Reduced miR-202 levels enhanced oral cancer development via targeting Sp1.
The current study aimed to evaluate the possible role of microRNA (miR)-202 in the development of oral cancer. First, miR-202 levels were found to be decreased in the serum and tissues of oral cancer patients compared with healthy controls. Receiver operating characteristic analysis was carried out to explore the diagnostic value of serum miR-202 for oral cancer. Overexpression of miR-202 significantly decreased the migratory capacity of SCC-9 cells, while inhibition of miR-202 markedly increased the migratory capacity of SCC-9 cells. Moreover, the invasive capacity was decreased in SCC-9 cells transfected with an miR-202 mimic. In addition, the invasive capacity was enhanced in SCC-9 cells transfected with an miR-202 inhibitor. A dual luciferase reporter assay showed that overexpression of miR-202 markedly suppressed the relative luciferase activity of the pmirGLO-SP1-3'untranslated region. Overexpression of miR-202 suppressed the protein level of Sp1, but inhibition of miR-202 markedly enhanced the protein expression of Sp1. Inhibition of miR-202 enhanced the phosphorylation of protein kinase B. Additionally, the correlations between the expression levels of Sp1 and miR-202 and the clinicopathological factors of oral cancer were analyzed. The results showed that patients with high expression of Sp1 and miR-202 progressed to earlier clinical stages, had deeper infiltration depths and were more prone to lymph node metastasis compared with the healthy controls. In conclusion, the current study presented novel data indicating that decreased miR-202 enhanced the progression of oral cancer via Sp1.


Glucagon antagonists. Synthesis and inhibitory properties of Asp3-containing glucagon analogs.
In an effort to find analogs of glucagon that would bind to the glucagon receptor of the rat liver membrane but would not activate membrane-bound adenyl cyclase, several hybrid molecules were synthesized which contained sequences from both glucagon and secretin. [Asp3, Glu9]Glucagon and [Asp3, Glu9, Arg12]glucagon were inactive in the adenyl cyclase assay even at high concentrations but retained some binding affinity for the receptor. They were able to displace 125I-glucagon completely from its receptor and could completely inhibit the activation of adenyl cyclase by natural or synthetic glucagon. The inhibition index [I/A]50 was approximately 110 for both analogs. [Asp3]Glucagon, [Glu3]glucagon and [Asp3, Lys17, 18, Glu21]glucagon were weak partial agonists, while [Asp3, Glu21]glucagon was inactive and a poor inhibitor. The peptides were synthesized by solid-phase methods and purified to homogeneity by reverse-phase high-performance liquid chromatography on C18 silica columns. These are the first fully synthetic competitive glucagon antagonists to be reported.


Quasiclassical trajectory study of the SiH(4)+H-->SiH(3)+H(2) reaction on a global ab initio potential energy surface.
The SiH(4)+H-->SiH(3)+H(2) reaction has been investigated by the quasiclassical trajectory (QCT) method on a recent global ab initio potential energy surface [M. Wang et al., J. Chem. Phys. 124, 234311 (2006)]. The integral cross section as a function of collision energy and thermal rate coefficient for the temperature range of 300-1600 K have been obtained. At the collision energy of 9.41 kcalmol, product energy distributions and rovibrational populations are explored in detail, and H(2) rotational state distributions show a clear evidence of two reaction mechanisms. One is the conventional rebound mechanism and the other is the stripping mechanism similar to what has recently been found in the reaction of CD(4)+H [J. P. Camden et al., J. Am. Chem. Soc. 127, 11898 (2005)]. The computed rate coefficients with the zero-point energy correction are in good agreement with the available experimental data.


Initial immune response after exposure to Mycobacterium tuberculosis or to SARS-COV-2: similarities and differences.
Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb) and Coronavirus disease-2019 (COVID-19), whose etiologic agent is severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), are currently the two deadliest infectious diseases in humans, which together have caused about more than 11 million deaths worldwide in the past 3 years. TB and COVID-19 share several aspects including the droplet- and aerosol-borne transmissibility, the lungs as primary target, some symptoms, and diagnostic tools. However, these two infectious diseases differ in other aspects as their incubation period, immune cells involved, persistence and the immunopathological response. In this review, we highlight the similarities and differences between TB and COVID-19 focusing on the innate and adaptive immune response induced after the exposure to Mtb and SARS-CoV-2 and the pathological pathways linking the two infections. Moreover, we provide a brief overview of the immune response in case of TB-COVID-19 co-infection highlighting the similarities and differences of each individual infection. A comprehensive understanding of the immune response involved in TB and COVID-19 is of utmost importance for the design of effective therapeutic strategies and vaccines for both diseases.


Isolation of cytotoxic diterpenoids from the Chilean medicinal plant Azorella compacta Phil from the Atacama Desert by high-speed counter-current chromatography.
Azorella compacta (Apiaceae) is a native Chilean cushion shrub which produces a resin containing mulinane and azorellane diterpenoids. This plant has been used since pre-Colombian times to treat inflammation and dental neuralgias. In this work the first preparative fractionation of diterpenoids present in this plant by means of high-speed counter-current chromatography (HSCCC) was applied, and cytotoxic effects of the isolated compounds were evaluated for the first time against a panel of MCF7 cells. The major compounds isolated were identified by means of spectroscopy as azorellanol, 13α, 14α-dihydroxymulin-11-en-20-oic acid, mulinolic acid, mulin-11,13-dien-20-oic acid, 17-acetoxy-mulin-11,13-dien-20 oic acid, and 17-acetoxy-mulinic acid (compounds 7, 9-11 and 13, respectively), and four minor diterpenoids [7-deacetyl-azorellanol (6), 13-epi-azorellanol, 7-acetoxy-mulin-9,12-diene, and 17-acetoxy-mulin-11,13-dien-20-oic acid (compounds 4, 8 and 12)], together with three new minor diterpenoids: 13β,14β-dihydroxymulin-11-en-20-oic acid (1), 13-epiazorellanone (2) and 13-epi-7-deacetyl-azorellanol (3) were identified. Besides, compounds 4, 6, 7, 8 and 11 displayed good cytotoxic activity (less than 50% cell viability at 100 µM). Among them, compound 7, an acetylated azorellane, was the most active. HSCCC allowed the isolation of 13 diterpenoids present in A. compacta. Three compounds are reported for the first time. Isolated azorellanes are more potent cytotoxic agents than are mulinanes. © 2015 Society of Chemical Industry.


Transition Coverage and Clarity in Self-Insured Corporate Health Insurance Benefit Plans.
Purpose: While many health insurance plans now cover at least some gender-affirming care for transgender persons, no study to date has examined contract language about gender-affirming care in self-insured corporate plans. We sought to evaluate private company offerings from the perspective of an employee, analyzing clarity as well as what gender-affirming care is covered or excluded. Methods: We coded 435 health insurance contracts from 40 U.S. self-insured corporations from 2019 for inclusion of 52 coverage aspects from the World Professional Association for Transgender Health (WPATH) recommendations. We categorize contracts by clarity of the document and its inclusions and exclusions, and compare each company's contract ratings to their 2019 Human Rights Campaign (HRC) Workplace Equality Index rating. Results: Findings reveal higher levels of total exclusions in contracts (9% here vs. 3% found in prior studies of more highly regulated plans), as well as extensive variation in clarity, coverage specifications, and types of exclusions. Facial confirmation surgery procedures are commonly excluded even in plans that affirm the WPATH guidelines. Twenty-five percent of the companies in the study offered at least one contract with a categorical exclusion. HRC ratings did not match up to our ratings of gender-affirming coverage. Conclusion: Legal complexity has resulted in a patchwork of continued health insurance exclusions of gender-affirming care even as coverage has expanded. Lack of transparency and clarity also contributes to challenges in understanding one's own coverage as well as mapping the national picture of transgender inclusion in health care plans.


Targeting BCL-2-like Proteins to Kill Cancer Cells.
Mutations that impair apoptosis contribute to cancer development and reduce the effectiveness of conventional anti-cancer therapies. These insights and understanding of how the B cell lymphoma (BCL)-2 protein family governs apoptosis have galvanized the search for a new class of cancer drugs that target its pro-survival members by mimicking their natural antagonists, the BCL-2 homology (BH)3-only proteins. Successful initial clinical trials of the BH3 mimetic venetoclax/ABT-199, specific for BCL-2, have led to its recent licensing for refractory chronic lymphocytic leukemia and to multiple ongoing trials for other malignancies. Moreover, preclinical studies herald the potential of emerging BH3 mimetics targeting other BCL-2 pro-survival members, particularly myeloid cell leukemia (MCL)-1, for multiple cancer types. Thus, BH3 mimetics seem destined to become powerful new weapons in the arsenal against cancer. This review sketches the discovery of the BCL-2 family and its impact on cancer development and therapy; describes how interactions of family members trigger apoptosis; outlines the development of BH3 mimetic drugs; and discusses their potential to advance cancer therapy.


Altered cellular responses by varying expression of a ribosomal protein gene: sequential coordination of enhancement and suppression of ribosomal protein S3a gene expression induces apoptosis.
A growing body of evidence indicates that individual ribosomal proteins and changes in their expression, participate in, and modulate, a variety of cellular activities. Our earlier studies have found that apoptosis could be induced by inhibiting expression of ribosomal protein S3a (RPS3a) in many tumor cells which constitutively express RPS3a at levels much higher than in normal cells. This study aimed to investigate cellular responses to enhancement of RPS3a expression, and whether apoptosis could be induced by sequential alterations in RPS3a expression involving enhancement from an initially low constitutive level, followed by suppression. Stably transfected NIH 3T3- derived cell lines were established in which exogenous RPS3a expression could be readily manipulated. Enhancement of RPS3a expression appeared to induce transformation as assessed by well-established criteria such as foci formation and anchorage-independent growth in vitro, and formation of tumors in nude mice. These properties were compared with those observed in ras-transformed NIH 3T3 cells. Apparent transformation occurred only when enhanced RPS3a-expressing cells were in close cell-cell contact. Suppression of enhanced RPS3a expression was observed to induce apoptosis as assessed by various morphological and biochemical characteristics including cell shrinkage, membrane blebbing, chromatin condensation, nuclear and cell fragmentation, phosphatidylserine externalization, and internucleosomal DNA fragmentation. This induction of apoptosis was not specific to apparently transformed cells, as cells at low confluence, which likewise expressed RPS3a at enhanced levels but exhibited no morphological transformation, underwent apoptosis when RPS3a expression was inhibited. These results support a role for RPS3a in the apoptotic process, but not as an oncoprotein per se.


Biological X-ray irradiator characterization for use with small animals and cells.
This study presents the characterization of an X-ray irradiator through dosimetric tests, which confirms the actual dose rate that small animals and cells will be exposed to during radiobiological experiments. We evaluated the linearity, consistency, repeatability, and dose distribution in the positions in which the animals or cells are placed during irradiation. In addition, we evaluated the performance of the X-ray tube (voltage and tube operating current), the radiometric survey (leakage radiation) and safety devices. The irradiator default setting was established as 160 kV and 25 mA. Tests showed that the dose rate was linear overtime (R2=1) and remained stable for long (constant) and short (repeatability) intervals between readings. The mean dose rate inside the animal cages was 1.27±0.06 Gy/min with a uniform beam of 95.40% (above the minimum threshold guaranteed by the manufacturer). The mean dose rate inside the cell plates was 0.92±0.19 Gy/min. The dose rate dependence with tube voltage and current presented a quadratic and linear relationship, respectively. There was no observed mechanical failure during evaluation of the irradiator safety devices and the radiometric survey obtained a maximum ambient equivalent dose rate of 0.26 mSv/h, which exempts it from the radiological protection requirements of the International Atomic Energy Agency. The irradiator characterization enables us to perform radiobiological experiments, and assists or even replaces traditional therapy equipment (e.g., linear accelerators) for cells and small animal irradiation, especially in early research stages.


Declining dental caries in Jerusalem schoolchildren.
There has been an increase of dental caries in Israel during the last decade (1970-80), while most developed industrialized countries reported a decline in dental caries since 1970. A survey carried out on 267 first graders (6 yr old) by the same investigator and with the same methods as in the previous survey (1977), showed a significant decrease in prevalence of dental caries in 1985 as follows: 78% affected children with a caries score of 4.9 compared to 86.2% with a caries score of 5.2 in 1977.


Correlation of COX- 2 Expression in Colorectal Carcinoma with Clinicopathological Features.
Colorectal carcinoma is the most common neoplasm of the gastrointestinal tract. COX-2 expression is upregulated in colorectal carcinoma. Therefore its assessment would identify patients amenable to adjuvant anticyclooxygenase therapy. We studied COX-2 immunoexpression in colorectal adenocarcinoma and correlated it with clinicopathological features as age, gender, tumor location, tumor size, degree of differentiation, and TNM stage. Sixty-five consecutive cases of colorectal adenocarcinoma were retrieved from the records of the Pathology Department of a tertiary care hospital. The tumors were categorized as low positive and high positive based on their total COX-2 scores, which is the sum of proportion and intensity scores of immunostaining, and were correlated with clinicopathological features. Statistical analysis was done using the Chi-square test and Kendall's Tau-b correlation method. COX-2 was expressed in 86.2% of cases including 90% of left colonic carcinomas and 77.3% of right colonic carcinomas. Lymph node metastasis was present in 22.2%, 25% and 47.75% of COX-2 negative, low positive and high positive tumors respectively. High positive COX-2 cases constituted 33.3% of stage I, 58.8% of stage II, 80% of stage III and 100% of stage IV tumors. About 56.6% of well differentiated, 66.6% of moderately differentiated and 100% of poorly differentiated carcinomas showed high COX-2 expression. The COX-2 overexpression was associated with advancing depth of invasion (p=0.021), stage of tumor (p=0.05), more frequent lymph node metastasis and decreasing degree of differentiation. The association of COX-2 overexpression with increasing stage and depth of invasion may justify the use of COX-2 inhibitors as an adjuvant to chemo and radiotherapy.


Active-State Structures of a Small Heat-Shock Protein Revealed a Molecular Switch for Chaperone Function.
Small heat-shock proteins (sHsps) maintain cellular homeostasis by binding to denatured client proteins to prevent aggregation. Numerous studies indicate that the N-terminal domain (NTD) of sHsps is responsible for binding to client proteins, but the binding mechanism and chaperone activity regulation remain elusive. Here, we report the crystal structures of the wild-type and mutants of an sHsp from Sulfolobus solfataricus representing the inactive and active state of this protein, respectively. All three structures reveal well-defined NTD, but their conformations are remarkably different. The mutant NTDs show disrupted helices presenting a reformed hydrophobic surface compatible with recognizing client proteins. Our functional data show that mutating key hydrophobic residues in this region drastically altered the chaperone activity of this sHsp. These data suggest a new model in which a molecular switch located in NTD facilitates conformational changes for client protein binding.


Biological activity of toxic shock syndrome toxin 1 and a site-directed mutant, H135A, in a lipopolysaccharide-potentiated mouse lethality model.
A recombinant of toxic shock syndrome toxin 1 (TSST-1) which contains a single histidine-to-alanine mutation at residue 135 (H135A) was analyzed for toxicity and vaccine potential in a lipopolysaccharide (LPS)-potentiated mouse lethality model. The 50% lethal dose (LD50) of TSST-1 in BALB/c mice was 47.2 micrograms/kg, but H135A was not lethal when tested at a dose equivalent to 10 LD50s of TSST-1. Levels of tumor necrosis factor (TNF) and gamma interferon (IFN-gamma) in serum were, respectively, 10- and 50-fold higher in LPS-potentiated mice injected with 15 LD50s of TSST-1 than in mice given H135A. Mice injected with only TSST-1 did not have elevated levels of TNF or IFN-gamma in serum, while H135A plus LPS or LPS alone elicited identical, yet very low, levels of TNF and IFN-gamma. An enzyme-linked immunosorbent assay of H135A and TSST-1 with anti-TSST-1 serum yielded very similar dose-response curves, which strongly suggests that H135A serologically and conformationally resembles the native toxin. Mice immunized with H135A developed antibodies that recognized TSST-1 in an enzyme-linked immunosorbent assay and afforded protection against a 15-LD50 challenge of TSST-1 plus LPS. The pooled sera of mice immunized with either TSST-1 or H135A also prevented lymphocyte proliferation due to TSST-1.


Physicians in transition: practice due diligence.
The landscape of healthcare is changing rapidly. That landscape is now a business model of medicine. That rapid change resulting in a business model is affecting physicians professionally and personally. The new business model of medicine has led to large healthcare organizations hiring physicians as employees. The role of a physician as an employee has many limitations in terms of practice and personal autonomy. Employed physicians sign legally binding employment agreements that are written by the legal team working for the healthcare organization. Thus physicians should practice due diligence before signing the employment agreement. "Due diligence" refers to the care a reasonable person should take before entering into an agreement with another party. That reasonable person should seek expertise to represent his or her interests when searching a balanced agreement between the physician and organization.


Clinical Features and Imaging Findings of Myelin Oligodendrocyte Glycoprotein-IgG-Associated Disorder (MOGAD).
Myelin oligodendrocyte glycoprotein-IgG-associated disorder (MOGAD) is a nervous system (NS) demyelination disease and a newly recognized distinct disease complicated with various diseases or symptoms; however, MOGAD was once considered a subset of neuromyelitis optica spectrum disorder (NMOSD). The detection of MOG-IgG has been greatly improved by the cell-based assay test method. In one study, 31% of NMOSD patients with negative aquaporin-4 (AQP-4) antibody were MOG-IgG positive. MOGAD occurs in approximately the fourth decade of a person's life without a markedly female predominance. Usually, optic neuritis (ON), myelitis or acute disseminated encephalomyelitis (ADEM) encephalitis are the typical symptoms of MOGAD. MOG-IgG have been found in patients with peripheral neuropathy, teratoma, COVID-19 pneumonia, etc. Some studies have revealed the presence of brainstem lesions, encephalopathy or cortical encephalitis. Attention should be given to screening patients with atypical symptoms. Compared to NMOSD, MOGAD generally responds well to immunotherapy and has a good functional prognosis. Approximately 44-83% of patients undergo relapsing episodes within 8 months, which mostly involve the optic nerve, and persistently observed MOG-IgG and severe clinical performance may indicate a polyphasic course of illness. Currently, there is a lack of clinical randomized controlled trials on the treatment and prognosis of MOGAD. The purpose of this review is to discuss the clinical manifestations, imaging features, outcomes and prognosis of MOGAD.


Medical ethics: a brief response to Seedhouse.
Medical ethics is that branch of applied philosophy which considers issues of values raised by medical practice, and should not be equated with 'principlism'. Clarification of facts/values distinctions is an important part of this work. The notion that medical philosophy can flourish in the hands of medical 'generalists' without specialist philosophers, is misguided. Both must work together to promote right reason and right action in medical education and practice.


Cellular mechanosensing of the biophysical microenvironment: A review of mathematical models of biophysical regulation of cell responses.
Cells in vivo reside within complex microenvironments composed of both biochemical and biophysical cues. The dynamic feedback between cells and their microenvironments hinges upon biophysical cues that regulate critical cellular behaviors. Understanding this regulation from sensing to reaction to feedback is therefore critical, and a large effort is afoot to identify and mathematically model the fundamental mechanobiological mechanisms underlying this regulation. This review provides a critical perspective on recent progress in mathematical models for the responses of cells to the biophysical cues in their microenvironments, including dynamic strain, osmotic shock, fluid shear stress, mechanical force, matrix rigidity, porosity, and matrix shape. The review highlights key successes and failings of existing models, and discusses future opportunities and challenges in the field.


Use of serum protein concentration to predict mortality in mixed-source dairy replacement heifers.
The relationship between serum protein concentration in the 1st week of life and survival to 16 weeks of age was examined in 3,479 Holstein replacement heifers over a period of 10 years on a farm with endemic salmonellosis. Thirty-four percent of calves studied had serum protein concentrations < 5.0 g/dL and 60.5% of calves had serum protein concentrations < 5.5 g/dL. Cumulative mortality was 7.9%, indicating that calves with marginal passive transfer status can be reared successfully under conditions of endemic salmonellosis. Optimal survival was observed in calves with serum protein concentrations > 5.5 g/dL. Calves with serum protein concentrations of 5.0-5.4 g/dL had only a slightly increased relative risk (RR) of mortality (RR = 1.3) compared to calves with serum protein concentrations > 5.5 g/dL. The highest RR was experienced by calves with serum protein concentrations < 4 g/dL (RR = 4.6) and 4.0-4.4 g/dL (RR = 3.1). Calves with inadequate passive transfer (serum protein concentration < 5.0 g/dL), experienced increased mortality until at least 10 weeks of age, indicating that failure of passive transfer has an effect on calf health that extends into the juvenile period. Models in which serum protein concentration was treated with as a continuous variable or as a categorical variable failed to demonstrate any significant interaction between baseline mortality and the RR of mortality. This finding suggests that the RR derived in the present study should be applicable to farms with dramatically different baseline mortality rates.


Sleep impairment in patients with painful diabetic peripheral neuropathy.
This study evaluated sleep impairment associated with painful diabetic peripheral neuropathy (DPN), a neuropathic pain condition. Sleep is of critical concern for DPN because sleep impairment and its comorbidities may influence type 2 diabetes progression. This is a supplemental analysis of sleep data from a burden of illness study of patients with painful DPN (N=255, 61+/-12.8 y old, 51.4% women). Sleep was evaluated using the Medical Outcomes Study Sleep measure (MOS-Sleep). MOS-Sleep scores were compared with general population norms (N=1011), the MOS chronic disease sample (N=3445), and patients with postherpetic neuralgia (N=89). The MOS-Sleep Sleep Adequacy score was compared with data from the MOS diabetes subsample (N=590). Patients with painful DPN reported impaired sleep relative to the general population (P<0.001), the chronic disease sample (P<0.001), and postherpetic neuralgia patients (P<0.05). Self-rated MOS-Sleep Sleep Adequacy was significantly less for the painful DPN than for the diabetes sample (P<0.001), although self-reported hours of sleep were not significantly different. Multiple regression indicated that age, average daily pain, and anxiety and depression symptom levels were each significantly (P<0.01) associated with, and collectively accounted for, 47% of variance in the MOS-Sleep Sleep Problems Index. Painful DPN is associated with considerable sleep impairment. Given the recognized association between sleep impairment, type 2 diabetes and metabolic and affective disturbance, and the known adverse impact of affective disturbance on diabetes self-care, addressing these features-pain, sleep, and affective disturbance-is an important aspect of care for patients with painful DPN.


A hybrid group-based movie recommendation framework with overlapping memberships.
Recommender Systems (RS) are widely used to help people or group of people in finding their required information amid the issue of ever-growing information overload. The existing group recommender approaches consider users to be part of a single group only, but in real life a user may be associated with multiple groups having conflicting preferences. For instance, a person may have different preferences in watching movies with friends than with family. In this paper, we address this problem by proposing a Hybrid Two-phase Group Recommender Framework (HTGF) that takes into consideration the possibility of users having simultaneous membership of multiple groups. Unlike the existing group recommender systems that use traditional methods like K-Means, Pearson correlation, and cosine similarity to form groups, we use Fuzzy C-means clustering which assigns a degree of membership to each user for each group, and then Pearson similarity is used to form groups. We demonstrate the usefulness of our proposed framework using a movies data set. The experiments were conducted on MovieLens 1M dataset where we used Neural Collaborative Filtering to recommend Top-k movies to each group. The results demonstrate that our proposed framework outperforms the traditional approaches when compared in terms of group satisfaction parameters, as well as the conventional metrics of precision, recall, and F-measure.


Calciate-mediated intermolecular hydroamination of diphenylbutadiyne with secondary anilines.
Calciate-mediated intermolecular hydroamination of diphenylbutadiyne with N-phenyl and N-isopropyl-substituted anilines yields E- and Z-isomers of the corresponding 1-anilino-1,4-diphenylbut-1-ene-3-yne. In the case of HNPh(2) solely heterobimetallic K(2)Ca(NPh(2))(4) is able to effectively catalyze this hydroamination reaction in tetrahydrofuran at elevated temperatures.


Loss of K+ATP-channel-mediated vasodilation after induction of tachyphylaxis to peroxynitrite.
Systemic injections of peroxynitrite elicit pronounced vasodilator responses in rats by activation of ATP-dependent K+ channels (K+ATP-channels). The aim of this study was to determine whether development of tachyphylaxis to the vasodilator actions of peroxynitrite involves the loss of K+ATP-channel function. The falls in mean arterial blood pressure (MAP) and mesenteric and hindquarter vascular resistances produced by the K+ATP-channel agonist, cromakalim (3-18 microg/kg, iv), and the nitric oxide (NO) donor, sodium nitroprusside (SNP; 1-4 microg/kg, iv), were determined in pentobarbital-anesthetized rats before and after induction of tachyphylaxis to peroxynitrite induced by the administration of 10 injections of peroxynitrite (10 micromol/kg, iv). The first dose of peroxynitrite elicited pronounced falls in MAP and vascular resistances whereas the tenth injection elicited much smaller responses that were equivalent to those of decomposed peroxynitrite. Before induction of tachyphylaxis to peroxynitrite, cromakalim and SNP produced dose-dependent reductions in MAP and vascular resistances. The hemodynamic actions of cromakalim were markedly attenuated after induction of tachyphylaxis to peroxynitrite whereas the SNP-induced responses were only slightly attenuated. These results suggest that tachyphylaxis to the vasodilator actions of peroxynitrite involves the loss of K+ATP-channel function whereas tachyphylaxis to peroxynitrite minimally affects NO-mediated vasodilation. Taken together, these findings raise the possibility that peroxynitrite inhibits K+ATP-channel function by oxidation and/or nitration of amino acids in these channels.


Detection of Chronic Kidney Disease by Using Different Equations of Glomerular Filtration Rate in Patients with Type 2 Diabetes Mellitus: A Cross-Sectional Analysis.
Introduction Chronic kidney disease (CKD) is a global threat due to its high mortality. It is essential to know the actual magnitude of diabetic CKD to design a specific management program. However, there is limited knowledge regarding the most suitable equation to measure CKD in patients with Type 2 diabetes mellitus (T2DM). This paper aimed to analyze estimated glomerular filtration rate (eGFR) based on different equations to detect the CKD among T2DM. Methods A hospital-based cross-sectional study was carried out, and a clinical registry was used to collect 4,042 T2DM patients from a large district hospital in Northeast Thailand. CKD patients were diagnosed when eGFR was less than 60 ml/min/1.73m2. Using Stata statistical software (StataCorp LP, College Station, TX), three standard equations, such as 'modification of diet in renal disease (MDRD-4)', 'chronic kidney disease epidemiology collaboration (CKD-EPI)', and 'Cockcroft-Gault (C-G)' equations, were used to produce eGFR values to report and compare stages of CKD. Results The mean age of the patients was 61.4 (± 10.7) years and male to female ratio was 1:1.9. According to the MDRD-4, CKD-EPI, and C-G equation, the prevalence of diabetic CKD was 21.4%, 21.9%, and 31.4%, respectively, and the frequency of CKD Stage 3 to 5 was found to be different among T2DM. About 3,789 (93.9%) measurements appeared to be classified as different stages of CKD (Stages 1 to 5) between MDRD-4 and CKD-EPI equations (kappa: 0.905; 95% confidence interval (CI): 0.83 - 0.97, p < 0.001). However, this study found that the above-mentioned agreement was 70.9% between CKD-EPI and C-G equation (kappa: 0.56, 95% CI: 0.44 - 0.67, p < 0.001). Conclusions CKD-EPI equations can overcome the constraint of MDRD-4 and C-G equations to report CKD and can be used in patients with T2DM.


Impact of Exercise on Fatigue in Patients Undergoing Dialysis in a Tertiary Care Hospital.
Background Chronic kidney disease (CKD) is a current public health problem associated with progression to end-stage renal disease (ESRD), cardiovascular disease, and increased mortality rates. The disease is progressive. It is estimated that there are about 20-25 patients with milder kidney damage for every patient on renal replacement therapy. Physical activity is one of the critical elements for the prevention of chronic diseases and exercises in CKD help to improve muscular strength, cardiorespiratory endurance, overall function, and quality of life. Fatigue can cause an inability to perform exercises and can affect physiological and psychological function. There is a need to analyze the effects of exercises on fatigue in outpatients undergoing dialysis in the Indian rural population. Methods This study was a randomized, controlled, interventional, single-center trial. The participants diagnosed with ESRD and who were on hemodialysis were randomly assigned to group A, the exercise group (EG), which had exercise training during dialysis, and group B, the control group (CG), which had no added exercises during dialysis or at home and followed a standard routine method. The outcome measure of fatigue was evaluated through the total Fatigue Assessment Scale (FAS) score at 0, 12, 24, and 36 weeks. The exercise was pilot tested and planned based on the guidelines and carried out during hemodialysis two days a week. It was followed up with a five-day home exercise program when the patients did not undergo dialysis. Results A total of 48 participants were chosen for the study, of which 30 participants completed 36 weeks of training, with exercise adherence of more than 60%. There was a statistically significant difference in FAS between the two groups (F (3, 84) = 10.513, P < 0.001) at a 95% confidence interval (P < 0.05). Post hoc comparisons between groups at baseline and at 12 weeks indicated that there was no significant difference in FAS (P = 0.271 and P = 0.08), but recorded a significant difference (P = 0.001) at 24 and 36 weeks, respectively, between the EG and CG. Conclusions The results indicate that the intradialytic exercise intervention was effective in reducing the level of fatigue in outpatients undergoing dialysis on a long-term exercise program.


Efficient, fast and inexpensive bioassay to monitor benthic microalgae toxicity: Application to Ostreopsis species.
Even though HPLC-MS is commonly used to quantify the toxin content of Ostreopsis spp. cells, there is a need to develop easy-to-use toxicological tests to set thresholds during Ostreopsis spp. blooms. The crustacean Artemia has been widely used to evaluate the presence and toxicity of chemicals and biological contaminants and we anticipated that it could also be useful to test Ostreopsis spp. toxicity. Its relevance was first assessed by investigating the variability of the toxic effects among Ostreopsis spp. strains and throughout the dinoflagellate life cycle in combination with chemical analyses of the toxinic content by UHPLC-HRMS. After testing the toxicity of fractions prepared from Ostreopsis spp. cells, the known ova- and paly-toxins were not the only toxic metabolites to Artemia franciscana, indicating that other toxic compounds synthesized by Ostreopsis spp. still remain to be identified. To extend the bioassay to in situ monitoring, the toxicity of the benthic microalgal consortium was tested during a natural bloom of Ostreopsis cf. ovata in the NW Mediterranean Sea. The results highlight the accuracy and sensitivity of the ecotoxicological assay with Artemia franciscana to assess the toxicity of Ostreopsis spp. blooms.


Aging in Tunisia.
Owing in part to its diversity of citizenship, Tunisia has a rich history of economic and social changes-including some of the same age-based demographic changes witnessed globally. Faced with an increasing older population and fewer young adults available to provide support, the government of Tunisia has implemented policies for the older population. These new governmental policies are intended to provide limited funds for older adults to supplement financial assistance provided by their families. Yet, even with a demographic transition occurring, recent published research on the aging Tunisian adult is limited at best. Policy change and research that improve the well-being of Tunisia's aging population must be based on adequate data. Thus, understanding the state of aging in Tunisia provides policymakers and researchers more complete information to use in decision making. Further studies should address more aspects of the Tunisian aging experience and add data that are more contemporary.


[Effects of extraction methods on soil water isotope and plant water source segmentation.]
We used two types of soil with different physicochemical properties (loam and sand), oven-dried them, and then added the known isotopic composition mineral water that was reference water to compose the soil-water mixture with different soil water contents (loam: 0.15, 0.20, 0.30 g·g-1; sand: 0.10 g·g-1). After that, we set up different equilibrium time (loam: 3, 6, 12, 24, 48, 72, 96 h; sand: 96 h) to ensure that the dry soil particles were well mixed with the added water. The soil water was extracted by mechanical centrifugation and cryogenic vacuum extraction after equilibrium, and their isotope composition was analyzed. Results showed that the isotopic values of soil water extracted by mechanical centrifugation method had no significant difference in same water content with different equilibration times, but were more enriched compared with the reference water isotopic value. The maximum enrichment for hydrogen and oxygen isotope was 7.38‰ and 1.24‰, respectively. In contrast, cryogenic vacuum extraction method resulted in more depleted soil water isotopes than reference water, with the maximum depletion for hydrogen and oxygen isotope being 6.27‰ and 1.03‰, respectively. Moreover, the degree of depletion increased with the increases of equilibrium time (less than 24 h) at low water content, and became stable after 24 h. With the increases of soil water content, the isotopic composition of the extracted soil water was less affected by the two extraction methods. The water isotope value of loam that had high clay content, was more sensitive to the extraction method than the sandy soil that had low clay content. The difference of isotopic composition caused by extraction methods did not affect the plant water source segmentation.


Crowding-induced anisotropic transport modulates reaction kinetics in nanoscale porous media.
We quantify the emergence of persistent anisotropy in the diffusion of spherical tracer particles through a nanoscale porous medium composed of a uniform distribution of purely symmetric crowding particles. We focus on the interior of a biological cell as an example of such a medium and find that diffusion is highly directional for distances comparable to the size of some organelles. We use a geometrical procedure that avoids the standard orientational averaging to quantify the anisotropy of diffusive paths and show that the point source distributions are predominantly of prolate ellipsoidal shape as a result of local volume exclusion. This geometrical symmetry breaking strongly skews the distribution of kinetic rates of diffusion-limited reactions toward small values, leading to the result that, for short to intermediate times, almost 80% of the rates measured in an ensemble of heterogeneous media are smaller than the expected rate in an ideal homogeneous medium of similar excluded volume fraction. This crowding-induced modulation may have implications for our understanding and measurement of diffusion-controlled intracellular reaction kinetics and for experimental nanotechnology applications, such as nanoparticle-based bioimaging and drug delivery, where diffusion plays an important role.


[An epidemic of epileptic seizures after consumption of herbal tea].
At the end of September 2001 the Inspectorate for Health Protection and Veterinary Public Health and the National Poisons Control Centre (NPCC) were informed about adverse health effects after consumption of a herbal tea. During consultations it was suggested that Japanese star anise (Illicium anisatum L.), which is known to contain a neurotoxin, may have been inadvertently mixed into the herbal tea. In view of the severity of the adverse health effects and the clear association with consumption of a specific herbal tea, the supplier was urgently advised to withdraw the suspected herbal tea from the market. A total of 63 persons reported symptoms of general malaise, nausea and vomiting 2-4 hours following consumption of the herbal tea. Twenty-two persons required hospitalisation, of whom 16 due to generalised tonic-clonic seizures. Medical investigations revealed no underlying pathology and after supportive treatment, the patients were discharged in good health. Morphologic and organoleptic investigations of the suspected herbal tea indicated that this possibly contained Japanese star anise. NMR analysis of the herbal tea confirmed the presence of the neurotoxin anisatin, a non-competitive GABA-antagonist which can cause hyperactivity of the central nervous system and tonic-clonic seizures. Ingestion of a herbal tea containing anisatin caused the reported serious adverse health effects. Close cooperation between clinicians, the Inspectorate for Health Protection and Veterinary Public Health and the NPCC played a vital role in preventing further harm to public health.


Dichotic speech tests.
When central auditory dysfunction is present, ability to understand speech in difficult listening situations can be affected. To study this phenomenon, dichotic speech tests were performed with test material in the Swedish language. Digits, spondees, sentences and consonant-vowel syllables were used as stimuli and the reporting was free or directed. The test material was recorded on CD. The study includes a normal group of 30 people in three different age categories; 11 years, 23-27 years and 67-70 years. It also includes two different groups of subjects with suspected central auditory lesions; 11 children with reading and writing difficulties and 4 adults earlier exposed to organic solvents. The results from the normal group do not show any differences in performance due to age. The children with reading and writing difficulties show a significant deviation for one test with digits and one test with syllables. Three of the four adults exposed to solvents show a significant deviation from the normal group.


Social responsibility and research ethics in community-driven studies of industrialized hog production.
Environmental health research can document exposures and health effects that result from inequitable relationships between communities of low income or people of color and the institutions that derive benefits (profits, federal and state funding or services, avoidance of wastes) from activities and policies that burden these communities. Researchers, most of whom work in relatively privileged institutions, are placed in situations of conflicting loyalties if they conduct research in collaboration with, or on behalf of, communities burdened by environmental injustices. These conflicts can threaten the self-interest of researchers and may raise social and ethical issues that do not typically arise in research projects that respond to the agendas of institutions. This article describes how we addressed issues of research ethics and social responsibility in environmental health research on industrialized hog production in North Carolina. Researchers and institutional review boards are not well prepared to address ethical issues when interests of entire communities, as well as individual research participants, are involved. Community-driven research partnerships can help address problems in research ethics and can enhance the social responsibility of researchers and their institutions.


[Expression and significance of Toll like receptor 2 and Toll like receptor 4 in chronic rhinosinusitis].
To explore the role of the innate immune factors TLR2 and TLR4 in the pathogenesis of chronic rhinosinusitis (CRS) by detecting their expression in different clinical types of CRS and the normal control group. Immunohistochemistry was used to detect the expression of TLR2 and TLR4 respectively in 21 cases (chronic rhinosinusitis with nasal polyps, CRSwNP) group, 15 cases (chronic rhinosinusitis without nasal polyos, CRSsNP) group, 11 cases recurrent CRSwNP group and 13 cases control group. Positive cells were counted under the microscope artificially, Mann-Whitney U analysis was applied for the ranked data, and one-way anova analysis was adopted to analyze the experimental group and control group. (1) TLR2 and TLR4 expression had the same characteristics. Expression mainly concentrated in parts of the whole layer of epithelial basement membrane, cytoplasm of glandular cells, very few inflammatory cells such as monocytes and plasma cells in the cytoplasm, sometimes unknown cell nuclei positive expression. (2) The glandular cells were stained manual counting and color grading. TLR2 and TLR4 packet application Wilcoxon rank test Mann-Whitney U test analysis was not statistically significant (P > 0.05), measurement data within the group variance statistical difference between the groups (P < 0.05). The Nasal mucosa can produce the innate immune factors TLR2 and TLR4. The different expression of TLR2 and TLR4 in the various clinical types of CRS suggests that they play the certain role in the pathogenesis of CRS.


SIRT3 functions as a tumor suppressor in hepatocellular carcinoma.
Hepatocellular carcinoma is one of the leading causes for cancer-related mortality worldwide. SIRT3 may function as either oncogene or tumor suppressor in a panel of cancers; however, the role of SIRT3 in hepatocellular carcinoma remains unclear. In this study, we assayed the expression level of SIRT3 in hepatocellular carcinoma tissues by quantitative reverse transcriptase-polymerase chain reaction and immunohistochemistry. A loss-of-function approach was used to examine the effects of SIRT3 on biological activity, including cell proliferative activity and invasive potential. The results demonstrated that the expression levels of SIRT3 protein in hepatocellular carcinoma tissues were significantly downregulated compared with those in adjacent non-cancerous tissues. Furthermore, SIRT3 could decrease cell proliferation and inhibit cell migration/invasion in hepatocellular carcinoma cell line. Taken together, these results elucidated the function of SIRT3 in hepatocellular carcinoma development and suggested that SIRT3 might function as tumor suppressor in hepatocellular carcinoma by targeting PI3K/Akt pathway.


Characterization of an intracellular humanized single-chain antibody to matrix protein (M1) of H5N1 virus.
We developed a human intracellular antibody based on the M1 protein from avian influenza virus H5N1 (A/meerkat/Shanghai/SH-1/2012) and then characterized the properties of this antibody. The M1 protein sequence was amplified by RT-PCR using the cDNA of the H5N1 virus as a template, expressed in bacterial expression system BL21 (DE3) and purified. A human strain, high affinity, and single chain antibody (HuScFv) against M1 protein was obtained by phage antibody library screening using M1 as an antigen. A recombinant TAT-HuScFv protein was expressed by fusion with the TAT protein transduction domain (PTD) gene of HIV to prepare a human intracellular antibody against avian influenza virus. Further analysis demonstrated that TAT-HuScFv could inhibit the hemagglutination activity of the 300 TCID50 H1N1 virus, thus providing preliminary validation of the universality of the antibody. After two rounds of M1 protein decomposition, the TAT-HuScFv antigen binding site was identified as Alanine (A) at position 239. Collectively, our data describe a recombinant antibody with high binding activity against the conserved sequences of avian influenza viruses. This intracellular recombinant antibody blocked the M1 protein that infected intracellular viruses, thus inhibiting the replication and reproduction of H5N1 viruses.


Dopamine recruits D1A receptors to Na-K-ATPase-rich caveolar plasma membranes in rat renal proximal tubules.
Activation of dopamine D(1A) receptors in renal proximal tubules causes inhibition of sodium transporters (Na-K-ATPase and Na/H exchanger), leading to a decrease in sodium reabsorption. In addition to being localized on the plasma membrane, D(1A) receptors are mainly present in intracellular compartments under basal conditions. We observed, using [(3)H]SCH-23390 binding and immunoblotting, that dopamine recruits D(1A) receptors to the plasma membrane in rat renal proximal tubules. Furthermore, radioligand binding and/or immunoblotting experiments using pharmacological modulators showed that dopamine-induced D(1A) receptor recruitment requires activation of cell surface D(1)-like receptors, activation of adenylyl cyclase, and intact endocytic vesicles with internal acidic pH. A key finding of this study was that these recruited D(1A) receptors were functional because they potentiated dopamine-induced [(35)S]GTPgammaS binding, cAMP accumulation, and Na-K-ATPase inhibition. Interestingly, dopamine increased immunoreactivity of D(1A) receptors specifically in caveolin-rich plasma membranes isolated by a sucrose density gradient. In support of this observation, coimmunoprecipitation studies showed that D(1A) receptors interacted with caveolin-2 in an agonist-dependent fashion. The caveolin-rich plasma membranes had a high content of the alpha(1)-subunit of Na-K-ATPase, which is a downstream target of D(1A) receptor signaling in proximal tubules. These results show that dopamine, via the D(1)-like receptor-adenylyl cyclase pathway, recruits D(1A) receptors to the plasma membrane. These newly recruited receptors couple to G proteins, increase cAMP, and participate in dopamine-mediated inhibition of Na-K-ATPase in proximal tubules. Moreover, dopamine-induced recruitment of D(1A) receptors to the caveolin-rich plasma membranes brings them in close proximity to targets such as Na-K-ATPase in proximal tubules of Sprague-Dawley rats.


Insights into mechanisms and proteomic characterisation of Pseudomonas aeruginosa adaptation to a novel antimicrobial substance.
Antibiotic resistance has been reported since the introduction of synthetic antibiotics. Bacteria, such as one of the most common nosocomial pathogens P. aeruginosa, adapt quickly to changing environmental conditions, due to their short generation time. Thus microevolutional changes can be monitored in situ. In this study, the microevolutional process of Pseudomonas aeruginosa PAO1 resistance against a recently developed novel antibacterial zinc Schiff-base (ZSB) was investigated at the proteome level. After extended exposure to ZSB the passaged strain differed in tolerance against ZSB, with the adapted P. aeruginosa PAO1 exhibiting 1.6 times higher minimal inhibitory concentration. Using Two-dimensional Difference Gel Electrophoresis, the changes in the proteome of ZSB adapted P. aeruginosa PAO1 were examined by comparison with the non-adapted P. aeruginosa PAO1. The proteome of the adapted P. aeruginosa PAO1 strain differed significantly from the non-adapted in the abundance of two proteins when both strains were grown under stressing conditions. One protein could be identified as the outer membrane protein D that plays a role in uptake of basic amino acids as well as in carbapeneme resistance. The second protein has been identified as alkyl peroxide reductase subunit F. Our data indicated a slight increase in abundance of alkyl peroxide reductase F (AhpF) in the case of ZSB passaged P. aeruginosa PAO1. Higher abundance of Ahp has been discussed in the literature as a promoter of accelerated detoxification of benzene derivatives. The observed up-regulated AhpF thus appears to be connected to an increased tolerance against ZSB. Changes in the abundance of proteins connected to oxidative stress were also found after short-time exposure of P. aeruginosa PAO1 to the ZSB. Furthermore, adapted P. aeruginosa PAO1 showed increased tolerance against hydrogen peroxide and, in addition, showed accelerated degradation of ZSB, as determined by HPLC measurements.


Iron oxide nanoparticles decrease nuclear fractal dimension of buccal epithelial cells in a time-dependent manner.
In this paper, we present results that iron oxide nanoparticles (IONPs) induce time-dependent structural changes in nuclei of buccal epithelial cells. The cells were treated with magnetite, Fe3 O4 nanoparticles (spherical shape, diameter 80-100 nanometres). The digital micrographs of the nuclei were made in 3 different time points: 15, 30 and 60 min after the treatment with IONPs, as well as in the control cells. A total of 120 nuclear structures (30 per sample) were analysed. Fractal analysis of nuclei was done in ImageJ software of the National Institutes of Health, (Bethesda, MD, USA). For each nuclear structure, the values of fractal dimension and lacunarity were calculated. There was a time-dependent reduction of nuclear fractal dimension in buccal epithelial cells after exposure to magnetite iron oxide nanoparticles. Negative trend was observed (p < 0.01). Nuclear lacunarity, as another fractal parameter was shown to increase, also in a time-dependent manner, after the treatment with IONPs. To our knowledge, this is the first study to investigate effects of magnetite nanomaterial on nuclear fractal complexity, and also the first to apply fractal analysis method in testing of the interaction between nanoparticles and cell nucleus in this experimental setting.


The spatial organization of the human genome.
In vivo, the human genome functions as a complex, folded, three-dimensional chromatin polymer. Understanding how the human genome is spatially organized and folded inside the cell nucleus is therefore central to understanding how genes are regulated in normal development and dysregulated in disease. Established light microscopy-based approaches and more recent molecular chromosome conformation capture methods are now combining to give us unprecedented insight into this fascinating aspect of human genomics.


The Toll-like Receptor 7-Mediated Ro52 Antigen-Presenting Pathway in the Salivary Gland Epithelial Cells of Sjögren's Syndrome.
To investigate whether stimulation with toll-like receptor (TLR) 7 leads to pathways that proceed to tripartite motif-containing protein 21 (TRIM21) or Ro52/SS-A antigen presentation through major histocompatibility complex (MHC) class I in salivary gland epithelial cells (SGECs) from Sjögren's syndrome (SS) patients. Cultured SGECs from SS patients were stimulated with TLR7 agonist, loxoribine, and interferon-β. Cell lysates immunoprecipitated by anti-MHC class I antibody were analyzed by Western blotting. The immunofluorescence of salivary gland tissue from SS and non-SS subjects and cultured TLR7-stimulated SGECs was examined. Significantly increased MHC class I expression was observed in SS patients' ducts versus non-SS ducts; no significant difference was detected for ubiquitin. Upregulated MHC class I in the cell membrane and cytoplasm and augmented Ro52 expression were observed in SGECs stimulated with TLR7. The formation of peptide-loading complex (PLC), including tapasin, calreticulin, transporter associated with antigen processing 1, and endoplasmic reticulum-resident protein 57 in labial salivary glands (LSGs) from SS patients, was dominantly observed and colocalized with MHC class I, which was confirmed in TLR7-stimulated SGEC samples. These findings suggest that the TLR7 stimulation of SS patients' SGECs advances the process toward the antigen presentation of TRIM21/Ro52-SS-A via MHC class I.


Anti-interleukin-31-antibodies ameliorate scratching behaviour in NC/Nga mice: a model of atopic dermatitis.
Interleukin-31 (IL-31), a novel cytokine, is upregulated in atopic dermatitis skin lesions in humans and skin lesions in the NC/Nga mice, a murine model for atopic dermatitis. Here, we investigated the effect of a monoclonal IL-31 antibody on scratching behaviour, weight gain and dermatitis in NC/Nga mice. Mice were divided into three groups, n = 10 in each group. Mice were given monoclonal IL-31 rat-anti-mouse antibody 10 mg/kg or albumin intraperitoneally every fifth day for seven weeks. In addition, the mice in one group were not exposed to any form of intervention. The dermatitis score was clinically assessed twice a week. The scratching behaviour was automatically detected and objectively evaluated. Intervention with IL-31 antibody 10 mg/kg intraperitoneally every fifth day in NC/Nga mice from age 7 weeks reduced the scratching behaviour, but did not have any impact on weight gain or dermatitis. IL-31 antibody reduces scratching behaviour in an atopic dermatitis-like murine model during the onset of clinical skin manifestations. Our findings suggest IL-31 antibody as a new potential therapeutic approach for pruritus in atopic dermatitis and other pruritic diseases.


Polymorphic admixture typing in human ethnic populations.
A panel of 257 RFLP loci was selected on the basis of high heterozygosity in Caucasian DNA surveys and equivalent spacing throughout the human genome. Probes from each locus were used in a Southern blot survey of allele frequency distribution for four human ethnic groups: Caucasian, African American, Asian (Chinese), and American Indian (Cheyenne). Nearly all RFLP loci were polymorphic in each group, albeit with a broad range of differing allele frequencies (delta). The distribution of frequency differences (delta values) was used for three purposes: (1) to provide estimates for genetic distance (differentiation) among these ethnic groups, (2) to revisit with a large data set the proportion of human genetic variation attributable to differentiation within ethnic groups, and (3) to identify loci with high delta values between recently admixed populations of use in mapping by admixture linkage disequilibrium (MALD). Although most markers display significant allele frequency differences between ethnic groups, the overall genetic distances between ethnic groups were small (.066-.098), and < 10% of the measured overall molecular genetic diversity in these human samples can be attributed to "racial" differentiation. The median delta values for pairwise comparisons between groups fell between .15 and .20, permitting identification of highly informative RFLP loci for MALD disease association studies.


Real-time monitoring of the Bragg-peak position in ion therapy by means of single photon detection.
For real-time monitoring of the longitudinal position of the Bragg-peak during an ion therapy treatment, a novel non-invasive technique has been recently proposed that exploits the detection of prompt gamma-rays issued from nuclear fragmentation. Two series of experiments have been performed at the GANIL and GSI facilities with 95 and 305 MeV/u (12)C(6+) ion beams stopped in PMMA and water phantoms. In both experiments, a clear correlation was obtained between the carbon ion range and the prompt photon profile. Additionally, an extensive study has been performed to investigate whether a prompt neutron component may be correlated with the carbon ion range. No such correlation was found. The present paper demonstrates that a collimated set-up can be used to detect single photons by means of time-of-flight measurements, at those high energies typical for ion therapy. Moreover, the applicability of the technique both at cyclotron and at synchrotron facilities is shown. It is concluded that the detected photon count rates provide sufficiently high statistics to allow real-time control of the longitudinal position of the Bragg-peak under clinical conditions.


Novel Endoscopic Transnasal Technique for Anterolateral Thigh Flap Inset Post Nasopharyngeal Carcinoma Resection.
Post-nasopharyngectomy reconstruction for recurrent nasopharyngeal carcinomas has been used for defect coverage and protection of vital structures. With the increasing use of endoscopic transnasal nasopharyngeal extirpation to offset complications faced with open techniques, there is a need for corresponding reconstructive support through a similar approach. We describe a novel endoscopic transnasal anterolateral thigh flap inset technique, combined with a transverse neck incision, in two patients who underwent transnasal nasopharyngectomy. We also include a video presentation of our operative technique. A vastus lateralis fascia free flap was used for one patient, and a vastus lateralis muscle free flap for the other. Both patients were aged 51 years. Mean nasopharyngeal defect size was 20 cm2 (range 12-28 cm2). Average surgical stay was 13.5 days (11-16 days) and flap mucosalization was complete for both patients. No recipient site complications were observed in either patient, although donor site seroma formation was seen in one patient. Average time to speech recovery was 1.5 months (range 1-2 months) for both patients. Time to diet recovery was 2 months for one patient, whereas the other was on long-term percutaneous endoscopic gastrostomy feeding. There were no flap failures or peri-operative mortalities. Endoscopic transnasal anterolateral thigh flap inset to reconstruct the nasopharyngeal space is an effective technique that confers reduced morbidity and potentially better outcomes compared with open techniques.


Brasilianoids A-F, New Meroterpenoids From the Sponge-Associated Fungus Penicillium brasilianum.
3,5-Dimethylorsellinic acid (DMOA) derived meroterpenoids comprise an unique class of natural products with diverse scaffolds and with a broad spectrum of bioactivities. Bioinformatics analysis of the gene clusters in association with the qRT-PCR detection of the amplification of two key genes led to speculate that the sponge associated fungus Penicillium brasilianum WZXY-m122-9 is a potential producer of meroterpenoids. Chromatographic separation of the EtOAc extract of this fungal strain on a large-scale fermentation resulted in the isolation of six new DMOA-related meroterpenoids with trivial names of brasilianoids A-F (1-6), together with preaustinoid D and preaustinoid A2. The structures were determined by extensive analyses of spectroscopic data, including the X-ray diffraction and the ECD data for configurational assignment. Brasilianoids A and F showed an unprecedented skeleton with a γ-lactone in ring A, while brasilianoids B-C featured a 7/6/6/5/5 pentacyclic ring system finding in nature for the first time. The biosynthetic relationship among the isolated compounds was postulated. Compound 1 significantly stimulated the expression of filaggrin and caspase-14 in HaCaT cells in dose-dependent manner, while compounds 2 and 3 showed moderate inhibition against NO production in LPS-induced RAW 264.7 macrophages.


[Marking bourdaries as a strategy of social control: the case of homicidal violence in Medellin, Colombia].
As part of a research study undertaken in the period 2003-2011 to understand situations of homicidal violence based in perceptions regarding the act of violence and the surrounding context, we reflect on the meaning of "invisible bourdaries" in the neighborhoods of Medellin (Colombia). Using a qualitative approach that combines documentary sources and interviews, the experiences of 8 participants are analyzed. In the primary results we can see how control over neighborhoods is exercised by different actors through bourdaries not visible to ordinary people. Nevertheless, around these lines people are recruited and controlled and strategies to illegally generate economic resources and to regulate the cultural and social activities of inhabitants are consolidated, thus affecting the social dynamics and imaginary of the neighborhood. In this way, the territories, friendships, and affects of young victims – who are not linked to illegal groups and/or do not have "information" – and of defenseless older adults are controlled.


Fluorescein Isothiocyanate Labeling Antigen-Based Immunoassay Strip for Rapid Detection of Acidovorax citrulli.
A simple and fast immunoassay strip to detect Acidovorax citrulli (Ac) using fluorescein isothiocyanate as a marker was developed. Fluorescein isothiocyanate (FITC) was added to sample culture medium for bacteria incubation, and the bacteria could emit a yellow-green fluorescence under ultraviolet light and become a fluorescent probe. This immunofluorescence strip (IFS) was based on the binding between fluorescent bacteria and the unlabeled monoclonal antibody (McAb) immobilized on the test area in nitrocellulose membrane. The detection limit of the strip was 106 CFU/ml with a result that could be observed within 10 min. The IFS could detect eight strains of Ac and display no cross-reactions with 30 other pathogenic strains. The detection results would not be affected by impurities in plant or unknown microorganisms in natural field samples and were consistent with PCR results, indicating that the IFS has high accuracy. This is the first report of using only one unlabeled McAb to develop a direct-type immunofluorescence strip for the rapid detection of Ac. The IFS reduced detection time and simplified operation procedures compared with the traditional enzyme-linked immunosorbent assay (ELISA) and PCR methods. In addition, this simple and inexpensive method will play a significant role in monitoring plant pathogens on field detection.


Study and Therapeutic Progress on Intracranial Serpentine Aneurysms.
An intracranial serpentine aneurysm (SA) is a clinically rare entity, and very few multi-case studies on SA have been published. The present study reviewed the relevant literature available on PubMed. The studied information included the formation mechanism and natural history of SA as well as its clinical manifestation, imaging characteristics, and current treatments. After reviewing the literature, we conclude that intracranial SA can be managed surgically and by endovascular embolization, but the degree of blood flow in normal brain tissue distal to the SA must be evaluated. A balloon occlusion test (BOT) or cross compression test is recommended for this evaluation. If the collateral circulation is sufficiently compensatory, direct excision or embolization can be performed. However, if the compensatory collateral circulation is poor, a bypass surgery is necessary. Satisfactory results can be achieved in the majority of SA patients after treatment. However, the size of the aneurysm may increase in some patients after endovascular treatment. Special attention should be paid to cases exhibiting a significant mass effect to avoid subsequent SA excision due to an intolerable mass effect. Satisfactory results can be achieved with careful treatment of SA.


[Mobility and Fall Prevention in older Community-dwelling Persons].
An increasing proportion of oldest old persons (over 80 years) accompanies the demographic change. Furthermore, the gender ration will also change and the percentage of older men will increase. Mobility is an important factor for maintaining independence and for "healthy aging". Mobility has to be regarded in the context of individual and environmental factors. With increasing age, the risk of mobility limitation increases. Next to mobility, falls are also a drastic experience for older persons and cannot only have physical consequences like injuries but also psychological consequences as fall-related psychological concerns. These psychological consequences can produce negative behavior e. g. reducing physical activity.


[Surgical treatment for metastatic gastric cancer].
The incidence of metastatic gastric cancer (MGC) is high and the prognosis is poor with 5-year survival rarely exceeding 5%. Systemic chemotherapy is the cornerstone for MGC in current treatment guidelines. Recent studies have revealed both reduction surgery and conversion surgery can improve patient's prognosis, indicating the value of surgical treatment in MGC. Based on the present therapeutic strategy, we discuss the indication, extent and timing of surgery for MGC. Our conclusions are as followings: the treatment of gastric cancer is chemotherapy-based, target-included and surgery-combined multidisciplinary therapy; patients with single non-curable factor are most likely to benefit from surgery-combined multidisciplinary therapy; the optimal timing for surgery depends on the response to chemotherapy; it is worthwhile to explore the conversion therapy combining target therapy and chemotherapy; the resection of metastasis lesion can be managed according to Japanese guideline; in order to establish high-level evidence, it is necessary to unite experts from different disciplines to conduct clinical trial according to the category of metastasis of gastric cancer.


ATG4B contains a C-terminal LIR motif important for binding and efficient cleavage of mammalian orthologs of yeast Atg8.
The cysteine protease ATG4B cleaves off one or more C-terminal residues of the inactive proform of proteins of the ortholog and paralog LC3 and GABARAP subfamilies of yeast Atg8 to expose a C-terminal glycine that is conjugated to phosphatidylethanolamine during autophagosome formation. We show that ATG4B contains a C-terminal LC3-interacting region (LIR) motif important for efficient binding to and cleavage of LC3 and GABARAP proteins. We solved the crystal structures of the GABARAPL1-ATG4B C-terminal LIR complex. Analyses of the structures and in vitro binding assays, using specific point mutants, clearly showed that the ATG4B LIR binds via electrostatic-, aromatic HP1 and hydrophobic HP2 pocket interactions. Both these interactions and the catalytic site-substrate interaction contribute to binding between LC3s or GABARAPs and ATG4B. We also reveal an unexpected role for ATG4B in stabilizing the unlipidated forms of GABARAP and GABARAPL1. In mouse embryonic fibroblast (MEF) atg4b knockout cells, GABARAP and GABARAPL1 were unstable and degraded by the proteasome. Strikingly, the LIR motif of ATG4B was required for stabilization of the unlipidated forms of GABARAP and GABARAPL1 in cells.


Comparison of the 8th union for international cancer control lymph node staging system for gastric cancer with two other lymph node staging systems.
The log odds of positive lymph nodes (LODDS) and the metastatic lymph node ratio (MLR) staging systems have previously been demonstrated to exhibit advantages compared with the tumor-node-metastasis (TNM) staging system in predicting the prognosis of gastric cancer. The current study compared the prognostic significance of the newest Union for International Cancer Control Node classification with the LODDS and MLR staging systems. From September 2010 to December 2012, all medical records for patients with gastric cancer at the Third Affiliated Hospital of Soochow University were retrospectively analyzed and the clinicopathologic characteristics were reviewed. Cut-off points were selected to divide the patients with gastric cancer into different groups. Univariate and multivariate analyses were performed to identify the prognostic risk factors for gastric cancer. The Harrell's concordance index (C-index) was adopted to compare the prognostic value of the three staging systems. A total of 877 patients with gastric cancer who met the inclusion criteria were analyzed in the current study. The patients were classified according to the three MLR subgroups as follows: MLR0 (MLR=0), MLR1 (0<MLR≤0.28) and MLR2 (0.28<MLR<1). The patients were classified according to the LODDS subgroups as follows: LODDS1 (LODDS≤-0.5), LODDS2 (-0.5<LODDS≤0), LODDS3 (0<LODDS≤0.5) and LODDS4 (LODDS>0.5). Based on multivariate analysis, LODDS, MLR and pathological node (pN) stage could significantly predict survival rates of patients with gastric cancer. According to the C-index, the LODDS staging system more accurately predicted the 5-year overall survival for patients with gastric cancer compared with the other two staging systems. In summary, the current study has identified that LODDS may be superior to the MLR and pN staging systems in predicting the prognosis of patients with gastric cancer. However MLR may exhibit advantages compared with LODDS for patients who have undergone adequate lymphadenectomies.


[Evaluation of the therapeutic benefits in relation to the extent of surgery in patients with differentiated thyroid carcinoma].
Evaluation of the therapeutic benefits in relation to the stage of thyroid cancer and to the extent of surgery and the risk of postoperative complications. Retrospective analysis of differentiated thyroid carcinoma (DTC) patients staged T1M0 versus T2-T4M0 was performed. All of them were treated or diagnosed in Institute of Oncology in Gliwice between 1986-1998. Previously they were operated in various surgical centers all-over Poland. The risk of death, local relapse and postoperative complications were analyzed using the decisiontree model to evaluate the therapeutic benefits. The recurrent laryngeal nerve injury (transient or permanent) was observed in retrospective analysis in 21% of patients, while postoperative hyperparathyroidism in 15.8%. The analysis of the therapeutic benefit index showed no advantage of total thyroidectomy in the T1M0 group (0.96 vs. 0.98 in patients treated by less than total thyroidectomy). The advantage of radical surgery was confirmed in T2-T4M0 group. The therapeutic benefit index was 0.92 in patients treated by total thyroidectomy and 0.69 in those who received less extensive operation. The analysis of therapeutic benefits confirmed the limit of 1 cm tumor diameter between less extensive surgery and total thyroidectomy. It showed that total thyroidectomy brings a significant therapeutic benefits in patients in > T1M0 stage. The improvement of overall survival and decrease of local relapse far outweigh the disadvantages related to postoperative complications.


Perspectives and Experiences of Parents and Adolescents Who Participate in a Pediatric Precision Oncology Program: "When You Feel Helpless, This Kind of Thing Is Very Helpful".
The SickKids Cancer Sequencing (KiCS) Program, launched in 2016, evaluates the clinical utility of paired tumor/germline Next-Generation Sequencing (NGS) in pediatric oncology patients with hard-to-cure and rare cancers. In anticipation of further widespread adoption of NGS, we aimed to characterize the experiences and perspectives of adolescents and parents of patients who have already undergone NGS evaluation, focusing on the psychosocial impact and personal utility. Parents of patients with pediatric cancer and adolescent patients who have participated in KiCS were invited to participate in semistructured interviews. Transcripts were analyzed using an inductive content analytic approach. Of 45 individuals invited, 22 parents and 10 adolescents were interviewed (71% response rate). Prominent psychosocial themes were low distress, relief, and sense of control; some expressed fear of the unknown. In exploring constructs of personal utility, parents highlighted hope for treatment options despite low expectations for results with clinical impact, whereas adolescents articulated altruistic motivations and less hope for personal clinical benefit. Bringing closure and answering the question of why the cancer occurred was a salient theme among both groups. Both parents and adolescents find benefit and clear decisional satisfaction with participation. No participants expressed regret. This study suggests that parents and adolescents benefit from NGS evaluation beyond the return of clinically relevant results. Our findings lay the framework for future work evaluating the value of NGS in pediatric precision oncology care through assessment of patient-reported outcomes and experiences. These results also guide provision of pre- and post-test education and support, which will facilitate patient-centered delivery of NGS practices.


Mutational Profile from Targeted NGS Predicts Survival in LDCT Screening-Detected Lung Cancers.
The issue of overdiagnosis in low-dose computed tomography (LDCT) screening trials could be addressed by the development of complementary biomarkers able to improve detection of aggressive disease. The mutation profile of LDCT screening-detected lung tumors is currently unknown. Targeted next-generation sequencing was performed on 94 LDCT screening-detected lung tumors. Associations with clinicopathologic features, survival, and the risk profile of a plasma microRNA signature classifier were analyzed. The mutational spectrum and frequency observed in screening series was similar to that reported in public data sets, although a larger number of tumors without mutations in driver genes was detected. The 5-year overall survival (OS) rates of patients with and without mutations in the tumors were 66% and 100%, respectively (p = 0.015). By combining the mutational status with the microRNA signature classifier risk profile, patients were stratified into three groups with 5-year OS rates ranging from 42% to 97% (p < 0.0001) and the prognostic value was significant after controlling for stage (p = 0.02). Tumor mutational status along with a microRNA-based liquid biopsy can provide additional information in planning clinical follow-up in lung cancer LDCT screening programs.


Wait times for breast cancer care.
Measurement of care time intervals is complex, being influenced by many factors. The definition of the care interval monitored can also bias the detection of changes in waits. The implications of using different care interval definitions to report wait times and identify delays in care provision were examined using a retrospective chart review of 637 women with surgically treated breast cancer who were referred to a cancer centre between September 1999 and 2000 or September 2003 and 2004. Overall waits between detection and adjuvant treatment increased by 12 days over the two periods, but their exact location and cause(s) could not be determined at such a low-resolution interval. At higher resolutions of care intervals, reporting the comprehensive sequence of care events, the prolongation was mainly associated with delayed access to surgery (4 days) and delivery of adjuvant chemotherapy (4 days). The latter went unnoticed when waits were reported at intermediate (referral to adjuvant treatment) and low (detection to adjuvant treatment) resolutions. Disease stage and type of first adjuvant treatment consistently and significantly influenced the length of waits. Comprehensive monitoring of the entire care path is essential to effectively prioritize interventions, assess their outcomes and optimise access to cancer care.


Aging and endothelial progenitor cell telomere length in healthy men.
Telomere length declines with age in mature endothelial cells and is thought to contribute to endothelial dysfunction and atherogenesis. Bone marrow-derived circulating endothelial progenitor cells (EPCs) are critical to vascular health as they contribute to both reendothelialization and neovascularization. We tested the hypothesis that EPC telomere length decreases with age in healthy adult humans. Peripheral blood samples were collected from 40 healthy, non-obese, sedentary men: 12 young (age 21-34 years), 12 middle-aged (43-55 years) and 16 older (57-68 years). Putative EPCs were isolated from peripheral blood mononuclear cells and telomere length was determined using genomic DNA preparation and Southern hybridization techniques. EPC telomere length (base pairs) was approximately 20% (p=0.01) lower in the older (8492+523 bp) compared to the middle-aged (10,565+572 bp) and young (10,205+501 bp) men. Of note, there was no difference in EPC telomere length between the middle-aged and young men. These results demonstrate that EPC telomere length declines with age in healthy, sedentary men. Interestingly, telomere length was well preserved in the middle-aged compared to young men, suggesting that EPC telomere shortening occurs after the age of 55 years.


A Prognostic Factor for Prolonged Mechanical Ventilator-Dependent Respiratory Failure after Cervical Spinal Cord Injury : Maximal Canal Compromise on Magnetic Resonance Imaging.
The period of mechanical ventilator (MV)-dependent respiratory failure after cervical spinal cord injury (CSCI) varies from patient to patient. This study aimed to identify predictors of MV at hospital discharge (MVDC) due to prolonged respiratory failure among patients with MV after CSCI. Two hundred forty-three patients with CSCI were admitted to our institution between May 2006 and April 2018. Their medical records and radiographic data were retrospectively reviewed. Level and completeness of injury were defined according to the American Spinal Injury Association (ASIA) standards. Respiratory failure was defined as the requirement for definitive airway and assistance of MV. We also evaluated magnetic resonance imaging characteristics of the cervical spine. These characteristics included : maximum canal compromise (MCC); intramedullary hematoma or cord transection; and integrity of the disco-ligamentous complex for assessment of the Subaxial Cervical Spine Injury Classification (SLIC) scoring. The inclusion criteria were patients with CSCI who underwent decompression surgery within 48 hours after trauma with respiratory failure during hospital stay. Patients with Glasgow coma scale 12 or lower, major fatal trauma of vital organs, or stroke caused by vertebral artery injury were excluded from the study. Out of 243 patients with CSCI, 30 required MV during their hospital stay, and 27 met the inclusion criteria. Among them, 48.1% (13/27) of patients had MVDC with greater than 30 days MV or death caused by aspiration pneumonia. In total, 51.9% (14/27) of patients could be weaned from MV during 30 days or less of hospital stay (MV days : MVDC 38.23±20.79 vs. MV weaning, 13.57±8.40; p<0.001). Vital signs at hospital arrival, smoking, the American Society of Anesthesiologists classification, Associated injury with Injury Severity Score, SLIC score, and length of cord edema did not differ between the MVDC and MV weaning groups. The ASIA impairment scale, level of injury within C3 to C6, and MCC significantly affected MVDC. The MCC significantly correlated with MVDC, and the optimal cutoff value was 51.40%, with 76.9% sensitivity and 78.6% specificity. In multivariate logistic regression analysis, MCC >51.4% was a significant risk factor for MVDC (odds ratio, 7.574; p=0.039). As a method of predicting which patients would be able to undergo weaning from MV early, the MCC is a valid factor. If the MCC exceeds 51.4%, prognosis of respiratory function becomes poor and the probability of MVDC is increased.


5-Aminolevulinic acid strongly enhances delayed intracellular production of reactive oxygen species (ROS) generated by ionizing irradiation: quantitative analyses and visualization of intracellular ROS production in glioma cells in vitro.
Postoperative adjuvant radiotherapy has important roles in multimodal treatment for highly aggressive malignant gliomas. Previously, we demonstrated that multi-dose ionizing irradiation with repetitive administration of 5-aminolevulinic acid (5-ALA) enhanced the host antitumor response and strongly inhibited tumor growth in experimental glioma. However, the mechanism of the radiosensitizing effect of 5-ALA is not known. Ionizing irradiation not only causes reactive oxygen species (ROS) formation initially by water radiolysis but also induces delayed production of mitochondrial ROS for mediating the long-lasting effects of ionizing irradiation on tumor cells. 5-ALA leads to high accumulation of protoporphyrin IX (PpIX) in the mitochondria of tumor cells, yet can also improve dysfunction of the mitochondrial respiratory chain in tumor cells. Here, we assessed the effect of 5-ALA-induced PpIX synthesis and delayed production of intracellular ROS after ionizing irradiation with 5-ALA in glioma cells in vitro. Temporal changes in intracellular 5-ALA-induced PpIX synthesis after ionizing irradiation in glioma cell lines were evaluated using flow cytometry (FCM). Then, the effect of 5-ALA on delayed production of intracellular ROS 12 h after ionizing irradiation in glioma cells was evaluated by FCM and confocal laser scanning microscopy. Ionizing irradiation had no effect on 5-ALA-induced PpIX synthesis in glioma cells. Delayed intracellular production of ROS was significantly higher than that just after ionizing irradiation, but 5-ALA pretreatment strongly enhanced the delayed intracellular production of ROS, mainly in the cytoplasm of glioma cells. This 5-ALA-induced increase in the delayed production of ROS tended to be higher in the case of 5-ALA treatment before rather than after ionizing irradiation. These results suggest that 5-ALA can affect tumor cells under ionizing irradiation, and greatly increase secondary intracellular production of ROS long after ionizing irradiation, thereby causing a radiosensitizing effect in glioma cells.


The enhancement of the immune response by pain stimulation in mice. I. The enhancement effect on PFC production via sympathetic nervous system in vivo and in vitro.
Effects of catecholamines and osmotical and physical stimuli on the induction of anti-sheep red blood cells (SRBC) plaque-forming cells (PFC) were investigated in (C57BL/6 X BALB/c)F1 mice in vivo and in vitro. The anti-SRBC PFC from mice immunized with 5 X 10(7) SRBC was markedly increased by daily s.c. injections of epinephrine. The enhancement of PFC by epinephrine was completely blocked by preadministration with propranolol and hexamethonium, but not with phentolamine. The PFC was increased by osmotic and physical stimuli given once a day for 4 days after immunization with SRBC. The enhancement of PFC by these stimuli was completely blocked by preadministration with propranolol and hexamethonium. The enhancement of PFC by physical stimuli was observed in nonimmunized mice when spleen cells from stimulated mice were cultured with SRBC in vitro. In normal mice, the enhancement of PFC was observed 2 hr after one physical stimulation. However, spleen cells from mice given two physical stimuli did not show the enhancement of PFC after treatment with anti-Thy-1.2 antibody and complement, nor after removal of nonadherent cells. Next, the serum obtained from mice 30 to 60 min after a physical stimulation enhanced PFC of normal mice spleen cells in vitro, but the enhancement was abolished by the addition of propranolol. The enhancement of anti-SRBC PFC by s.c. injection of epinephrine suggested that the autonomic nervous system, especially the sympathetic nervous system, was activated by a local stimulus effect of the injection. This enhancement of anti-SRBC PFC appear to be due to the activation of antigen non-specific helper T lymphocytes by the beta-actin of endogenous catecholamines from the adrenal gland.


Genetics of sarcoidosis: candidate genes and genome scans.
Human leukocyte antigen class II allele associations and T-cell receptor beta chain bias in sarcoidosis suggest a specific disease-triggering antigen exposure in a genetically susceptible host. The cause of sarcoidosis has been elusive, but genetics provides one of the few promising avenues to further our understanding. We review the association studies and genome scans used to identify the genes involved in sarcoidosis.


Zinc supplementation improved cognitive performance and taste acuity in Indian adolescent girls.
To test the efficacy of zinc supplementation through diet or ayurvedic zinc tablet on cognitive function and taste acuity in adolescent girls. Using zinc-rich food items, snacks were prepared by adopting food-processing methods that enhance zinc bioavailability. Ayurvedic zinc tablet (jasad bhasma) was chosen as a natural elemental zinc supplement. Efficacy of snacks and the tablet was assessed in 180 schoolgirls (12.5 +/- 0.85 years) from Pune City, India, who were randomly allocated to any of the 3 groups: (1) ayurvedic zinc tablet-J, (2) zinc-rich snacks-D, or (3) Control-C. Supplementation was given on every school day (6 days/wk) for 10 weeks. All measurements were recorded at baseline and at the end of the study period. Food intake was recorded by 24-hour diet recall on 3 random days. Hemoglobin, serum ferritin, and plasma zinc were estimated on a fasting blood sample. Cognitive assessment was done on each participant using tests for simple reaction time (SRT), recognition reaction time (RRT), visual memory, and Raven's Progressive Matrices (RPM). Taste acuity was determined by recognition thresholds for salt (RTS) over a range of 10 different salt concentrations. A higher increase in plasma zinc (61.3%) was observed in the J group than in the D group (9.9%) (p < 0.01), whereas plasma zinc declined in the control group (-2.2%) over baseline (p > 0.1). Hemoglobin showed no change in all 3 groups (p > 0.1). Percent increment in scores for memory and RPM was significantly more in the D and J groups (24.5%-29.6%) than in the C group (6.5%) (p < 0.05). Mean SRT and RRT were reduced more in the D and J groups (5%-16%) than in the C group (1.6%) (p < 0.05). A significant fall in median RTS from 5 to 2.5 mmol/L was noted after both diet and zinc supplementation (p < 0.01); however, it remained the same at 5 mmol/L in the Control group after 10 weeks. Supplementation of ayurvedic zinc and zinc-rich foods are effective in improving cognitive performance and the recognition threshold for salt of adolescent girls.


Preparing Silver Linings for a Cloudy Day: The Consequences of Preemptive Benefit Finding.
Waiting for important news is stressful. In four studies, we assess the utility of preemptive benefit finding, a coping strategy in which people seek silver linings in bad news before receiving news, for emotional well-being across several waiting periods (waiting for bar exam results, the outcome of political elections, and results of a fictitious health risk assessment). Our findings support the effectiveness of preemptive benefit finding while waiting, such that identifying benefits in bad news while waiting predicts more positive emotions during the wait (Studies 3 and 4) and buffers people against the emotional consequences of bad news by boosting post-news positive emotions (Studies 2-4). Importantly, engaging in preemptive benefit finding does not backfire if a person ultimately receives good news (Studies 1, 3, and 4). We discuss results from a mini meta-analysis and consider implications of our findings for interventions to improve well-being while waiting and after news arrives.


Resistance Risk Assessment of Spodoptera frugiperda (Lepidoptera: Noctuidae) and Diatraea saccharalis (Lepidoptera: Crambidae) to Vip3Aa20 Insecticidal Protein Expressed in Corn.
Transgenic Agrisure Viptera 3 corn that expresses Cry1Ab, Vip3Aa20, and EPSPS proteins and Agrisure Viptera expressing Vip3Aa20 are used for control of Spodoptera frugiperda (J.E. Smith) and Diatraea saccharalis (F.) in Brazil. To support a resistance management program, resistance risk assessment studies were conducted to characterize the dose expression of Vip3Aa20 protein and level of control against these species. The Vip3Aa20 expression in Agrisure Viptera 3 and Agrisure Viptera decreased from V6 to V10 stage of growth. However, Vip3Aa20 expression in Agrisure Viptera 3 at V6 and V10 stages was 13- and 16-fold greater than Cry1Ab, respectively. The Vip3Aa20 expression in lyophilized tissue of Agrisure Viptera 3 and Agrisure Viptera diluted 25-fold in an artificial diet caused complete larval mortality of S. frugiperda and D. saccharalis. In contrast, lyophilized tissue of Bt11 at the same dilution does not provide complete mortality of these species. Agrisure Viptera 3 and Agrisure Viptera also caused a high level of mortality against S. frugiperda and D. saccharalis. Moreover, 100% mortality was observed for S. frugiperda larvae (neonates through fifth-instar larvae) when fed in corn with the Vip trait technology. Viptera corn achieves a high level of control against S. frugiperda and D. saccharalis providing a high dose, which is an important determination to support the refuge strategy for an effective resistance management program.


Dental health changes in an Australian Defence Force population.
The dental histories of 100 long-term members of the Royal Australian Air Force were obtained from their dental records. Data relating to dental health status, as measured by the DMF indices and the T-Health scores, were tested to determine if there were any correlations present between changes in dental health and certain risk factors which had been identified by previous studies in the General Dental Service in Scotland. These studies had demonstrated positive relationships between the frequency of dental examination and the frequency with which patients changed their dentist, with the amount of dental treatment the patients received. However, within the potential limitations imposed by the sample size, the present long-term study did not detect any significant associations between the frequencies of dental examination or change of dentist, with changes in dental health.


Outcomes in Patients with Liver Dysfunction Post SARS-CoV-2 Infection: What Should We Measure?
Since 2019, the COVID-19 pandemic wreaked havoc all over the world. Early in the course of the pandemic, multiple hepatic manifestations of COVID-19 were noted. We aim to categorize hepatic dysfunction and its outcome in COVID-19 infection. This is a review article based on a literature search in PubMed and Medline databases for articles detailing short-term and long-term outcomes of COVID-19 related liver dysfunction. The most common hepatic manifestation of COVID-19 was aspartate amino transferase (AST) predominant transaminase elevation. Transaminases improve once the COVID-19 infection resolves. In addition, COVID-19 cholangiopathy, autoimmune hepatitis associated COVID-19, and splanchnic venous thrombosis triggered by COVID-19 are other manifestations. Patients with preexisting liver disease, especially those with cirrhosis, have poor prognosis with COVID-19 infections compared to the general population. Elevations in liver tests were associated with severe COVID-19 infections. Patients with chronic liver disease have a higher risk of morbidity and mortality from COVID-19 infection. Among patients with chronic liver disease, decompensated liver cirrhosis, hepatocellular carcinoma and alcohol-associated liver disease were associated with an increased risk of severity and mortality from COVID-19 infection. Interactions between antiviral therapy for COVID-19 and hepatitis B/hepatitis C medications must be considered in patients with chronic viral hepatitis and COVID-19 infection. COVID-19 vaccination-related hepatic dysfunction has been reported. COVID-19 is here to stay. Hepatic dysfunction in COVID-19 signals severe COVID-19 infections. Patients with chronic liver disease have higher mortality from COVID-19 than general population. It is important to remember the lessons learned throughout the covid pandemic to take care of patients with COVID-19 now and in the future. Further studies are needed to document long-term outcomes in patients with COVID-19 who developed hepatic dysfunction.


Acylation - A New Means to Control Traffic Through the Golgi.
The Golgi is well known to act as center for modification and sorting of proteins for secretion and delivery to other organelles. A key sorting step occurs at the trans-Golgi network and is mediated by protein adapters. However, recent data indicate that sorting also occurs much earlier, at the cis-Golgi, and uses lipid acylation as a novel means to regulate anterograde flux. Here, we examine an emerging role of S-palmitoylation/acylation as a mechanism to regulate anterograde routing. We discuss the critical Golgi-localized DHHC S-palmitoyltransferase enzymes that orchestrate this lipid modification, as well as their diverse protein clients (e.g., MAP6, SNAP25, CSP, LAT, β-adrenergic receptors, GABA receptors, and GLUT4 glucose transporters). Critically, for integral membrane proteins, S-acylation can act as new a "self-sorting" signal to concentrate these cargoes in rims of Golgi cisternae, and to promote their rapid traffic through the Golgi or, potentially, to bypass the Golgi. We discuss this mechanism and examine its potential relevance to human physiology and disease, including diabetes and neurodegenerative diseases.


Local site preference rationalizes disentangling by DNA topoisomerases.
To rationalize the disentangling action of type II topoisomerases, an improved wormlike DNA model was used to delineate the degree of unknotting and decatenating achievable by selective segment passage at specific juxtaposition geometries and to determine how these activities were affected by DNA circle size and solution ionic strength. We found that segment passage at hooked geometries can reduce knot populations as dramatically as seen in experiments. Selective segment passage also provided theoretical underpinning for an intriguing empirical scaling relation between unknotting and decatenating potentials.


Measuring public attitudes on abortion: methodological and substantive considerations.
Data from a 1989 CBS News/New York Times survey are used to examine the effect that the framing of questions on abortion has on estimates of what proportions of the population support various legal positions. The nationwide data and results from six state polls show that general questions with only two or three options overestimate the proportions of respondents who either favor a ban on all abortion or who would allow abortion under all circumstances. Questions that pose specific circumstances result in movement of respondents out of extreme categories and into more moderate ones. Even respondents who indicate they would favor abortion in all specific circumstances and those who favor abortion in none are likely to moderate their views when asked if they support restrictions that have been proposed in a number of states.