted the importance of computer literacy and technical support. Patient stakeholders recommended marketing initiatives that capture patient stories demonstrating real-life applications of what patients can do with digital technology, how to use it, and why it may be useful. Health systems also must screen for eHealth literacy, provide training, promote proxy users, and institute quality assurance that ensures patients' experiences will not vary across the system.


Plant-parasitic Nematode Communities and Their Associations with Soil Factors in Organically Farmed Fields in Minnesota.
A survey was conducted to determine the assemblage and abundance of plant-parasitic nematodes and their associations with soil factors in organically farmed fields in Minnesota. A total of 31 soil samples were collected from southeast (SE), 26 samples from southwest (SW), 28 from west-central (WC), and 23 from northwest (NW) Minnesota. The assemblage and abundance of plant-parasitic nematodes varied among the four regions. The soybean cyst nematode, Heterodera glycines, the most destructive pathogen of soybean, was detected in 45.2, 88.5, 10.7, and 0% of organically farmed fields with relative prominence (RP) values of 10.3, 26.5, 0.6, and 0 in the SE, SW, WC, and NW regions, respectively. Across the four regions, other common genera of plant-parasitic nematodes were Helicotylenchus (42.6, RP value, same below), Pratylenchus (26.9), Tylenchorhynchus and related genera (9.4), Xiphinema (5.6), and Paratylenchus (5.3). Aphelenchoides, Meloidogyne, Hoplolaimus, Mesocriconema, and Trichodorus were also detected at low frequencies and/or low population densities. The similarity index of plant-parasitic nematodes between two regions ranged from 0.44 to 0.71 and the similarity increased with decreasing distance between regions. The densities of most plant-parasitic nematodes did not correlate with measured soil factors (organic matter, pH, texture). However, the densities of Pratylenchus correlated negatively with % sand, and Xiphinema was correlated negatively with soil pH.