Salivary sialic acid levels and dental health in children with congenital heart disease.
Congenital cardiac disease is one of the most common developmental anomalies in children. Higher caries prevalence and gingivitis compared to healthy children have been shown in children with congenital heart disease which has a significant implication in the medical care of these patients associated with bacteraemia and endocarditis. Sialic acids being terminal sugar components and marker of chronic inflammatory response are found to be present at higher levels in children with poor oral health status. So the present study aimed to evaluate the oral health status and salivary sialic acid levels among the children with congenital heart disease and normal healthy siblings. A total of 71 children with heart diseases aged 6-10 years attending various heart institutes were examined for oral hygiene status, gingival status and dental caries status by using modified WHO oral assessment form. A case-matched control group of 50 children were also examined. Salivary flow rate, pH and sialic acid levels were measured after saliva collection. The results were subjected to Unpaired t test and Pearson's Correlation Coefficient Test. The salivary pH, salivary flow rate, dental caries status, oral hygiene and the gingival status were significantly compromised with a positive correlation with the sialic acid levels in saliva, leading to increased treatment needs in the study group. Therefore the amount of sialic acid in the saliva can be a useful index of the severity of oral disease.


Hepatitis B infection in patients with acute liver failure in the United States.
Occult hepatitis B virus (HBV) infection has been reported in 30% to 50% of patients with acute liver failure (ALF) in small case series. The aim of this study was to determine the prevalence of occult HBV infection in a large series of ALF patients in the United States and the prevalence of precore and core promoter variants in patients with ALF caused by hepatitis B. Sera from patients in the US ALF study and liver, when available, were tested using nested polymerase chain reaction (PCR) with primers in the HBV S and precore regions. PCR-positive samples were sequenced. Sera and/or liver from 139 patients (39 males, 100 females; mean age, 42 years) enrolled between January 1998 and December 1999 were studied. Twelve patients were diagnosed with hepatitis B, 1 with hepatitis B+C+D coinfection, and 22 had indeterminate etiology. HBV DNA was detected in the sera of 9 (6%) patients; all 9 had ALF caused by hepatitis B. HBV genotypes A, B, C, and D were present in 4, 3, 1, and 1 patients, respectively. Seven of these 9 patients had precore and/or core promoter variants. Liver from 19 patients were examined. HBV DNA was detected in the liver of 3 patients with ALF caused by hepatitis B, but in none of the remaining 16 patients with non-B ALF. Contrary to earlier reports, occult HBV infection was not present in this large series of ALF patients in the United States. HBV precore and/or core promoter variants were common among US patients with ALF caused by hepatitis B.


Targeting Cell Necroptosis and Apoptosis Induced by Shikonin via Receptor Interacting Protein Kinases in Estrogen Receptor Positive Breast Cancer Cell Line, MCF-7.
Recognition of a new therapeutic agent may activate an alternative programmed cell death for the treatment of breast cancer. Here, it has been tried to evaluate the effects of Shikonin, a naphthoquinone derivative of Lithospermum erythrorhizon, on the induction of necroptosis and apoptosis mediated by RIPK1-RIPK3 in the ER+ breast cancer cell line, MCF-7. In the current study, cell death modalities, cell cycle patterns, RIPK1 and RIPK3 expressions, caspase-3 and caspase-8 activities, reactive oxygen species and mitochondrial membrane potential have been evaluated in the Shikonin-treated MCF-7 cells. Necroptosis and apoptosis have been occurred by Shikonin, with a significant increase in RIPK1 and RIPK3 expressions, although necroptosis was the major rout in MCF-7 cells. Shikonin significantly increased the percentage of the cells in sub-G1 and also those in the later stages of cell cycle, which represents an increase in necroptosis and apoptosis. Under caspase inhibition by Z-VAD-FMK, Shikonin has stimulated necroptosis, which could be arrested by Nec-1. An increase in ROS levels and a decrease in the mitochondrial membrane potential have also been observed. On the basis of present findings, Shikonin has been suggested as a good candidate for the induction of cell death in ER+ breast cancer, although further investigations, experimental and clinical, are required.


Effects of zopiclone as compared to flurazepam on sleep in women over 40 years of age.
Zopiclone is a new nonbenzodiazepine hypnotic, the recommended dose of which is 7.5 mg. It was compared to flurazepam 30 mg in a sleep laboratory study. 12 women aged 40-60 years, with subjective sleep disorders for at least 2 months, were treated for 13 nights with each drug in a crossover double-blind design. Each active drug period was preceded by 5 days of placebo washout and followed by 35 days of withdrawal (no treatment). A baseline period of 4 nights (without treatment) was obtained before the study itself. The subjects slept in the sleep laboratory during the baseline period, during the last 3 nights on placebo, during the last 3 nights on active drug, and from the 11th to the 13th night after active drug discontinuation. Polygraphic recordings were obtained during each of these nights. The criteria of Kales and Rechtschaffen were used to score the tracings. Both drugs were shown to be effective when compared to placebo. Comparisons between the two periods with active treatment did not show significant differences for sleep onset latency, total sleep time, sleep efficiency index, stage 0, percent REM, number of REM periods, and REM latency. There was a trend towards an increase with zopiclone of the number of awakenings, percent of stage 3, percent of deep sleep, and an increase with flurazepam of the percent of light sleep. A significant difference was only found for percent of stage 2 (increased with flurazepam).


Prevalence of risk factors for HIV infection among Mexican migrants and immigrants: probability survey in the North border of Mexico.
To estimate the prevalence of risk factors for HIV infection among Mexican migrants and immigrants (MMIs) in different geographic contexts, including the sending communities in Mexico, the receiving communities in the United States (US), and the Mexican North border region. We conducted a probability survey among MMIs traveling through key border crossing sites in the Tijuana (Baja California, Mexico)-San Diego (California, US) border region (N= 1,429). The survey revealed substantial rates of reported sexually transmitted infections, needle-sharing and sexual risk practices in all migration contexts. The estimated levels of HIV risk call for further binational research and preventive interventions in all key geographic contexts of the migration experience to identify and tackle the different personal, environmental, and structural determinants of HIV risk in each of these contexts.


Detection of Fumonisins in Fusarium Cultures, Corn, and Corn Products by Polyclonal Antibody-Based ELISA: Relation to Fumonisin B1 Detection by Liquid Chromatography.
A specific sheep polyclonal antiserum was applied to the competitive direct enzyme-linked immunosorbent assay (CD-ELISA) of fumonisins in Fusarium cultures, fresh corn, animal feed, and human foods, and the results were related to fumonisin B1 (FB1) levels determined by liquid chromatography (LC). The limit of detection of FB1 for the CD-ELISA was 0.1 ng/ml. Cross reactivity (defined as [ng/ml FB1 required for 50% inhibition]/[ng/ml of analogue required for 50% inhibition] × 100) for the antiserum in the ELISA was 100, 24, and 30%, for FB1 FB2, and FB3, respectively. FB1 was detected in F. moniliforme cultures grown on corn (≤760 μg/g) but not in those for F. graminearum . Fumonisin estimates were 2.8-fold higher by CD-ELISA than FB1 estimates by LC. Upon using the antiserum in a commercial CD-ELISA format (Veratox), mean recovery of FB1 from spiked corn (0.1 to 3 μg/g)was 85.2 ± 25.3%, whereas the mean recovery by LC was 74.1 ± 6.2%. Analyses of 43 fresh corn, 28 animal feed, and 14 human food samples revealed that estimates of total fumonisins by the commercial ELISA were 2.9-, 2.2-, and 1.3-fold higher than FB1 values determined by LC. For each sample type, the ELISA significantly correlated with LC (r > 0.9; P < 0.05). Taken together, the results suggest that the polyclonal antibody-based CD-ELISA could be a reliable first-tier screening procedure for fumonisins in corn, animal feed, and human food when used in conjunction with LC confirmation. The differences between the two methods suggests the further presence of FB2 and FB3 or structurally related compounds.


It takes a village: A multi-brain approach to studying multigenerational family communication.
Grandparents play a critical role in child rearing across the globe. Yet, there is a shortage of neurobiological research examining the relationship between grandparents and their grandchildren. We employ multi-brain neurocomputational models to simulate how changes in neurophysiological processes in both development and healthy aging affect multigenerational inter-brain coupling - a neural marker that has been linked to a range of socio-emotional and cognitive outcomes. The simulations suggest that grandparent-child interactions may be paired with higher inter-brain coupling than parent-child interactions, raising the possibility that the former may be more advantageous under certain conditions. Critically, this enhancement of inter-brain coupling for grandparent-child interactions is more pronounced in tri-generational interactions that also include a parent, which may speak to findings that grandparent involvement in childrearing is most beneficial if the parent is also an active household member. Together, these findings underscore that a better understanding of the neurobiological basis of cross-generational interactions is vital, and that such knowledge can be helpful in guiding interventions that consider the whole family. We advocate for a community neuroscience approach in developmental social neuroscience to capture the diversity of child-caregiver relationships in real-world settings.


Estimating Local Costs Associated With Clostridium difficile Infection Using Machine Learning and Electronic Medical Records.
BACKGROUND Reported per-patient costs of Clostridium difficile infection (CDI) vary by 2 orders of magnitude among different hospitals, implying that infection control officers need precise, local analyses to guide rational decision making between interventions. OBJECTIVE We sought to comprehensively estimate changes in length of stay (LOS) attributable to CDI at a single urban tertiary-care facility using only data automatically extractable from the electronic medical record (EMR). METHODS We performed a retrospective cohort study of 171,938 visits spanning a 7-year period. In total, 23,968 variables were extracted from EMR data recorded within 24 hours of admission to train elastic-net regularized logistic regression models for propensity score matching. To address time-dependent bias (reverse causation), we separately stratified comparisons by time of infection, and we fit multistate models. RESULTS The estimated difference in median LOS for propensity-matched cohorts varied from 3.1 days (95% CI, 2.2-3.9) to 10.1 days (95% CI, 7.3-12.2) depending on the case definition; however, dependency of the estimate on time to infection was observed. Stratification by time to first positive toxin assay, excluding probable community-acquired infections, showed a minimum excess LOS of 3.1 days (95% CI, 1.7-4.4). Under the same case definition, the multistate model averaged an excess LOS of 3.3 days (95% CI, 2.6-4.0). CONCLUSIONS In this study, 2 independent time-to-infection adjusted methods converged on similar excess LOS estimates. Changes in LOS can be extrapolated to marginal dollar costs by multiplying by average costs of an inpatient day. Infection control officers can leverage automatically extractable EMR data to estimate costs of CDI at their own institutions. Infect Control Hosp Epidemiol. 2017;38:1478-1486.


Src family kinases regulate p38 MAPK-mediated IL-6 production in Kupffer cells following hypoxia.
Tissue hypoxia is a common sequel of trauma-hemorrhage but can occur even without blood loss under hypoxic conditions. Although hypoxia is known to upregulate Kupffer cells (KC) to release cytokines, the precise mechanism of release remains unknown. We hypothesized that Src family kinases play a role in mediating KC mitogen-activated protein kinase (MAPK) signaling and their cytokine production after hypoxia. Male C3H/HeN mice received either Src inhibitor PP1 (1.5 mg/kg body wt) or vehicle 1 h before hypoxia. KCs were isolated 1 h after hypoxia, lysed, and immunoblotted with antibodies to Src, p38, ERK1/2, or JNK proteins. In addition, KCs were cultured to measure interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) production. Hypoxia produced a significant increase in KC Src and MAPK (p38, ERK, JNK) activity compared with normoxic controls. This was associated with an increase in IL-6 and MCP-1 production. Treatment with PP1 abolished the increase in KC Src activation as well as p38 activity. However, PP1 did not prevent the increase in KC ERK1/2 or JNK phosphorylation. Furthermore, administration of PP1 prevented the hypoxia-induced increase in IL-6 but not MCP-1 release by KC. Additional in vitro results suggest that p38 but not ERK1/2 or JNK are critical for KC IL-6 production. In contrast, the production of MCP-1 by KC was found to be independent of MAPK. Thus hypoxia increases KC IL-6 production by p38 MAPK activation via Src-dependent pathway. Src kinases may therefore be a novel therapeutic target for preventing immune dysfunction following low-flow conditions in trauma patients.


[Hemostasis in patients with ischemic stroke].
Modern informative techniques were employed to explore the coagulation, fibrinolytic and aggregation platelet systems of hemostasis in 117 patients in the most acute , acute, early, late restorative and residual periods of ischemic brain stroke. Disorders of the hemostatic system in the form of hyperaggregation, hypercoagulation, depression of fibrinolysis, and an increase of blood viscosity were revealed as were some specific features marking different period of brain stroke. Mono- and multi-systemic disorders of hemocoagulation and three types of coagulopathies were distinguished. Emphasis is laid on the necessity and feasibility of a goal-oriented selective correction of the pathological conditions of hemostasis at different periods of brain stroke by means of administering appropriate drugs.


Establishment of oral squamous cell carcinoma cell line and magnetic bead-based isolation and characterization of its CD90/CD44 subpopulations.
In this study, we describe the establishment of the human papillomavirus 18-positive, stage II, grade 1, T2N0M0 head and neck tumor primary cell line derived from oral squamous cell carcinoma of a non-smoking patient by using two different protocols. Furthermore, a preparation of subpopulations derived from this primary cell line according to the cluster of differentiation molecules CD44/CD90 status using magnetic bead-based separation and their characterization was performed. Impedance-based real-time cell analysis, enzyme-linked immunsorbant assay (ELISA), wound-healing assay, flow-cytometry, gene expression analysis, and MTT assay were used to characterize these four subpopulations (CD44+/CD90-, CD44-/CD90-, CD44+/CD90+, CD44-/CD90-). We optimised methodics for establishement of primary cell lines derived from oral squamous cell carcinoma tissue samples and subsequent separation of mesenchymal (CD90+) and epithelial (CD90-) types of tumorous cells. Primary cell line prepared by using trypsin proteolysis was more viable than the one prepared by using collagenase. According to our results, CD90 separation is a necessary step in preparation of permanent tumor-tissue derived cell lines. Based on the wound-healing assay, CD44+ cells exhibited stronger migratory capacity than CD44- subpopulations. CD44+ subpopulations had also significantly higher expression of BIRC5 and SOX2, lower expression of FLT1 and IL6, and higher levels of basal autophagy compared to CD44- subpopulations. Furthermore, co-cultivation experiments revealed that CD44-/CD90+ cells supported growth of epithelial tumor cells (CD44+/CD90-). On the contrary, factors released by CD44+/CD90+ type of cells seem to have rather inhibiting effect. The most cisplatin-resistant subpopulation with the shortest doubling time was CD44-/CD90+, but this subpopulation had a low migratory capacity.


Prognostic values of tumor endothelial markers in patients with colorectal cancer.
Tumor endothelial markers (TEMs) are a newly discovered family of endothelial markers associated with tumor specific angiogenesis. This study sought to examine the levels of expression (qualitatively and quantitatively) for TEMs in human colon cancer. Human colorectal cancer tissues (n = 48) and normal background tissues (n = 31) were obtained after surgery. RNA was extracted from frozen sections for gene amplification. The expression of TEMs (TEM-1 to TEM-8) was assessed using RT-PCR and their transcript levels were determined using real-time-quantitative PCR (Q-RT-PCR). TEM-1 (P = 0.01), TEM-7 (P = 0.04), TEM-7R (P = 0.03), TEM-8 (P = 0.001) significantly raised in colon cancer tissues compared with the levels detected in normal background tissues. The expressions of TEM-2 and TEM-6 were found to be not significantly different between tumor tissues and normal tissues (P > 0.05). Patients who had cancer penetrating into and through the muscularis propria of the bowel wall and developed nodal involvement (Dukes C) exhibited significantly higher levels of TEM -8 compared to patients who were node negative (P < 0.05). TEM-7 and TEM-7R showed high level of transcripts in Dukes C, but they were not statistically significant. The level of the expression of TEM-1, TEM-7, TEM-7R and TEM-8 (but not TEM-2 and TEM-6) were associated with both nodal involvement and disease progression, and may therefore, have a prognostic value in colorectal cancer.


Paragliding injuries.
Regulations controlling the sport of paragliding were issued in April 1987 by the German Department of Transportation. The growing popularity of this sport has led to a steady increase in the number of associated injuries. This study presents the incidence, localization and degree of injuries associated with paragliding documented in Germany, Austria and Switzerland. The 283 injuries suffered by 218 paragliders were documented in the period 1987-1989: 181 occurred during landing, 28 during starting procedures and nine during flight. The mean patient age was 29.6 years. There were 34.9% spinal injuries, 13.4% upper extremity injuries and 41.3% lower limb injuries. Over half of these injuries were treated surgically and in 54 instances permanent disability remained. In paragliding the lower extremities are at greatest risk of injury during landing. Proper equipment, especially sturdy footwear, exact training in landing techniques as well as improved instruction in procedures during aborted or crash landings is required to reduce the frequency of these injuries.


Outpatient management of schizophrenia.
As effective antipsychotic pharmacotherapy has become available, patients with schizophrenia are increasingly managed in an outpatient setting by primary care physicians. Pharmacotherapy is generally effective in treating "positive," or psychotic, symptoms and lessening the risks of relapse, but ineffective in improving "negative," or deficit, symptoms. Aggressive attempts to totally control positive symptoms and to ameliorate negative symptoms tend to increase side effects and may be detrimental to the patient. Intensive psychotherapeutic and rehabilitative approaches are generally unproductive. Attempting to obtain a cure is unrealistic. A moderate approach is recommended, taking into consideration the limitations of existing treatments, achieving control of extreme symptoms and minimizing social and occupational limitations.


In vivo responses to inhaled proteins. III. Inhibition of experimental immune complex pneumonitis after suppression of peripheral blood lymphocytes.
We have previously shown that inhaled Con A has a powerful enhancing effect on the formation of immune complexes between an inhaled antigen and circulating antibody. Immunohistochemical staining has demonstrated such complexes, together with host complement, in close association with foci of necrotizing destruction of the pulmonary parenchyma. We have postulated that Con A promotes immune complex formation indirectly through polyclonal activation of lymphocytes in the lung. In this paper we test this hypothesis in animals rendered unresponsive to Con A stimulation in vivo by i.v. administration of cholera toxin (CT). Such treatment raised the levels of cAMP in peripheral blood lymphocytes and inhibited their proliferative response to Con A in vitro. CT administration further blocked the local inflammatory response to intradermal injections of Con A, as well as the cell-mediated immune response to intradermal injections of BSA. Although CT failed to block the immune complex-mediated Arthus vasculitis in the skin, it did block production of immune complex pulmonary injury by antigen/mitogen aerosols, as did decomplementation with purified cobra venom factor. These findings support the hypothesis that polyclonal activation of pulmonary lymphocytes promotes immune complex-type alveolitis, possibly by facilitating interactions between humoral antibody and intra-alveolar antigen.


BDNF and Krox20 as Indicators of Platelet-rich Plasma-Induced Nerve Regeneration in a Neuropathic Orofacial Pain Model.
Various growth factors contained in PRP can increase angiogenesis and cell proliferation, which plays an essential role in the process of neuroregeneration and peripheral nerve injury recovery. This study analyzed PRP effects in the neuro-regeneration of axonotmesis through brain-derived neurotrophic factor (BDNF) and Krox20 expressions. Freeze-dried allogeneic platelet-rich plasma (PRP) were prepared from allogeneic sources. Forty-two Rattus norvegicus were divided into three groups: negative control group, positive control group (crushing infraorbital nerve) and treatment group (crushing infraorbital nerve without PRP injection). Each group was observed for fourteen and twenty-one days after injury. Infraorbital nerve tissue is isolated for indirect immunohistochemistry examination with BDNF and Krox20 antibodies. Data analysis was performed using One-Way ANOVA and Mann-Whitney tests with significant value as p < 0.05. The PRP group showed BDNF expression significantly higher than control positive groups, both observation days (p = 0.00). A higher Korx20 expression showed by the PRP group after 21 days than in the control positive groups (p = 0.002). PRP can potentially improve neuroregeneration of axonotmesis through increased BDNF and Krox20 expression on the twenty-one days after injury.


Dose-dependent autophagic effect of titanium dioxide nanoparticles in human HaCaT cells at non-cytotoxic levels.
Interactions between nanoparticles and cells are now the focus of a fast-growing area of research. Though many nanoparticles interact with cells without any acute toxic responses, metal oxide nanoparticles including those composed of titanium dioxide (TiO2-NPs) may disrupt the intracellular process of macroautophagy. Autophagy plays a key role in human health and disease, particularly in cancer and neurodegenerative diseases. We herein investigated the in vitro biological effects of TiO2-NPs (18 nm) on autophagy in human keratinocytes (HaCaT) cells at non-cytotoxic levels. TiO2-NPs were characterized by transmission electron microscopy (TEM) and dynamic light scattering techniques. Cellular uptake, as evaluated by TEM and NanoSIMS revealed that NPs internalization led to the formation of autophagosomes. TiO2-NPs treatment did not reduce cell viability of HaCaT cells nor increased oxidative stress. Cellular autophagy was additionally evaluated by confocal microscopy using eGFP-LC3 keratinocytes, western blotting of autophagy marker LC3I/II, immunodetection of p62 and NBR1 proteins, and gene expression of LC3II, p62, NBR1, beclin1 and ATG5 by RT-qPCR. We also confirmed the formation and accumulation of autophagosomes in NPs treated cells with LC3-II upregulation. Based on the lack of degradation of p62 and NBR1 proteins, autophagosomes accumulation at a high dose (25.0 μg/ml) is due to blockage while a low dose (0.16 μg/ml) promoted autophagy. Cellular viability was not affected in either case. The uptake of TiO2-NPs led to a dose-dependent increase in autophagic effect under non-cytotoxic conditions. Our results suggest dose-dependent autophagic effect over time as a cellular response to TiO2-NPs. Most importantly, these findings suggest that simple toxicity data are not enough to understand the full impact of TiO2-NPs and their effects on cellular pathways or function.


Growing Role of Regorafenib in the Treatment of Patients with Sarcoma.
Sarcomas encompass a group of rare solid tumors responsible for approximately 1% of all cancer-related deaths in the United States each year. Subtypes include, but are not limited to, soft tissue sarcomas (STS) such as leiomyosarcoma, liposarcoma, pleomorphic sarcoma, and gastrointestinal stromal tumor (GIST). Treatment options for patients with STS vary depending on, among other factors, histological subtype. Data from a mix of phase 2 and phase 3 trials have suggested that the orally available multikinase inhibitor regorafenib may have efficacy in patients with STS who have progressed on previous lines of systemic therapy. Some clinical benefit of regorafenib has been shown in patients with leiomyosarcoma, synovial sarcoma, GIST, Ewing's sarcoma, and other sarcoma subtypes, suggesting a broad spectrum of potential activity in this population. Studies have also shown that the safety profile of regorafenib is acceptable in these patients, with adverse events that can be managed through dose reductions and/or interruptions as well as other supportive measures.


Is a share buyback right for your company?
Contrary to popular wisdom, buybacks don't create value by raising earnings per share. But they do indeed create value, and in two very different ways. First, a buyback sends signals about the company's prospects to the market--hopefully, that prospects are so good that the best investment managers can make right now is in their own company. But investors won't see it that way if other, negative, signals are coming from the company, and it's rarely a good idea for companies in high-growth industries, where investors expect that money to be spent pursuing new opportunities. Second, when financed as a debt issue, a buyback is essentially an exchange of equity for debt, conferring the traditional benefits of leverage--a tax shield and a discipline for managers. For such a buyback to make sense, a company would need to have taxable profits in need of shielding, of course, and be able to predict its future cash flows fairly accurately. Justin Pettit has found that managers routinely underestimate how many shares they need to buy to send a credible signal to the markets, and he offers a way to calculate that number. He also goes through the iterative steps involved in working out how many shares must be purchased to reach a target level of debt. Then he takes a look at the advantages and disadvantages of the three most common ways that companies make the actual purchases--open-market purchases, fixed-price tender offers, and auction-based tender offers. When a company's performance is lagging, a share buyback can look attractive. Unfortunately, a buyback can backfire--unless executives understand why, when, and how to use this powerful and risky tool.


Intrinsic Nanoscale Structure of Thin Film Composite Polyamide Membranes: Connectivity, Defects, and Structure-Property Correlation.
Transport of water, solutes, and contaminants through a thin film composite (TFC) membrane is governed by the intrinsic structure of its polyamide separation layer. In this work, we systematically characterized the nanoscale polyamide structure of four commercial TFC membranes to reveal the underlying structure-property relationship. For all the membranes, their polyamide layers have an intrinsic thickness in the range of 10-20 nm, which is an order of magnitude smaller than the more frequently reported apparent thickness of the roughness protuberances due to the ubiquitous presence of nanovoids within the rejection layers. Tracer filtration tests confirmed that these nanovoids are well connected to the pores in the substrates via the honeycomb-like opening of the backside of the polyamide layers such that the actual separation takes place at the frontside of the polyamide layer. Compared to SW30HR and BW30, loose membranes XLE and NF90 have thinner intrinsic thickness and greater effective filtration area (e.g., by the creation of secondary roughness features) for their polyamide layers, which correlates well to their significantly higher water permeability and lower salt rejection. With the aid of scanning electron microscopy, transmission electron microscopy, and tracer tests, the current study reveals the presence of nanosized defects in a polyamide film, which is possibly promoted by excessive interfacial degassing. The presence of such defects not only impairs the salt rejection but also has major implications for the removal of pathogens and micropollutants.


Topical microbicides--what's new?
Topical microbicides are an important, promising but complex HIV prevention technology under development. After 11 disappointing effectiveness trial outcomes of 6 candidate products (some tested as multiple doses and formulations) over the past 20 years, there is renewed optimism that a safe and effective microbicide will soon be available if the recent success of coitally linked use of the antiretroviral-based microbicide, 1% tenofovir gel, is confirmed. Studies of new antiviral agents, novel delivery mechanisms, and combination/multipurpose products that address challenges of adherence, enhance the effectiveness of tenofovir gel, and address sexual and reproductive health needs of men and women, including preventing HIV infection, are already underway.


The Use of Complementary and Alternative Medicine (CAM) in Psychiatric Units in Sweden.
Mental ill-health has been termed the pandemic of the 21st century, and a large share of those exposed do not receive treatment. Many people with depression, anxiety and other mental health problems consult complementary or alternative medicine (CAM), and CAM is used in conventional psychiatric care, in Sweden and in other countries. However, the extent to which CAM is used in psychiatric care, and for what purposes, are largely unknown. This study is based on a survey distributed to all heads of regional, municipal, private and governmental health care units treating persons with psychiatric symptoms across Sweden in 2019. CAM was reportedly used by 62% of the 489 responding health care units, for symptoms including anxiety, sleep disturbances and depression. Main motivations for CAM use were symptom relief, meeting patients' requests and reduced demand for pharmaceutical medication. Very few respondents reported side effects. The most common reason for interrupting CAM use at a unit was a lack of trained professionals. This study confirms the need for further research about CAM, and for CAM education and training among healthcare professionals.


A comparison of the changes in suprathreshold apparent contrast in acute and simulated optic neuritis.
A thin PVC film was used as a diffusion filter together with glass lenses in order to induce a unilateral artificial contrast sensitivity depression in controls. These artificially depressed contrast sensitivity functions were made comparable to those assessed in patients during the recovery phase of their acute unilateral optic neuritis. A technique of subjective suprathreshold contrast matching was used to determine the suprathreshold apparent contrast in patients and controls with acute and simulated optic neuritis respectively. The results showed that differences in apparent contrast between the eyes is proportional to the discrepancy in threshold contrast for high and intermediate spatial frequencies in patients with optic neuritis. For low frequencies the apparent contrast difference was independent of threshold differences. Contrast vision of controls with simulated optic neuritis did not show this frequency dependency. There was also a discrepancy in visual acuity between the two groups. Generally the control with artificially depressed contrast sensitivity functions has a letter acuity value lower than that of the optic neuritis patient with a corresponding CSF depression.


Polycyclic aromatic hydrocarbon and hypoxia exposures result in mitochondrial dysfunction in zebrafish.
Organisms are routinely subjected to a variety of environmental and chemical perturbations simultaneously. Often, multi-stressor exposures result in unpredictable toxicity that occurs through unidentified mechanisms. Here, we focus on polycyclic aromatic hydrocarbons (PAHs) and hypoxia, two environmental and physiological stressors that are known to co-occur in the environment. The aim of this study was to assess whether interactive mitochondrial dysfunction resulted from co-exposures of PAHs and hypoxia. Zebrafish embryos were co-exposed to non-teratogenic concentrations of an environmental PAH mixture and hypoxia beginning at 6 hpf for an acute period of 24 h and afterwards were given either no recovery period, 45 min, 5 -hs, or 18 -hs of recovery time in clean conditions. Mitochondrial function and integrity were assessed through the use of both in ovo and in vitro assays. Hypoxia exposures resulted in drastic reductions in parameters relating to mitochondrial respiration, ATP turnover, and mitochondrial DNA integrity. PAH exposures affected ATP production and content, as well as mitochondrial membrane dynamics and lactate content. While PAH and hypoxia exposures caused a broad range of effects, there appeared to be very little interaction between the two stressors in the co-exposure group. However, because hypoxia significantly altered mitochondrial function, the possibility remains that these effects may limit an individual's ability to respond to PAH toxicity and therefore could cause downstream interactive effects.


Drug-induced acute pancreatitis in a cohort of 328 patients. A single-centre experience from Australia.
Acute pancreatitis is associated with risk of morbidity and even mortality. Routine prescription drugs have been linked to the causation of acute pancreatitis. To determine the incidence, presentation, course and outcome of drug-induced acute pancreatitis amongst patients admitted to a public hospital. A retrospective analysis of patients presenting with acute pancreatitis to the Modbury Hospital, South Australia from January 2006 to April 2011. Each admission was reviewed within the electronic database for patient details as well as to determine the aetiological factor. In patients with drug-induced acute pancreatitis, the WHO Probability Scale was used to evaluate causality relationship. Three-hundreds and 28 patients were treated for acute pancreatitis during the study period. Biliary and alcohol-induced acute pancreatitis accounted for 80.8% of cases. Eleven patients (2 male and 9 female patients; median age: 59 years) were diagnosed with drug-induced acute pancreatitis. These included 5 cases of codeine-, 2 cases of azathioprine-, and 1 case each of chlorothiazide-, valproic acid-, oestradiol- and simvastatin-induced acute pancreatitis. Nine patients had a mild disease while 2 patients had severe acute pancreatitis with a median hospital stay of 4 days. Withdrawal of the drug resulted in cessation of the attacks in all patients over a median follow-up of 24 months. Routine prescription drugs, as an aetiological factor, accounted for 3.4% of cases of acute pancreatitis. The disease appeared to be more common in middle-aged women. It is likely that the overall incidence of this entity is under-reported owing to the stringent criteria needed to conclusively determine a causal relationship.


Looking through a different lens: Examining the inequality-mortality association in U.S. counties using spatial panel models.
Two areas still need further examination in the ecological study of inequality and mortality. First, the evidence for the relationship between income inequality and mortality remains inconclusive, particularly when the analytic unit is small (e.g., county in the U.S.). Second, most previous studies are cross-sectional and are unable to address the recent diverging patterns whereby mortality has decreased and income inequality increased. This study aims to contribute to both topic areas by studying the relationship between inequality and mortality via a spatiotemporal approach that simultaneously considers the spatial structure and the temporal trends of inequality and mortality using county panel data between 1990 and 2010 for the conterminous U.S. Using both spatial panel random effect model and spatial panel fixed effect models, we found that (a) income inequality was not a significant factor for mortality after taking into account the spatiotemporal structure and the most salient factors for mortality (e.g., socioeconomic status); (b) the spatial panel fixed effect model indicated that income inequality was negatively associated with mortality over the time, a relationship mirroring the diverging patterns; and (c) the significant spatial and temporal fixed effects suggested that both dimensions are critical factors in understanding the inequality-mortality relationship in the U.S. Our findings extend support to the argument that income inequality does not affect mortality and suggest that the cross-sectional findings may be a consequence of ignoring the temporal trends.


Acute and Subchronic Toxicity Study of Tamra Bhasma (Incinerated Copper) prepared from Ashodhita (Unpurified) and Shodhita (Purified) Tamra in Rats.
The use of metals in traditional medicines is very often seen as matter of concern these days, especially the Bhasma preparations which are always under stringent observations for containing highly reactive inorganic elements such as lead, mercury, arsenic and others. One of the Bhasma extensively used in routine Ayurvedic practice is Tamra (copper) bhasma. If it is not prepared properly or Shodhana procedure is not done properly, it acts as a poison. To indicate its toxic potential, Ashtamahadoshas (eight major ill effects) have been quoted in classics and due emphasis have been given to its Shodhana procedure. In the present study, Tamra bhasma prepared from Shodhita and Ashodhita Tamra was subjected to oral toxicity study to ascertain the role of Shodhana process on safety profile of Tamra bhasma on subchronic administration to albino rats. Both the samples were administered to rats for 45 consecutive days at the doses of 5.5, 27.5, and 55 mg/kg. Animals were sacrificed on 46(th) day and parameters like hematological, serum biochemical, and histopathology of various organs were studied. Results showed that Tamra bhasma prepared from Ashodhita Tamra has pathological implications on different hematological, serum biochemical and cytoarchitecture of different organs even at therapeutic dose level (5.5 mg/kg). Whereas, Tamra bhasma prepared from Shodhita Tamra is safe even at five-fold to therapeutic equivalent doses (27.5 mg/kg). These observations emphasize the role of Shodhana and importance of dose in expression of toxicity of the medicinal preparations.


Association of mutations in the Plasmodium falciparum Kelch13 gene (Pf3D7_1343700) with parasite clearance rates after artemisinin-based treatments-a WWARN individual patient data meta-analysis.
Plasmodium falciparum infections with slow parasite clearance following artemisinin-based therapies are widespread in the Greater Mekong Subregion. A molecular marker of the slow clearance phenotype has been identified: single genetic changes within the propeller region of the Kelch13 protein (pfk13; Pf3D7_1343700). Global searches have identified almost 200 different non-synonymous mutant pfk13 genotypes. Most mutations occur at low prevalence and have uncertain functional significance. To characterize the impact of different pfk13 mutations on parasite clearance, we conducted an individual patient data meta-analysis of the associations between parasite clearance half-life (PC1/2) and pfk13 genotype based on a large set of individual patient records from Asia and Africa. A systematic literature review following the PRISMA protocol was conducted to identify studies published between 2000 and 2017 which included frequent parasite counts and pfk13 genotyping. Four databases (Ovid Medline, PubMed, Ovid Embase, and Web of Science Core Collection) were searched. Eighteen studies (15 from Asia, 2 from Africa, and one multicenter study with sites on both continents) met inclusion criteria and were shared. Associations between the log transformed PC1/2 values and pfk13 genotype were assessed using multivariable regression models with random effects for study site. Both the pfk13 genotypes and the PC1/2 were available from 3250 (95%) patients (n = 3012 from Asia (93%), n = 238 from Africa (7%)). Among Asian isolates, all pfk13 propeller region mutant alleles observed in five or more specific isolates were associated with a 1.5- to 2.7-fold longer geometric mean PC1/2 compared to the PC1/2 of wild type isolates (all p ≤ 0.002). In addition, mutant allele E252Q located in the P. falciparum region of pfk13 was associated with 1.5-fold (95%CI 1.4-1.6) longer PC1/2. None of the isolates from four countries in Africa showed a significant difference between the PC1/2 of parasites with or without pfk13 propeller region mutations. Previously, the association of six pfk13 propeller mutant alleles with delayed parasite clearance had been confirmed. This analysis demonstrates that 15 additional pfk13 alleles are associated strongly with the slow-clearing phenotype in Southeast Asia. Pooled analysis associated 20 pfk13 propeller region mutant alleles with the slow clearance phenotype, including 15 mutations not confirmed previously.


Clinical effect of acupoint application with turmeric blistering moxibustion plaster on post-stroke hemiplegic shoulder pain.
To observe the effects of acupoint application with turmeric blistering moxibustion plaster on pain, shoulder range of motion (ROM) and upper limb motor function in the patients with post-stroke hemiplegic shoulder pain (PSHSP). Eighty-two patients with PSHSP were randomly divided into an observation group (41 cases, 1 case was eliminated, 4 cases dropped out) and a control group (41 cases, 2 cases were eliminated and 2 cases dropped out). The routine treatment, nursing care and rehabilitation training were performed in the control group. On the basis of the intervention as the control group, in the observation group, the turmeric blistering moxibustion plaster was applied to bilateral ashi points, Jianyu (LI 15), Jianliao (TE 14), Binao (LI 14), Shousanli (LI 10) and Hegu (LI 4), once a day, remained for 6 hours each time. This moxibustion therapy was operated 5 times weekly, one course of treatment consisted of 2 weeks and 2 courses were required. Separately, before treatment and after 2 and 4 weeks of treatment, the score of visual analogue scale (VAS), shoulder ROM and the score of upper limbs in Fugl-Meyer assessment (U-FMA) were observed in the two groups. VAS scores were lower (P<0.05), ROM in shoulder flexion, abduction, internal rotation and external rotation was larger (P<0.05), and U-FMA scores were higher (P<0.05) after 2 and 4 weeks of treatment when compared with those before treatment in the two groups. After 4 weeks of treatment, VAS score decreased (P<0.05), and ROM in shoulder flexion, abduction, internal rotation, external rotation and U-FMA score increased (P<0.05) in comparison with those after 2 weeks of treatment in either group. In the observation group, VAS scores were dropped (P<0.05) after 2 and 4 weeks of treatment respectively, and ROM of shoulder flexion and abduction enlarged after 2 weeks of treatment (P<0.05) when compared with those in the control group. After 4 weeks of treatment, ROM in shoulder flexion, abduction, internal rotation and external rotation in the observation group was larger (P<0.05) and U-FMA score was higher (P<0.05) than those in the control group. Acupoint application with turmeric blistering moxibustion plaster may effectively reduce the degree of shoulder pain and improve the shoulder range of motion and the upper limb motor function in the patients with post-stroke hemiplegic shoulder pain.


[Analysis of parameters of the immunoreactivity curve].
The analysis of the general principles of interaction between the immune systems of macroorganisms and the causative agents of infectious diseases helped to describe the mathematical model of the vaccinal process. This model can be used for standardization of criteria for the evaluation of vaccinal preparations in accordance with their immunogenicity, reactogenicity and and safety.


Lysyl-tRNA synthetase produces diadenosine tetraphosphate to curb STING-dependent inflammation.
Inflammation is an essential part of immunity against pathogens and tumors but can promote disease if not tightly regulated. Self and non-self-nucleic acids can trigger inflammation, through recognition by the cyclic GMP-AMP (cGAMP) synthetase (cGAS) and subsequent activation of the stimulator of interferon genes (STING) protein. Here, we show that RNA:DNA hybrids can be detected by cGAS and that the Lysyl-tRNA synthetase (LysRS) inhibits STING activation through two complementary mechanisms. First, LysRS interacts with RNA:DNA hybrids, delaying recognition by cGAS and impeding cGAMP production. Second, RNA:DNA hybrids stimulate LysRS-dependent production of diadenosine tetraphosphate (Ap4A) that in turn attenuates STING-dependent signaling. We propose a model whereby these mechanisms cooperate to buffer STING activation. Consequently, modulation of the LysRS-Ap4A axis in vitro or in vivo interferes with inflammatory responses. Thus, altogether, we establish LysRS and Ap4A as pharmacological targets to control STING signaling and treat inflammatory diseases.


Herpes genitalis and circumcision.
214 patients with genital herpes infection proven by culture and a control group of 410 other patients were included in a retrospective study devised to investigate the relationship of circumcision to genital HSV infection in the male. The percentage of patients circumicised in the control group was significantly larger than in the herpes group (P less than 0-01). The was not found to be the case for other sexually-transmitted diseases apart from monilial balanitis. There was also a significant difference in contraceptive methods in the two groups, barrier methods being used less ofter than other methods in the herpes group and the reverse in the control group. It is concluded that there is a positive relationship between absence of circumicision and genital HSV infection, but that a prospective study should be undertaken to confirm these results.


Neutralizing TIMP1 restores fecundity in a rat model of endometriosis and treating control rats with TIMP1 causes anomalies in ovarian function and embryo development.
Human and rat endometriotic lesions synthesize and secrete tissue inhibitor of metalloproteinase 1 (TIMP1). More TIMP1 localizes in the ovarian theca in an established rat model for endometriosis (Endo) when compared to surgical controls (Sham). We hypothesized that endometriotic TIMP1 secreted into peritoneal fluid (PF) negatively affects ovarian function and embryogenesis by altering the balance of matrix metalloproteinases (MMPs) and TIMPs. Three experiments were performed modulating TIMP1 in vitro and in vivo to investigate ovarian and embryonic anomalies. The first experiment demonstrated control embryos treated in vitro with endometriotic PF concentrations of TIMP1 developed abnormally. In the second experiment where TIMP1 was modulated in vivo, TIMP1-treated Sham rats had fewer zygotes, ovarian follicles, and corpora lutea (CLs) and poorer embryo quality and development, which is analogous to the findings in Endo rats. Importantly, Endo rats treated with a TIMP1 function-blocking antibody had zygote, follicle, and CL numbers and embryo quality similar to Sham rats. In addition, more TIMP1 inhibitory activity was found in ovaries from Endo and TIMP1-treated Sham rats than in ovaries from Sham or TIMP1 antibody-treated Endo rats. In experiment three, control rats (no surgery) treated with Endo PF had fewer follicles and CLs and increased TIMP1 localization in the ovarian theca whereas treatment with Endo PF stripped of TIMP1 or with Sham PF had no effect, providing further evidence that endometriotic TIMP1 sequesters in the ovary and inhibits MMPs necessary for ovulation. Collectively, these results showed that excessive TIMP1 was deleterious to ovulation and embryo development. Thus, novel TIMP1-modulating therapies may be developed to alleviate infertility in women with endometriosis.


Utility intervention programs in the evolutional control of patients with heart failure
Acute heart failure (HF) is a prevalent disease with important socio-economic repercussions. Due to the aging of population, these values will increase in the coming years, so it may be useful to the implementation of intervention programs in these patients to decrease morbidity and mortality. A quasi-experimental prospective study (n = 262) of patients admitted at the Internal Medicine Department of the Hospital Clínico Universitario Lozano Blesa, in Zaragoza, Spain, diagnosed of HF between November 2013 and October 2014 (both dates inclusive) (n = 108) followed up for 1 year was performed. Within this group, a subgroup with an intensive intervention (n = 30) was performed. The data were compared with a historical cohort of patients admitted to the same department during the same time in the previous year (from November 2012 to October 2013) (n = 154). Statistically significant differences between groups attending to the therapeutical adherence to clinical guidelines (p < 0.011) were observed. Considering the intensive intervention subgroup, statistically significant differences were observed in the rate of exitus (p < 0.032) and survival (log rank <0.030) compared to the control group. The close monitoring of patients with HF improves adherence, reduces mortality and improves survival. This May result in a decline in the use of health resources, which entails significant socio-economic benefits.


Dependence of both spontaneous and antibody-dependent, granule exocytosis-mediated NK cell cytotoxicity on extracellular signal-regulated kinases.
Extracellular signal-regulated kinases (ERK, also known as mitogen-activated protein kinases) are serine-threonine kinases transducing signals elicited upon ligand binding to several tyrosine kinase-associated receptors. We have reported that ERK2 phosphorylation and activation follows engagement of the low affinity receptor for the Fc portion of IgG (CD16) on NK cells, and is necessary for CD16-induced TNF-alpha mRNA expression. Here, we analyzed the involvement of ERK in NK cell-mediated cytotoxicity and IFN-gamma expression induced upon stimulation with targets cells, coated or not with Abs. Our data indicate that, as with immune complexes, ERK2 phosphorylation occurs in human primary NK cells upon interaction with target cells sensitive to granule exocytosis-mediated spontaneous cytotoxicity, and that this regulates both target cell- and immune complex-induced cytotoxicity and IFN-gamma mRNA expression. A specific inhibitor of mitogen-activated protein kinase kinase reduced both spontaneous and Ab-dependent cytotoxicity in a dose-dependent manner involving, at least in part, inhibition of granule exocytosis without affecting effector/target cell interaction and rearrangement of the cytoskeleton proteins actin and tubulin. Involvement of ERK in the regulation of Ca2+-dependent cell-mediated cytotoxicity was confirmed, using a genetic approach, in primary NK cells infected with a recombinant vaccinia virus encoding an ERK inactive mutant. These data indicate that the biochemical pathways elicited in NK cells upon engagement of receptors responsible for either spontaneous or Ab-dependent recognition of target cells, although distinct, utilize ERK as one of their downstream molecules to regulate effector functions.


Three decades of reimplantation of the aortic valve-the Brussels experience.
Over the last three decades, the importance of native valve preservation has increasingly become evident. Valve-sparing root replacement procedures, such as the reimplantation or remodeling technique, are therefore being progressively used for aortic root replacement and/or aortic valve repair. Herein, we are summarizing our single-center experience with the reimplantation technique. We queried our prospective database for aortic valve repair and recruited all adult (≥18 years) patients who have undergone valve-sparing root replacement with the reimplantation technique between March 1998 and January 2022. We subcategorized the patients into three distinct groups: root aneurysm without aortic regurgitation (AR) (grade ≤1+), root aneurysm with AR (grade >1+) and isolated chronic AR (root <45 mm). Univariable logistic regression analysis was performed to identify variables of interest, which were further analyzed by multivariable Cox-regression analysis. Survival, freedom from valve reintervention, and freedom from recurrent regurgitation, were analyzed with the Kaplan-Meier method. A total of 652 patients were recruited for this study; 213 patients underwent reimplantation for aortic aneurysm without AR, 289 patients for aortic aneurysm with AR, and 150 patients with isolated AR. Cumulative survival was 95.4% (95% CI: 92.9-97.0%) after 5 years, 84.8% (80.0-88.5%) after 10 years, and 79.5% (73.3-84.5%) after 12 years, which was comparable to the age-matched Belgian population. Older age (HR 1.06, P≤0.001) and male gender (HR 2.1, P=0.02) were associated with late mortality. Freedom from reoperation on the aortic valve at 5 years was 96.2% (95% CI: 93.8-97.7%), and 90.4% (95% CI: 87.4-94.2%) at 12 years. Age (P=0.001) and preoperative left ventricular end-diastolic dimension (LVEDD) (P=0.03) were associated with late reoperation. Our long-term data supports our reimplantation approach as a viable option for aortic root aneurysms and/or aortic regurgitation, with long-term survival that mirrors that of the general population.


Glycemic index and heart disease.
A diet high in carbohydrates with high glycemic indexes (GI) and glycemic load were linked to risk of coronary heart disease development in women in a large prospective study. Two cross-sectional studies showed that low-GI diets are associated with high HDL-cholesterol concentrations, especially in women. In a tightly controlled study of patients with type 2 diabetes, serum total cholesterol, LDL cholesterol, and apolipoprotein B concentrations fell more significantly after a low-GI diet than after a high-GI diet. In the same study, plasminogen activator inhibitor-1 concentrations were reduced by 58% after the low-GI diet. Insulin-stimulated glucose uptake by adipocytes was significantly higher in patients undergoing coronary artery bypass graft surgery after 4 wk of consuming a low-GI diet than after consuming a high-GI diet. The effects of low-GI diets may be mediated by changes in postprandial fatty acid concentrations or by hormonal signals from adipocytes, but a possible association of low-GI diets with some other dietary factor such as chromium must not be excluded. Proof of the clinical value of low-GI diets awaits prospective trials, which should include short-term observations covering periods of metabolic stress induced by surgery as well as long-term trials with clinical endpoints.


Differential regulation of formyl peptide receptor-like 1 expression during the differentiation of monocytes to dendritic cells and macrophages.
Monocytes are the common precursors for myeloid dendritic cells (DC) and macrophages. Identification of chemotactic receptors expressed by myeloid DC, macrophages, and their precursors in the course of differentiation and maturation is important not only for elucidation of their in vivo trafficking, but also for understanding of the functional distinction between DC and macrophages. We chose to study formyl peptide receptor like-1 (FPRL1), a chemotactic receptor known to interact with several endogenous agonists that are involved in inflammatory and host defense responses. Here we show that FPRL1 is down-regulated as monocytes differentiate into DC. This down-regulation occurs at both mRNA and functional levels. Therefore, the interaction of FPRL1 with its agonists is more likely to regulate the in vivo trafficking of DC precursors than DC. In contrast, FPRL1 expression is maintained at both mRNA and functional levels as monocytes differentiate into macrophages. Thus, our results demonstrate further distinctions between myeloid DC and macrophages, albeit they share a common precursor. The fact that macrophages rather than myeloid DC express functional FPRL1 suggests that this chemotactic receptor may be more involved in inflammatory reactions and innate host defense than in adaptive immune responses.


Host immune response to cytomegalovirus: products of transfected viral immediate-early genes are recognized by cloned cytolytic T lymphocytes.
To confirm that immediate-early (IE) genes of murine cytomegalovirus (MCMV) give rise to antigens recognized by specific cytolytic T lymphocytes (CTL), a 10.8-kilobase fragment of MCMV DNA which is abundantly transcribed at IE times was transfected into L cells expressing the Ld class I major histocompatibility glycoprotein. The viral genome fragment contains sequences of the three IE transcription units of MCMV: ie1, ie2, and ie3. In the transfected cell lines, only the predominant 2.75-kilobase transcript of ie1 and its translation product pp89 could be detected. The transfectants were analyzed for membrane expression of an IE antigen by employing clone IE1, an IE-specific CTL clone, as the probe. Only cells that expressed both the MCMV IE gene(s) and the Ld gene were recognized by the CTL clone.


MRI tools for assessment of microstructure and nephron function of the kidney.
MRI can provide excellent detail of renal structure and function. Recently, novel MR contrast mechanisms and imaging tools have been developed to evaluate microscopic kidney structures including the tubules and glomeruli. Quantitative MRI can assess local tubular function and is able to determine the concentrating mechanism of the kidney noninvasively in real time. Measuring single nephron function is now a near possibility. In parallel to advancing imaging techniques for kidney microstructure is a need to carefully understand the relationship between the local source of MRI contrast and the underlying physiological change. The development of these imaging markers can impact the accurate diagnosis and treatment of kidney disease. This study reviews the novel tools to examine kidney microstructure and local function and demonstrates the application of these methods in renal pathophysiology.


Pharmacodynamic Studies of Fluorescent Diamond Carriers of Doxorubicin in Liver Cancer Cells and Colorectal Cancer Organoids.
We recently reported on preferential deposition of bare fluorescent diamond particles FDP-NV-700/800nm (FDP-NV) in the liver following intravenous administration to rats. The pharmacokinetics of FDP-NV in that species indicated short residency in the circulation by rapid clearance by the liver. Retention of FDP-NV in the liver was not associated with any pathology. These observations suggested that cancer therapeutics, such as doxorubicin, linked to FDP-NV, could potentially serve for anti-cancer treatment while sparing toxicities of peripheral organs. To generate proof-of-concept (POC) and detail mechanisms of action of doxorubicin-coated FDP-NV-700/800nm (FDP-DOX) as a prospective chemotherapeutic for metastatic liver cancer. FDP-DOX was generated by adsorption chemistry. Experimental design included concentration and time-dependent efficacy studies as compared with naïve (baren) FDP-NV in in vitro liver cancer cells models. Uptake of FDP-NV and FDP-DOX by HepG-2, Hep-3B and hCRC organoids were demonstrated by flow-cytometry and fluorescent microscopy. FDP-DOX pharmacodynamic effects included metabolic as well as cell death biomarkers Annexin V, TUNEL and LDH leakage. DOX desorpted from FDP-DOX was assessed by confocal microscopy and chemical assay of cells fractions. FDP-DOX efficacy was dose- and time-dependent and manifested in both liver cancer cell lines and human CRC organoids. FDP-DOX was rapidly internalized into cancer cells/organoids leading to cancer growth inhibition and apoptosis. FDP-DOX disrupted cell membrane integrity as evident by LDH release and suppressing mitochondrial metabolic pathways (AlamarBlue assay). Access of free DOX to the nuclei was confirmed by direct UV-Visible fluorescent assay and confocal microscopy of DOX fluorescence. The rapid uptake and profound cancer inhibition observed using FDP-DOX in clinically relevant cancer models, highlight FDP-DOX promise for cancer chemotherapeutics. We also conclude that the in vitro data justify further investment in in vivo POC studies.


Accelerated Corneal Cross-linking as an Adjunct Therapy in the Management of Presumed Bacterial Keratitis: A Cohort Study.
To compare the outcomes of accelerated photoactivated chromophore for keratitis corneal cross-linking (PACK-CXL) as an adjunct treatment for bacterial keratitis (PACK-CXL plus standard antibiotic therapy) for patients receiving only standard antibiotic therapy. Retrospective cohort study of outcomes of patients with moderate infectious presumed bacterial keratitis (ulcer diameter 2 to 7 mm and stromal depth < 300 µm) were compared before and after initiation of a new treatment protocol of PACK-CXL in addition to standard antibiotic treatment. A total of 70 eyes of 70 patients were included: 39 eyes in the PACK-CXL plus antibiotic (PACK-ABX) group and 31 eyes in the antibiotic only (ABX) control group. The PACK-ABX group showed shorter times to complete reepithelialization (9.3 ± 6.0 vs 16.0 ± 12.7 days, P = .01) and did not require tectonic emergency keratoplasty (0% versus 19.4%, P = .006). The PACK-ABX group also showed a higher percentage of eyes with complete reepithelialization in 6 days or less (46.2% vs 6.5%, P < .001) and a trend for shorter hospitalizations (6.3 ± 5.0 vs 8.5 ± 4.5 days, P = .06). A multivariate analysis controlling for age showed that PACK-ABX treatment remained significantly associated with early ulcer reepithelialization (odds ratio = 0.09, 95% confidence interval = 0.02 to 0.48, P = .005). This study validates previous findings regarding the use of accelerated PACK-CXL in the treatment of bacterial keratitis. Adding PACK-CXL improved clinical outcomes (reducing healing time) when compared to antibiotics alone. [J Refract Surg. 2020;36(4):258-264.].


Performance of mixed effects models in the analysis of mediated longitudinal data.
Linear mixed effects models (LMMs) are a common approach for analyzing longitudinal data in a variety of settings. Although LMMs may be applied to complex data structures, such as settings where mediators are present, it is unclear whether they perform well relative to methods for mediational analyses such as structural equation models (SEMs), which have obvious appeal in such settings. For some researchers, SEMs may be more difficult than LMMs to implement, e.g. due to lack of training in the methodology or the need for specialized SEM software. It therefore is of interest to evaluate whether the LMM performs sufficiently in a scenario particularly suitable for SEMs. We focus on evaluation of the total effect (i.e. direct and indirect) of an exposure on an outcome of interest when a mediating factor is present. Our aim is to explore whether the LMM performs as well as the SEM in a setting that is conducive to using the SEM. We simulated mediated longitudinal data from an SEM where a binary, main independent variable has both direct and indirect effects on a continuous outcome. We conducted analyses with both the LMM and SEM to evaluate the performance of the LMM in a setting where the SEM is expected to be preferable. Models were evaluated with respect to bias, coverage probability and power. Sample size, effect size and error distribution of the simulated data were varied. Both models performed well in a range of settings. Marginal increases in power estimates were observed for the SEM, although generally there were no major differences in performance. Power for both models was good with a sample of size of 250 and a small to medium effect size. Bias did not substantially increase for either model when data were generated from distributions that were both skewed and kurtotic. In settings where the goal is to evaluate the overall effects, the LMM excluding mediating variables appears to have good performance with respect to power, bias and coverage probability relative to the SEM. The major benefit of SEMs is that it simultaneously and efficiently models both the direct and indirect effects of the mediation process.


Medicare skilled nursing facility reimbursement and upcoding.
Post-acute care provided by skilled nursing facilities (SNFs) is reimbursed by Medicare under a prospective payment system using resource utilization groups (RUGs) that adjust payment intensity on the basis of predefined ranges of weekly therapy minutes provided and the functionality of the patient. Individual RUGs account for differences in the intensity of care provided, but there exists significant regional variation in the payments SNFs receive from Medicare due to the use of geographic adjustment factors. This paper is the first to use this geographic variation in the generosity of Medicare reimbursement to empirically test if SNFs respond to payment differences between RUG categories. The results are highly suggestive that SNFs upcode patients by providing additional therapy minutes to increase revenue, whereas we find no evidence of upcoding related to patient functionality scores. Simulating how different payment differentials affect RUG selection, we predict that reducing the financial incentive to upcode could result in significant savings to Medicare.


Tattooing or not? A review of current practice and outcomes for laparoscopic colonic resection following endoscopy at a tertiary care centre.
Because small colonic tumours may not be visualized or palpated during laparoscopy, location of the lesion must be identified before surgery. The aim of this study was to evaluate the effectiveness of the current recommendation of endoscopic tattooing of lesions prior to laparoscopic colonic resections. All consecutive patients who underwent elective laparoscopic resection for a colonic lesion at a single tertiary institution between 2013 and 2015 were identified for chart review. In total, 224 patients underwent laparoscopic resection for a benign or malignant colonic lesion during the study period. All patients had a complete colonoscopy preoperatively. In all, 148 patients (66%) had their lesion tattooed at endoscopy. Most lesions were tattooed distally, but 15% were tattooed either proximally, both proximally and distally, or tattooed without specifying location as proximal or distal. Tattoo localization was accurate in 69% of cases. Tattooed lesions were not visible during surgery 21.5% of time; 2 cases were converted to open surgery to identify the lesion. Inaccuracy in endoscopic localization led to change in surgical plan in 16% of surgeries. In the nontattooed group, 1 case was converted to open surgery to localize the lesion, 3 required intraoperative colonoscopy and 1 had positive margins on final pathology. To improve surgical planning, we recommend the practice of endoscopic tattooing of all colon lesions at a location just distal to the lesion using multiple injections to cover the circumference of the bowel wall.


Rubus fruticosus leaf extract inhibits vascular dementia-induced memory impairment and neuronal loss by attenuating neuroinflammation.
Vascular dementia (VaD) is characterized by progressive memory impairment, which is associated with microglia-mediated neuroinflammation. Polyphenol-rich natural plants, which possess anti-inflammatory activities, have attracted scientific interest worldwide. This study investigated whether Rubus fruticosus leaf extract (RFLE) can attenuate VaD. Sprague-Dawley rats were separated into five groups: SO, sham-operated and treated with vehicle; OP, operated and treated with vehicle; RFLE-L, operated and treated with low dose (30 mg/kg) of RFLE; RFLE-M, operated and treated with medium dose (60 mg/kg) of RFLE; and RFLE-H, operated and treated with high dose (90 mg/kg) of RFLE. Bilateral common carotid artery and hypotension were used as a modeling procedure, and the RFLE were intraorally administered for 5 days (preoperative 2 and postoperative 3 days). The rats then underwent memory tests including the novel object recognition, Y-maze, Barnes maze, and passive avoidance tests, and neuronal viability and neuroinflammation were quantified in their hippocampi. The results showed that the OP group exhibited VaD-associated memory deficits, neuronal death, and microglial activation in hippocampi, while the RFLE-treated groups showed significant attenuation in all above parameters. Next, using BV-2 microglial cells challenged with lipopolysaccharide (LPS), we evaluated the effects of RFLE in dynamics of proinflammatory mediators and the upstream signaling pathway. RFLE pretreatment significantly inhibited the LPS-induced release of nitric oxide, TNF-α, and IL-6 and upregulation of the MAPKs/NF-κB/iNOS pathway. Collectively, we suggest that RFLE can attenuate the histologic alterations and memory deficits accompanied by VaD, and these roles are, partly due to the attenuation of microglial activation.


Automated calculation of symmetry measure on clinical photographs.
The quality of life of breast cancer survivors is ameliorated by minimizing adverse effects on their physical appearance. Breast reconstruction is important for restoring the survivor's appearance. In breast reconstructive surgery, there is a need to develop technologies for quantifying surgical outcomes and understanding women's perceptions of changes in their appearance. Methods for objectively measuring breast anatomy are needed in order to help breast cancer survivors, radiation oncologists and surgeons quantify changes in appearance that occur with different breast reconstructive surgical options. In this study, we present an automated method for computing a variant of the normalized Breast Retraction Assessment, a common measure of symmetry, from routine clinical photographs taken to document breast cancer treatment procedures. The algorithms were designed using a development set of retrospectively collected clinical photographs and evaluated using a separate test set of prospectively collected clinical photographs. We have developed new algorithms for automatically localizing the umbilicus and nipples on standard anterior-posterior clinical photographs, enabling the automatic calculation of measure of breast symmetry. This study demonstrates that under standardized photographic conditions, automatic localization of fiducial points and subsequent computation of measures, such as symmetry, is feasible. The algorithms presented here for quantifying surgical outcomes in breast reconstructive surgery will provide a foundation for future work on assisting a cancer patient and her surgeons in selecting and planning reconstruction procedures that will maximize the woman's psychosocial adjustment to life as a breast cancer survivor.


Factors released by rat type 1 astrocytes exert different effects on the proliferation of human neuroblastoma cells (SH-SY5Y) in vitro.
Brain metastases derived from abdominal neuroblastoma are an uncommon complication of this tumour; however, an increase in their occurrence has recently been reported. In the present study, we have investigated the influence of factors derived from central nervous system glial cells on the proliferation of human neuroblastoma cells (SH-SY5Y) in vitro. Co-culture experiments show that a 24-h exposure to factors released by type 1 astrocytes (A1) may induce a significant decrease in [(3)H]thymidine ([(3)H]TdR) incorporation by SH-SY5Y cells. This effect was not duplicated by fresh A1-conditioned medium (A1-CM); A1-CM became active only when it was heated or frozen. In contrast to this short-lived inhibitory effect, long-term treatment (3, 6 and 9 days) with A1-CM produced a significant and dose-dependent increase in SH-SY5Y cell number. Immunoneutralisation of A1-CM with an anti-transforming growth factor-beta antibody eliminated the inhibitory effect on [(3)H]TdR uptake in SH-SY5Y cells, but did not affect the increased number of viable cells observed after long-term treatments. In conclusion, these results showed that factor(s) released by A1 may affect the proliferation/survival of a human neuroblastoma cell line in vitro inducing: (a) a short transient negative effect on DNA synthesis and (b) an overall sustained trophic action. These results are suggestive of a possible role of glial cells in the establishment of brain metastases of neuroblastomas.


Distinct domains of mouse dishevelled are responsible for the c-Jun N-terminal kinase/stress-activated protein kinase activation and the axis formation in vertebrates.
Recent studies have shown that Drosophila Dishevelled (Dsh), an essential component of the wingless signal transduction, is also involved in planar polarity signaling through the c-Jun N-terminal kinase (JNK)/stress-activated protein kinase (SAPK) pathway in Drosophila. Here, we show that expression of a mouse homolog of Dsh (mDvl-1) in NIH3T3 cells activates JNK/SAPK, and its activator MKK7. A C-terminal half of mDvl-1 which contains the DEP domain was sufficient for the activation of JNK/SAPK, whereas an N-terminal half of mDvl-1 as well as the DEP domain is required for stimulation of the TCF/LEF-1-dependent transcriptional activation, a beta-catenin-dependent process. A single amino acid substitution (Met for Lys) within the DEP domain (mDvl-1 (KM)) abolished the JNK/SAPK-activating activity of mDvl-1, but did not affect the activity to activate the LEF-1-dependent transcription. Ectopic expression of mDvl-1 (KM) or an N-terminal half of mDvl-1, but not the C-terminal, was able to induce secondary axis in Xenopus embryos. Because the secondary axis formation is dependent on the Wnt/beta-catenin signaling pathway, these results suggest that distinct domains of mDvl-1 are responsible for the two downstream signaling pathways, the beta-catenin pathway and the JNK/SAPK pathway in vertebrates.


Sporotrichoid metastases to the skin from cutaneous squamous cell carcinoma in an immunocompetent patient.
Squamous cell carcinoma (SCC) of the skin may metastasize in more than 5% of cases. Metastases usually involve the primary regional lymph nodes in 85% of cases. The skin is rarely involved by metastases of SCC of the skin. The presence of multiple cutaneous metastases in a figurate array is a very rare occurrence, mostly reported in SCC occurring in immunocompromized patients. The case of a 70-year-old immunocompetent patient, with an ulcerated primary SCC on his right palm and 3 nodules in a linear array on his right arm is described. He also presented with homolateral axillary lymphadenopathy. Histopathologic examination from the 3 nodules revealed SCC of variable grading, with focal fusion of the metastases with the overlying epidermis. The patient had been previously treated with chemotherapy and radiotherapy in another Department, with no benefit. Despite right arm amputation and homolateral axillary lymphadenectomy, the patient died 4 months after a procedure, for pulmonary embolism.


Day-case versus overnight stay in laparoscopic cholecystectomy.
Although day-case elective laparoscopic cholecystectomy can save bed costs, its safety remains to be established. To assess the safety and benefits of day-case surgery compared to overnight stay in patients undergoing elective laparoscopic cholecystectomy. We searched The Cochrane Hepato-Biliary Group Controlled Trials Register, the Cochrane Central Register of Controlled Trials (CENTRAL) in The Cochrane Library, MEDLINE, EMBASE, and Science Citation Index Expanded until February 2007 for identifying randomised trials using search strategies. Only randomised clinical trials, irrespective of language, blinding, or publication status, comparing day-case and overnight stay in elective laparoscopic cholecystectomy were considered for the review. We collected the data on the characteristics of the trial, methodological quality of the trials, morbidity, prolonged hospitalisation, re-admissions, pain and quality of life from each trial. We analysed the data with both the fixed-effect and the random-effects models using RevMan Analysis. For each outcome we calculated the relative risk, weighted mean difference, or standardised mean difference with 95% confidence intervals (CI) based on available case-analysis. Five trials with 429 patients randomised to the day-case group (215) and overnight stay group (214) were included in the review. Four of the five trials were of low risk of bias regarding randomisation and follow up, but all lacked blinding. The trials recruited 49% of patients undergoing cholecystectomy. The selection criteria varied, but most included only patients without other diseases. The patients were living in easy reach of the hospital and with a responsible adult to take care of them. On the day of surgery, 81% of day-case patients were discharged. The drop-out rate after randomisation varied from 6.5% to 12.7%. There was no significant difference between day-case and overnight stay group as regards to morbidity, prolongation of hospital stay, re-admission rates, pain, quality of life, patient satisfaction and return to normal activity and work. Day-case elective laparoscopic cholecystectomy seems to be a safe and effective intervention in selected patients (with no or minimal systemic disease and within easy reach of the hospital) with symptomatic gallstones. Because of the decreased hospital stay, it is likely to save costs.


Intra-operative administration of low-dose IV glucose attenuates post-operative insulin resistance.
Insulin sensitivity often decreases after surgery in spite of normal insulin secretion, and may worsen the outcome. This post-operative insulin resistance increases according to the magnitude of surgical invasion. However, supplementation of carbohydrates before surgery attenuates the post-operative insulin resistance. This study aimed to investigate the effect of intra-operative administration of low-dose glucose on the post-operative insulin resistance. Patients undergoing maxillofacial surgery were randomly assigned to two groups throughout the surgical procedure: The glucose group receiving acetated Ringer solution with 1.5% glucose and the control group receiving acetated Ringer solution without glucose. Insulin resistance quantified by the mean glucose infusion rate (the glucose infusion rate) was evaluated by glucose clamp using the STG-22TM instrument on the previous day and on the next day of surgery. Blood glucose level was monitored continuously during surgery. In addition, serum insulin, ketone bodies and 3-methylhistidine were measured during perioperative period. Patients in the glucose group (n=11) received 0.15 ± 0.06 g/kg/h of glucose during surgery, while patients in the control group (n=11) received no glucose. In both groups, however, the mean blood glucose levels were maintained stable at less than 150 mg/dL during and after surgery. The serum ketone bodies significantly increased after surgery in the control group (p=0.0035), while it decreased significantly in the glucose group (p=0.043). The reduction rate in the glucose infusion rate was significantly lower in the glucose group, 43.3 ± 20.7%, than that in the control group, 57.7 ± 9.3% (p=0.041). Intra-operative small-dose of glucose administration may suppress ketogenesis and attenuate the post-operative insulin resistance without causing hyperglycemia.


Immune-mediated hookworm clearance and survival of a marine mammal decrease with warmer ocean temperatures.
Increases in ocean temperature are associated with changes in the distribution of fish stocks, and the foraging regimes and maternal attendance patterns of marine mammals. However, it is not well understood how these changes affect offspring health and survival. The maternal attendance patterns and immunity of South American fur seals were assessed in a rookery where hookworm disease is the main cause of pup mortality. Pups receiving higher levels of maternal attendance had a positive energy balance and a more reactive immune system. These pups were able to expel hookworms through a specific immune mediated mechanism and survived the infection. Maternal attendance was higher in years with low sea surface temperature, therefore, the mean hookworm burden and mortality increased with sea surface temperature over a 10-year period. We provide a mechanistic explanation regarding how changes in ocean temperature and maternal care affect infectious diseases dynamics in a marine mammal.


Crossover from 2D to 3D in a weakly interacting Fermi gas.
We have studied the transition from two to three dimensions in a low temperature weakly interacting 6Li Fermi gas. Below a critical atom number N(2D) only the lowest transverse vibrational state of a highly anisotropic oblate trapping potential is occupied and the gas is two dimensional. Above N(2D) the Fermi gas enters the quasi-2D regime where shell structure associated with the filling of individual transverse oscillator states is apparent. This dimensional crossover is demonstrated through measurements of the cloud size and aspect ratio versus atom number.


Identification of ion-channel modulators that protect against aminoglycoside-induced hair cell death.
Aminoglycoside antibiotics are used to treat life-threatening bacterial infections but can cause deafness due to hair cell death in the inner ear. Compounds have been described that protect zebrafish lateral line hair cells from aminoglycosides, but few are effective in the cochlea. As the aminoglycosides interact with several ion channels, including the mechanoelectrical transducer (MET) channels by which they can enter hair cells, we screened 160 ion-channel modulators, seeking compounds that protect cochlear outer hair cells (OHCs) from aminoglycoside-induced death in vitro. Using zebrafish, 72 compounds were identified that either reduced loading of the MET-channel blocker FM 1-43FX, decreased Texas red-conjugated neomycin labeling, or reduced neomycin-induced hair cell death. After testing these 72 compounds, and 6 structurally similar compounds that failed in zebrafish, 13 were found that protected against gentamicin-induced death of OHCs in mouse cochlear cultures, 6 of which are permeant blockers of the hair cell MET channel. None of these compounds abrogated aminoglycoside antibacterial efficacy. By selecting those without adverse effects at high concentrations, 5 emerged as leads for developing pharmaceutical otoprotectants to alleviate an increasing clinical problem.


Diagnosis and root canal treatment in a mandibular premolar with three canals.
This paper presents a case report of a left mandibular second premolar with three canals and three different apical foramina. A 39-year-old male patient presented to our clinic with pain in the mandibular left second premolar. Initially, pain was caused by cold stimulus and later was spontaneously. The intraoral clinical examination revealed a fractured amalgam restoration with occlusal caries. Percussion and cold (Endo-Frost) tests were positive. The radiographic examination showed the presence of two roots. The probable diagnosis was an acute pulpitis. After access cavity, it was observed remaining roof of the pulp chamber and mild bleeding in the tooth lingual area, indicating the possible presence of a third canal. The endodontic treatment was completed in a single session using Root ZX apex locator and K3 NiTi rotary system with surgical diameter corresponding to a .02/45 file in the three canals and irrigation with 1% sodium hypochlorite. The canals were obtured with gutta-percha cones and Sealer 26 using the lateral condensation technique. After 1 year of follow-up, the tooth was asymptomatic and periapical repair was observed radiographically. Internal alterations should be considered during the endodontic treatment of mandibular second premolars. The correct diagnosis of these alterations by the analysis of preoperative radiographs can help the location of two or more canals, thereby avoiding root therapy failure.


Polyglutamine tracts regulate beclin 1-dependent autophagy.
Nine neurodegenerative diseases are caused by expanded polyglutamine (polyQ) tracts in different proteins, such as huntingtin in Huntington's disease and ataxin 3 in spinocerebellar ataxia type 3 (SCA3). Age at onset of disease decreases with increasing polyglutamine length in these proteins and the normal length also varies. PolyQ expansions drive pathogenesis in these diseases, as isolated polyQ tracts are toxic, and an N-terminal huntingtin fragment comprising exon 1, which occurs in vivo as a result of alternative splicing, causes toxicity. Although such mutant proteins are prone to aggregation, toxicity is also associated with soluble forms of the proteins. The function of the polyQ tracts in many normal cytoplasmic proteins is unclear. One such protein is the deubiquitinating enzyme ataxin 3 (refs 7, 8), which is widely expressed in the brain. Here we show that the polyQ domain enables wild-type ataxin 3 to interact with beclin 1, a key initiator of autophagy. This interaction allows the deubiquitinase activity of ataxin 3 to protect beclin 1 from proteasome-mediated degradation and thereby enables autophagy. Starvation-induced autophagy, which is regulated by beclin 1, was particularly inhibited in ataxin-3-depleted human cell lines and mouse primary neurons, and in vivo in mice. This activity of ataxin 3 and its polyQ-mediated interaction with beclin 1 was competed for by other soluble proteins with polyQ tracts in a length-dependent fashion. This competition resulted in impairment of starvation-induced autophagy in cells expressing mutant huntingtin exon 1, and this impairment was recapitulated in the brains of a mouse model of Huntington's disease and in cells from patients. A similar phenomenon was also seen with other polyQ disease proteins, including mutant ataxin 3 itself. Our data thus describe a specific function for a wild-type polyQ tract that is abrogated by a competing longer polyQ mutation in a disease protein, and identify a deleterious function of such mutations distinct from their propensity to aggregate.


[Laparoscopic fundoplication--what is proven?].
Laparoscopic fundoplication has become a popular procedure but several questions remain unresolved. Although randomised trials suggest that antireflux surgery is a good alternative to continuous medical therapy, the selection of patients for surgery remains crucial to achieve success. There is currently no proof that antireflux surgery will reduce the risk for developing esophageal adenocarcinoma. Complications, side effects and failures appear to be higher after a laparoscopic approach as compared to conventional open surgery, particularly in un-experienced hands. Partial fundoplications and tailored approaches have so far not shown convincingly better results than those that can be achieved by short and floppy 360 fundoplication.


Human immunodeficiency virus mutagenesis during antiviral therapy: impact of drug-resistant reverse transcriptase and nucleoside and nonnucleoside reverse transcriptase inhibitors on human immunodeficiency virus type 1 mutation frequencies.
The development of antiviral drug resistance is an important problem in the treatment of human immunodeficiency virus type 1 (HIV-1) infection. Potent antiretroviral therapy is currently used for treatment, and typically consists of at least two reverse transcriptase (RT) inhibitors. We have previously reported that both drugs and drug-resistant RT mutants can increase virus mutation frequencies. To further assess the contributions of nucleoside RT inhibitors (NRTIs), nonnucleoside RT inhibitors (NNRTIs), and drug-resistant RTs to HIV mutagenesis, a new high-throughput assay system was developed. This assay system was designed to specifically detect frameshift mutations in the luciferase gene in a single virus replication cycle. New drug-resistant RTs were identified that significantly altered virus mutation frequencies. Consistent with our previous observations of NRTIs, abacavir, stavudine, and zalcitabine increased HIV-1 mutation frequencies, supporting the general hypothesis that the NRTIs currently used in antiviral drug therapy increase virus mutation frequencies. Interestingly, similar observations were made with NNRTIs. This is the first report to show that NNRTIs can influence virus mutation frequencies. NNRTI combinations, NRTI-NNRTI combinations, and combinations of drug and drug-resistant RTs led to significant changes in the virus mutation frequencies compared to virus replication of drug-resistant virus in the absence of drug or wild-type virus in the presence of drug. This indicates that combinations of RT drugs or drugs and drug-resistant virus created during the evolution of drug resistance can act together to increase HIV-1 mutation frequencies, which would have important implications for drug therapy regimens. Finally, the influence of drug-resistant RT mutants from CRF01_AE viruses on HIV-1 mutation frequencies was analyzed and it was found that only a highly drug resistant RT led to altered virus mutation frequencies. The results further suggest that high-level drug-resistant RT can significantly influence virus mutation frequencies. A structural model that explains the mutation frequency data is discussed.


Trend in proportions of missed children during polio supplementary immunization activities in the African Region: evidence from independent monitoring data 2010-2012.
This is a comparative analysis of independent monitoring data collected between 2010 and 2012, following the implementation of supplementary immunization activities (SIAs) in countries in the three sub regional blocs of World Health Organization in the African Region. The sub regional blocs are Central Africa, West Africa, East and Southern Africa. In addition to the support for SIAs, the Central and West African blocs, threatened with importation and re-establishment of polio transmission received intensive coordination through weekly teleconferences. The later, East and Southern African bloc with low polio threats was not engaged in the intensive coordination through teleconferences. The key indicator of the success of SIAs is the proportion of children missed during SIAs. The results showed that generally there was a decrease in the proportion of children missed during SIAs in the region, from 7.94% in 2010 to 5.95% in 2012. However, the decrease was mainly in the Central and West African blocs. The East and Southern African bloc had countries with as much as 25% missed children. In West Africa and Central Africa, where more coordinated SIAs were conducted, there were progressive and consistent drops, from close to 20-10% at the maximum. At the country and local levels, steps were undertaken to ameliorate situation of low immunization uptake. Wherever an area is observed to have low coverage, local investigations were conducted to understand reasons for low coverage, plans to improve coverage are made and implemented in a coordinated manner. Lessons learned from close monitoring of polio eradication SIAs are will be applied to other campaigns being conducted in the African Region to accelerate control of other vaccine preventable diseases including cerebrospinal meningitis A, measles and yellow fever.


Chemotaxis as an emergent property of a swarm.
We have characterized and quantified a form of bacterial chemotaxis that manifests only as an emergent property by measuring symmetry breaking in a swarm of Myxococcus xanthus exposed to a two-dimensional nutrient gradient from within an agar substrate. M. xanthus chemotaxis requires cell-cell contact and coordinated motility, as individual motile cells exhibit only nonvectorial movement in the presence of a nutrient gradient. Genes that specifically affect M. xanthus chemotaxis include at least 10 of the 53 that express enhancer binding proteins of the NtrC-like class, an indication that this behavior is controlled through transcription, most likely by a complex signal transduction network.


Staudinger/aza-Wittig reaction to access Nβ-protected amino alkyl isothiocyanates.
A unified approach to access Nβ-protected amino alkyl isothiocyanates using Nβ-protected amino alkyl azides through a general strategy of Staudinger/aza-Wittig reaction is described. The type of protocol used to access isothiocyanates depends on the availability of precursors and also, especially in the amino acid chemistry, on the behavior of other labile groups towards the reagents used in the protocols; fortunately, we were not concerned about both these factors as precursor-azides were prepared easily by standard protocols, and the present protocol can pave the way for accessing title compounds without affecting Boc, Cbz and Fmoc protecting groups, and benzyl and tertiary butyl groups in the side chains. The present strategy eliminates the need for the use of amines to obtain title compounds and thus, this method is step-economical; additional advantages include retention of chirality, convenient handling and easy purification. A few hitherto unreported compounds were also prepared, and all final compounds were completely characterized by IR, mass, optical rotation, and 1H and 13C NMR studies.


[Autoimmunity and Freiburg classification in common variable immunodeficiency].
Up to 25% of patients with common variable immunodeficiency (CVID) debut with autoimmunity, which is related to the Freiburg classification, which is based on flow cytometry. to determine the frequency and type of autoimmune diseases and their association with the Freiburg classification in adults with CVID. A cross-sectional, analytical and observational study was carried out with 33 patients belonging to the Primary Immunodeficiency Clinic of a third level hospital, with a diagnosis of CVID. They were divided into 3 phenotypes according to the Freiburg classification. Of the 33 patients studied, 66.6% presented autoimmune diseases, 19 of them (86.3%) had cytopenia; 42.1% belonged to Freiburg group Ia, 36.8% to Ib and 21% to phenotype II. In 36.6% of the patients, autoimmune cytopenia were the first manifestation of CVID; and up to 70% of them belong to the Freiburg phenotype Ia (p = 0.086). Patients with autoimmune cytopenia had a lower percentage of isotype-switched memory B cells (p = 0.018), no higher percentage of CD21low B cells (p = 0.226). Classification by CVID phenotypes allows the identification of the patient's profile according to the percentage of memory B cells with isotype change, which is useful to intentionally search for non-infectious complications of the disease.


Evidence for presence of proinsulin-immunoreactive glycoprotein(s) in fetal bovine pancreatic extracts.
A large-molecular-weight proinsulin-immunoreactive protein fraction was obtained from an extract of fetal bovine pancreases by gel filtration in 6 M guanidine-1 M acetic acid. Concanavalin A-Sepharose-affinity column chromatography of the large-molecular-weight fraction yielded a discrete alpha-methyl-mannoside-displaceable immunoreactive peak that also displayed N-acetylglucosamine-specific binding to wheat germ lectin-Sepharose. Chemically tritiated and radioiodinated lectin-reactive proteins interacted specifically with antibodies to insulin and bovine proinsulin. Immunochemically purified (reaction with antibodies followed by separation of antigen-antibody complexes on protein A-Sepharose) radiolabeled lectin-reactive proteins were analyzed by gel filtration in guanidine-acetic acid and by sodium dodecyl sulfate polyacrylamide gel electrophoresis after disulfide bond-cleavage treatments. Results from these studies suggest the existence of an approximately 67,000-Mr glycoprotein that contains antigenic domains common to proinsulin and insulin.


Staff sizing in the material and sterilization center of a university hospital.
To apply the standard time parameters of nursing activities proposed by COFEN Resolution No. 543/2017 to determine the number of nursing technicians in the material and sterilization center of a university hospital and compare the projected framework with the existing one in the sector. An exploratory and descriptive study with data collection performed on the sterilization control sheets and direct observation of the work process stages in the different sector areas. Only one of the 15 described activities was not conducted in the sector; those most performed which presented more workload and required more personnel were the receiving of contaminated materials and the organization and distribution of sterilized materials; the areas that required the highest number of employees were for materials preparation and cleaning/disinfection. No difference was observed between the number of staff required in the different work shifts, and the calculated staff quantity was similar to that existing in the result of the staff sizing performed using the functional sites. The proposals analyzed in this study can be used to calculate staff size in the studied sector and in other institutions.