Encapsulation of tea polyphenols into high amylose corn starch composite nanofibrous film for active antimicrobial packaging.
Starch-based composite nanofibrous films loaded with tea polyphenols (TP) were successfully fabricated through electrospinning high amylose corn starch (HACS) with aid of polyvinyl alcohol (PVA), referred as HACS/PVA@TP. With the addition of 15 % TP, HACS/PVA@TP nanofibrous films exhibited enhanced mechanical properties and water vapor barrier capability, and their hydrogen bonding interactions were further evidenced. TP was slowly released from the nanofibrous film and followed Fickian diffusion mechanism, which achieved the controlled sustained release of TP. Interesting, HACS/PVA@TP nanofibrous films effectively improved antimicrobial activities against Staphylococcus aureus (S. aureus) and prolonged the shelf life of strawberry. HACS/PVA@TP nanofibrous films showed superior antibacterial function by by destroying cell wall and cytomembrane, and degrading existing DNA fragments, stimulating excessive intracellular reactive oxygen species (ROS) generation. Our study demonstrated that the functional electrospun Starch-based nanofibrous films with enhanced mechanical properties and superior antimicrobial activities were potential for the application in active food packaging and relative areas.


Effect of cup medialization on primary stability of press-fit acetabular cups.
Appropriate restoration of the native centre of rotation is of paramount importance in total hip arthroplasty. Reconstruction of the centre of rotation depends on reaming technique: conventional approaches require more cup medialization than anatomical preparations. To date, the influence of cup medialization on socket stability in cementless implants is still unknown. Ten cadaveric hemipelvises were sequentially reamed using anatomical technique (only subchondral bone removal with restoration of the native centre of rotation) and conventional preparation (reaming to the lamina and medializing the cup). A biomechanical test was performed on the reconstructions. Implant motions were measured with digital image correlation while a cyclic load of increasing magnitude was applied. No significant difference was measured between the two implantation techniques in terms of permanent cup migrations. The only significant difference was found for the cup inducible rotations, where the conventional technique was associated with larger rotations. Conventional reaming and cup medialization do not improve initial cup stability. Beyond the recently questioned concerns about medialization and hip biomechanics, this is another issue to bear in mind when reaming the acetabulum.


An international comparison of Google searches for sunscreen, sunburn, skin cancer, and melanoma: Current trends and public health implications.
Google Trends® (GT) offers insight into public interests and behaviors and holds potential for guiding public health campaigns. This study explored international trends in English-speaking countries (United States, United Kingdom, Canada, Australia, New Zealand) in searches for sunscreen, sunburn, skin cancer, and melanoma to better guide skin cancer prevention campaigns. Google Trends® was queried for search terms from January 1, 2004 to December 31, 2017. Respective national databases were queried for melanoma outcome data from 2004 to 2014 and compared with time-matched search data. Correlations between search terms, time, and melanoma outcomes were assessed for each country. Quantitative analyses were performed to evaluate differences in search volumes between countries with varying melanoma incidence. In all countries, the strongest intra-term correlation was between sunscreen and sunburn. Searches for sunscreen and sunburn are increasing for all countries. For all countries except the United Kingdom and New Zealand, searches for skin cancer and melanoma are decreasing for one or both terms. Correlations between search terms and melanoma outcomes were variable and specific to each country. Quantitative analysis revealed that countries with higher melanoma incidence had higher search volumes for all terms. Search volumes were especially high for skin cancer and melanoma in Australia compared with other countries. Comparisons between moderate melanoma incidence countries were less clear. Online skin cancer prevention campaigns should focus on the search terms sunburn and sunscreen, especially given the declining interest between 2004 and 2016 in the terms skin cancer and melanoma seen in multiple countries. Search term interests varied with melanoma outcomes and between countries, suggesting the importance of customizing approaches based on local population interests and geographic areas.


Development of multicolor carbon nanoparticles for cell imaging.
Multicolor carbon nanoparticles (CNPs) were prepared, characterized and developed as fluorescent probes for cell imaging. The fluorescent CNPs were prepared with a facile hydrothermal oxidation route by using linear polysaccharide cellulose and cyclic oligosaccharide cyclodextrin as carbon sources. The characterizations by transmission electron microscopy show that the prepared cellulose-CNPs and cyclodextrin-CNPs are spherical, well-dispersed in water with average diameters of 100 nm and 76 nm, respectively. Under the excitation of UV light, the CNPs are strongly luminescent with an excitation-dependent emission behavior and bathochromic emission properties. The fluorometric methods show that the cellulose-CNPs and cyclodextrin-CNPs are strongly fluorescent with fluorescence quantum yield of 7.47% and 4.49%, respectively. The multicolor CNPs have excellent photostability toward photobleaching. Strong near-infrared fluorescence of the carbon nanoparticles was observed with a 632.8 nm excitation wavelength laser. The oxidative metal ions like Hg(II), Cu(II) and Fe(III) show an quench effect on the fluorescence intensity of the CNPs. The multicolor CNPs were successfully used as fluorescent probes for mouse melanoma cells imaging. The results indicate that the multicolor CNPs derived from cellulose and cyclodextrin may have a great potential for the applications in bioimaging.


Pretreatment serum C-reactive protein is a significant prognostic factor in patients with soft tissue metastases.
Soft tissue metastasis is rarer than bone metastasis. Patients with soft tissue metastasis generally have a dismal prognosis. The treatment for metastatic lesions is sometimes difficult, because the prognostic factors of patients with soft tissue metastasis remain unelucidated. Therefore, this study aimed to identify these prognostic factors. Thirty-one patients with soft tissue metastasis were included in the study. We evaluated associations of overall survival with clinical parameters and inflammatory markers using Kaplan-Meier curves and Cox proportional hazards models. Twelve patients received surgery for soft tissue metastasis, while radiation therapy was performed in six cases. The median overall survival after the detection of soft tissue metastasis was 11 months. Univariate analysis revealed that detection of soft tissue metastasis after the multidisciplinary treatment (P = 0.01); solitary metastasis (P = 0.0003); and pretreatment C-reactive protein (CRP) level < 0.4 mg/dL (P < 0.0001), white blood cell count < 8500 × 103/μL (P = 0.0003), and neutrophil-to-lymphocyte ratio < 5 (P = 0.02) were significant good prognostic factors. Multivariate analysis revealed that a CRP value < 0.4 mg/dL (P = 0.07) and solitary metastasis (P = 0.09) were possible significant predictors of survival. Furthermore, in case of CRP levels <0.4 mg/dL and metastatic tumor resection, patients had a good prognosis; however, when the CRP levels increased to 0.4 mg/dL and above, patients had a poor prognosis, irrespective of tumor resection. CRP is potentially useful for determining the indication of radical metastasectomy in soft tissue metastasis.


Lethal neonatal deficiency of carnitine palmitoyltransferase II associated with dysgenesis of the brain and kidneys.
We describe neonatal onset of a lethal multiorgan deficiency of carnitine palmitoyltransferase II (CPT II) associated with dysmorphic features, cardiomyopathy, and cystic dysplasia of the brain and kidneys. Concentrations of long-chain acylcarnitines were evaluated in blood and multiple tissues, diffuse lipid accumulation was present at autopsy, and a profound deficiency of CPT II activity was evident in heart, liver, muscle, and kidney tissue. This disorder constitutes another recognizable malformation syndrome with a metabolic basis. Deficiency of CPT II should be included in the differential diagnosis of patients with cystic renal dysplasia, dysmorphism, central nervous system malformations, and early death, along with glutaric acidemia type II, Zellweger syndrome, and other disorders in which peroxisomal beta-oxidation is impaired. The clinicopathologic similarities among these disorders raise the possibility that a common biochemical mechanism, namely the disruption of beta-oxidation of fatty acids, is responsible for the abnormal organogenesis.


Maternal and early postnatal immune activation produce sex-specific effects on autism-like behaviors and neuroimmune function in mice.
Increasing evidence suggests a role for inflammation in neuropsychiatric conditions including autism spectrum disorder (ASD), a neurodevelopmental syndrome with higher prevalence in males than females. Here we examined the effects of early-life immune system activation (EIA)-comprising regimens of prenatal, early postnatal, or combined ("two-hit") immune activation-on the core behavioral features of ASD (decreased social interaction, increased repetitive behavior, and aberrant communication) in C57BL/6J mice. We treated timed-pregnant mice with polyinosinic:polycytidylic acid (Poly I:C) on gestational day 12.5 to produce maternal immune activation (MIA). Some offspring also received lipopolysaccharide (LPS) on postnatal day 9 to produce postnatal immune activation (PIA). EIA produced disruptions in social behavior and increases in repetitive behaviors that were larger in males than in females. Ultrasonic vocalizations (USVs) were altered in both sexes. Molecular studies revealed that EIA also produced prominent sex-specific changes in inflammation-related gene expression in the brain. Whereas both sexes showed increases in pro-inflammatory factors, as reflected by levels of mRNA and protein, expression of anti-inflammatory factors was decreased in males but increased in females. Our findings demonstrate that EIA can produce sex-specific behavioral effects and immune responses in the brain, and identify molecular processes that may contribute to resilience in females.


Regional variation in thyroid cancer incidence in Belgium is associated with variation in thyroid imaging and thyroid disease management.
Increased thyroid cancer incidence is at least partially attributed to increased detection and shows considerable regional variation. We investigated whether regional variation in cancer incidence was associated with variations in thyroid disease management. We conducted a retrospective population-based cohort study that involved linking data from the Belgian Health Insurance database and the Belgian Cancer Registry to compare thyroid-related procedures between regions with high and low cancer incidence. Primary outcome measures were rates of TSH testing, imaging, fine-needle aspiration cytology (FNAC), and thyroid surgery. Secondary study outcomes were proportions of subjects with thyrotoxicosis and nodular disease treated with surgery, of subjects treated with surgery preceded by FNAC or with synchronous lymph node dissection, and of thyroid cancer diagnosis after surgery. The rate of TSH testing was similar, but the rate of imaging was lower in the low incidence region. The rate of FNAC was similar, whereas the rate of surgery was lower in the low incidence region (34 [95% CI 33; 35 ] vs 80 [95% CI 79; 81 ] per 100,000 person years in the high incidence region; P < .05). In the low incidence region compared to the high incidence region, surgery represented a less chosen therapy for euthyroid nodular disease patients (47% [95% CI 46; 48] vs 69% [95% CI 68; 70]; P < .05), proportionally more surgery was preceded by FNAC, more cancer was diagnosed after total thyroidectomy, and thyroid cancer patients had more preoperative FNAC and synchronous lymph node dissection. Regional variation in thyroid cancer incidence, most marked for low-risk disease, is associated with different usage of thyroid imaging and surgery, supporting variable detection as a key determinant in geographic variation.


Substance-induced psychosis as a risk factor for unipolar depression or anxiety disorders-A nationwide register-based prospective cohort study.
Substance-induced psychosis has previously been linked to increased incidence of schizophrenia and bipolar disorder. We aimed to investigate if substance-induced psychosis is associated with increased risk of depression or anxiety. We conducted a nationwide prospective register-based cohort study from 1994 to 2017, including all individuals with substance-induced psychosis, and age-and-sex matched controls without substance-induced psychosis. We investigated time to either depression or anxiety, as well as time to depression and time to anxiety, in stratified Cox regression models. We included 5,557 individuals with substance-induced psychosis and 55,562 controls. Substance-induced psychosis was associated with increased risk of either depression or anxiety (HR=7.05, 95% CI 6.71-7.41), depression (HR=5.40, 95% CI 4.77-6.11), or anxiety (HR=7.05, 95% CI 5.99-8.31). Analyses of individual types of substance-induced psychosis revealed similar hazard ratios across substances. Associations between substance-induced psychosis and depression or anxiety were stronger in people without preceding alcohol or substance use disorders. While strongest shortly after incident substance-induced psychosis, the increased incidence of depression and anxiety remained more than double over the full period of follow-up. Only psychiatric disorders treated either in psychiatric inpatient or outpatient units, supplemented with information on psychiatric medication, was available. Exact times of onset were similarly unknown, and only dates of first treatment were available. Substance-induced psychosis is a strong predictor of later onset of depression or anxiety. Regardless of whether this association is causal, this highlights the need for increased monitoring and possibly improved treatment of patients with substance-induced psychosis.


Quality of life in GERD patients: medical treatment versus antireflux surgery.
Medical and surgical treatments are able to improve symptoms in patients with gastroesophageal reflux disease (GERD). The aim of this study was to evaluate the outcome in GERD patients without therapy, under continuous medical treatment, and after laparoscopic antireflux surgery. Five hundred seventy-nine consecutive patients underwent medical or surgical treatment for GERD-induced symptoms. Patients were studied in detail before and after treatment by means of a symptom questionnaire, endoscopy, esophageal manometry, 24-hour esophageal pH monitoring, and a barium esophagogram. In addition, quality of life was measured by the means of the Gastrointestinal Quality of Life Index (GIQLI) and the Health-Related Quality of Life (HRQL) questionnaire. Surgery was indicated and performed in 351 patients with persistent or recurrent GERD symptoms and/or complications, and in patients preferring surgery to medical treatment, despite the use of an adequate medication. The remaining 228 patients were treated with proton pump inhibitors (PPI) in the standard dose, or if required, the double dose. The outcome was assessed 3 and 12 months after treatment. While symptoms and quality of life were highly impaired in GERD patients without therapy compared with normal people, a significant improvement was obtained by PPI therapy. Following surgery, quality of life was normalized in all subsections and was significantly higher compared with the medically treated group. These results stayed constant in short-term and intermediate follow-up. Medical and surgical therapies are both able to improve symptoms and quality of life in GERD patients. Nevertheless, the outcome is significantly better following surgery. It can be suggested that surgical treatment may be the more successful therapy in the long-term.


Paris polyphylla extract attenuates colitis in mice by regulating PPAR-γ mediated Treg/Th17 balance.
Paris polyphylla Sm. (P.P), is a widely-used traditional Chinese medicine (TCM) in the treatment of wound, throat sores and snakebites. Furthermore, P.P was recorded as an anti-inflammatory drug by the Chinese Pharmacopoeia. We sought to decipher the anti-inflammatory effect of P.P on ulcerative colitis (UC); specifically, to explore whether P.P attenuates colitis by restoring the regulatory T cells (Tregs) and T helper 17 (Th17) cells balance and its mechanism. We treated experimental colitis mice with extracts of Paris polyphylla (EPP). The percentage of Tregs and Th17 cells were measured using flow cytometry, and their secreted cytokines levels were evaluated employing ELISA. The expression of peroxisome proliferator-activated receptor gamma (PPAR-γ) in colon tissues was detected using immunofluorescence. Furthermore, GW9662, a PPAR-γ antagonist, was used to validate the mechanism of EPP in restoring the Treg/Th17 balance. The EPP effectively alleviated the clinical symptoms and inflammatory cytokine levels in mice with colitis. EPP treatment also restored the impaired Treg/Th17 balance in mice. Furthermore, EPP treatment promoted PPAR-γ expression and reduced HIF-1α and p-STAT3 expression in colon tissues, whereas PPAR-γ inhibition blocked the effects of EPP in mice models. Our study indicates that EPP exhibit excellent anti-inflammatory properties via restoring PPAR-γ/STAT3/HIF-1α axis-mediated Treg/Th17 balance in colitis mice. Hence, P. polyphylla is a promising medicinal plant-based alternative for managing colitis that requires further clinical validation.


Design, development and demonstration of an improved bird washing machine.
Since oil was first extracted, pollution of the seas and oceans or adjacent coasts has been an obstacle for the oil industry and environmental activists. The major concern is oil discharge into the water which may lead to birds' affliction or death, besides putting marine life in jeopardy. This paper presents the first description of the design and implementation of a new bird washing machine that can be utilized for cleaning of oil-coated birds with the minimum of stress. The machine is equipped with a pneumatic system comprised of 19 moving nozzles which evenly cover the bird's body and is designed to be used in contaminated environments where a vast number of birds are affected. Experimental trials show an improvement in operation efficiency compared to other methods in a reduction in washing time, energy consumption and a decrease in fatality rate of washed birds.


Annexin A9 promotes cell proliferation by regulating the Wnt signaling pathway in colorectal cancer.
Colorectal cancer (CRC) is one of the leading causes of cancer-related mortality worldwide. Expression of Annexin A9 (ANXA9), a member of the annexin A family, is upregulated in CRC. However, the molecular role of ANXA9 in CRC remains unknown. In the present study, we aimed to investigate the function of ANXA9 and to elucidate the mechanisms underlying its regulation in CRC. In this study, mRNA expression data and clinical information were downloaded from The Cancer Genome Atlas (TCGA) and GEPIA database, respectively. Kaplan-Meier analysis was used to analyze the survival rates. LinkedOmics and Metascape databases were used to explore the potential mechanisms of regulation of ANXA9 and to identify genes co-expressed with ANXA9. Finally, in vitro experiments were used to evaluate the function of ANXA9 and explore potential mechanisms. We found that ANXA9 expression was significantly elevated in CRC tissue and cells. High ANXA9 expression was associated with shorter overall survival, poorer disease specific survival, as well as with patient age, clinical stage, M stage, and OS events in CRC. Knockdown of ANXA9 inhibited cell proliferation, invasion, migratory potential, and cell cycle arrest. Mechanistically, functional analysis revealed that genes co-expressed with ANXA9 were mainly enriched in the Wnt signaling pathway. ANXA9 deletion suppressed cell proliferation via the Wnt signaling pathway, while Wnt activation reversed the effects of ANXA9. In conclusion, ANXA9 may promote CRC progression by regulating the Wnt signaling pathway and may be a potential diagnostic biomarker in the clinical management of CRC.


Oral-motor function in older people: Exploring new dimensions of frailty.
Oral-motor performance for speech and swallowing, and verbal fluency represent important domains that can determine frailty thresholds in older people. The study aims to explore the association between oral-motor performance and verbal fluency to achieve a comprehensive measurement of the frailty phenotype. An exploratory and inferential cross-sectional study was carried out in two nursing homes and 2 day care centres. The study comprised a sample of 95 individuals with a mean age of 83 years. The oral-diadochokinesis test (ODDK), water swallow test (WST), time of mastication and swallowing (TOMASS), maximum tongue pressure, verbal fluency, physical phenotype and Mini-Mental State Examination (MMSE) were used as measurements variables. The comparison of mean values between the performance of observed and normative values for the target population was shown to be statistically significant for all the measurements and between the pre-frail and frail. The results of this study add to the growing body of evidence that oral-motor function for speech and swallowing and verbal fluency are relevant to the diagnosis and management of the frailty condition in older people. The frailty syndrome is more than just the traditionally observed physical function, rather it is a multidimensional construct, where additional dimensions should be considered when evaluating frailty in clinical practice.


Transcystoscopic intracavitary irradiation for carcinoma of the bladder: technique and preliminary clinical results.
A remote controlled transcystoscopic intracavitary after-loading unit is introduced for irradiation therapy of bladder carcinoma. With intense radiation therapy a significant dose can be delivered to the tumor during a short interval. We treated 12 patients with transitional cell carcinoma of the bladder using transcystoscopic intracavitary irradiation. Of the patients 9 initially had a complete response, although within several months 3 subsequently had recurrence in a different part of the bladder. Technical difficulties and severe complications were not encountered. The preliminary results and technique of transcystoscopic intracavitary irradiation are reported.


ω-Alkynyl arachidonic acid promotes anti-inflammatory macrophage M2 polarization against acute myocardial infarction via regulating the cross-talk between PKM2, HIF-1α and iNOS.
Macrophage polarization determines the timing for the switch from the inflammation phase to the inflammation resolution phase after acute myocardial infarction. The aim of the present study was to investigate whether ω-alkynyl arachidonic acid could mitigate the inflammatory lipid mediators in the regulation of macrophage phenotypes and functions with a special regard to myocardial infarction. We initially discovered that ω-alkynyl arachidonic acid selectively suppressed the up-regulation of inducible nitric oxide synthase (iNOS) over cyclooxygenase-2 (COX-2) in LPS-stimulated macrophages. ω-Alkynyl arachidonic acid also reduced the expression of macrophage M1 biomarkers (e.g., TNF-α, CXCL10, iNOS and IL-6) but increased the expression of macrophage M2 biomarkers (e.g., IL-10 and arginase-1) in LPS-stimulated macrophages. Moreover, ω-alkynyl arachidonic acid markedly enhanced the phagocytotic activity of macrophages against fluorescently-labeled beads or apoptotic H9c2 cardiac cells. We further investigated the in vivo cardioprotective activities of ω-alkynyl arachidonic acid in a mouse model of myocardial infarction. ω-Alkynyl arachidonic acid indeed reduced infarct size, cardiac damage and the leakage of myocardial enzymes CK-MB. Mechanistic studies revealed that ω-alkynyl arachidonic acid suppressed the overexpression and nuclear translocation of glycolytic enzyme PKM2 in LPS-stimulated macrophages. Furthermore, co-immunoprecipitation assay suggested that ω-alkynyl arachidonic acid disrupted the interaction between PKM2 and HIF-1α. Consequently, ω-alkynyl arachidonic acid diminished HIF-1α binding to the HRE sequence in iNOS promoter in response to LPS stimulation. Collectively, ω-alkynyl arachidonic acid may promote the anti-inflammatory M2 polarization of macrophages in acute myocardial infarction via regulating the cross-talk between PKM2, HIF-1α and iNOS.


It Takes a Village to Raise a Child: Understanding and Expanding the Concept of the "Village".
This perspective article defines and discusses the concept of the "village" when working with families who are experiencing multiple adversities. The article starts with a discussion on what is meant generally by a village approach, followed by a historical overview of how families living in adversity have been defined and positioned. The need to move past a siloed, professional centric approach when working with families is then presented. Using a model of social connections, based on Bronfenbrenner's ecological theory, we then identify who the "villagers" might be. Some potential principles for how the village might work with families living with adversity are presented, along with two case studies, to demonstrate how these principles might be enacted. This perspective article provides an overview and discussion of "the village" concept, rather than present a definitive set of guidelines or recommendations.


Structure of aldose reductase from Giardia lamblia.
Giardia lamblia is an anaerobic aerotolerant eukaryotic parasite of the intestines. It is believed to have diverged early from eukarya during evolution and is thus lacking in many of the typical eukaryotic organelles and biochemical pathways. Most conspicuously, mitochondria and the associated machinery of oxidative phosphorylation are absent; instead, energy is derived from substrate-level phosphorylation. Here, the 1.75 Å resolution crystal structure of G. lamblia aldose reductase heterologously expressed in Escherichia coli is reported. As in other oxidoreductases, G. lamblia aldose reductase adopts a TIM-barrel conformation with the NADP(+)-binding site located within the eight β-strands of the interior.


[Multi-organ involvement in recurrent Moschcowitz disease. Splenectomy as ultima ratio in a therapeutic dilemma?].
The symptoms primarily described in Morbus Moschcowitz included thrombocytopenic purpura and hemolytic anemia. In addition it presents with a variety of clinical manifestations depending on the organs involved (i.e. neurological, renal, gastrointestinal, cardiac involvement). It is a rare disease and the pathogenesis still remains unclear. The efficacy of derived therapeutical concepts can hardly be assessed in controlled trials. Currently the main option seems to be plasma therapy. In non-responders surgical procedures (splenectomy) may be of benefit to the patient.


Novel AKT1-GLI3-VMP1 pathway mediates KRAS oncogene-induced autophagy in cancer cells.
Autophagy is an evolutionarily conserved degradation process of cytoplasmic cellular constituents. It has been suggested that autophagy plays a role in tumor promotion and progression downstream oncogenic pathways; however, the molecular mechanisms underlying this phenomenon have not been elucidated. Here, we provide both in vitro and in vivo evidence of a novel signaling pathway whereby the oncogene KRAS induces the expression of VMP1, a molecule needed for the formation of the authophagosome and capable of inducing autophagy, even under nutrient-replete conditions. RNAi experiments demonstrated that KRAS requires VMP1 to induce autophagy. Analysis of the mechanisms identified GLI3, a transcription factor regulated by the Hedgehog pathway, as an effector of KRAS signaling. GLI3 regulates autophagy as well as the expression and promoter activity of VMP1 in a Hedgehog-independent manner. Chromatin immunoprecipitation assays demonstrated that GLI3 binds to the VMP1 promoter and complexes with the histone acetyltransferase p300 to regulate promoter activity. Knockdown of p300 impaired KRAS- and GLI3-induced activation of this promoter. Finally, we identified the PI3K-AKT1 pathway as the signaling pathway mediating the expression and promoter activity of VMP1 upstream of the GLI3-p300 complex. Together, these data provide evidence of a new regulatory mechanism involved in autophagy that integrates this cellular process into the molecular network of events regulating oncogene-induced autophagy.


Stimulus estimation and the overprediction of fear: a comment on two studies.
The overprediction of fear is the tendency to overestimate the amount of fear one will experience in a subjectively threatening situation. This bias is thought to promote excessive avoidance and limit the opportunity for corrective learning. The stimulus estimation model states that the overprediction of fear arises from the overprediction of the danger features of the stimulus and the underprediction of available safety resources. Two recent studies reportedly failed to support the model (Arntz, Hildebrande & van den Hout, Behaviour Research and Therapy, 32, 709-722, 1994; Telch, Valentiner & Bolte, Behaviour Research and Therapy, 32, 747-751, 1994). The present commentary shows that both studies suffered significant methodological problems and neither can be said to have tested the model. Criteria for evaluating the model are briefly discussed.


Changes in the swimming activity and the glutathione S-transferase activity of Jenynsia multidentata fed with microcystin-RR.
We report the effects of sublethal doses of microcystin-RR (MC-RR) on the swimming activity of Jenynsia multidentata as well as the simultaneous response of its detoxication system by measuring glutathion S-transferase (GST) activities in the liver and brain of fish. MC-RR was applied on the food pellets at doses of 0.01, 0.1 and 1 microg g(-1). Swimming activity was recorded 10 min each hour over 24h by using a computer-based image processing system, which facilitates quantification of two measures of fish swimming behaviour (average velocity, movement percentage). Results show that low levels of cyanotoxin increased the swimming activity, while the highest dose used produced significant changes with respect to control group only since approximately 20 h of exposure, when the swimming activity was decreased. On the other hand, GST activity was significantly increased only in the liver and brain of fish fed with the highest MC-RR dose. Both results suggest that fish are reacting to the stress caused by low doses of MC-RR by increasing their swimming activity, raising further questions on the probable neurotoxicity of MCs, and presenting the behavioural change as a good biomarker of early toxic stress. On the other hand, fish reduced their swimming speed at the highest MC-RR dose, when the detoxication activity began, which can be hypothesized to be a reallocation of their energy, favouring detoxication over swimming activity.


Relationship between inflammatory markers and coronary slow flow in type 2 diabetic patients.
Diabetes is a serious and quickly expanding global health problem. Cardiovascular disease is the leading cause of mortality in type 2 diabetes mellitus (T2DM) patients. Coronary slow flow (CSF) is characterised by delayed distal perfusion during coronary angiography with normal coronary arteries. This study aimed to investigate the correlation between CSF and inflammatory markers regarding glycemic status in T2DM. This cross-sectional study included 120 patients who were divided equally into 4 groups according to their glycemic control and presence or absence of coronary slow flow: Group I included patients with T2DM with good glycemic control without CSF; Group II included patients with T2DM with good glycemic control and CSF; Group III included patients with T2DM with poor glycemic control without CSF; and Group IV included patients with T2DM with poor glycemic control and CSF. The neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), C-reactive protein (CRP), platelets, hematocrit, and haemoglobin were also evaluated as risk factors for coronary slow flow. This study showed that body mass index (BMI), hematocrit level, NLR, and CRP demonstrated a moderate but significant correlation (r = 0.53) with CSF in poorly controlled T2DM. NLR cutoff > 2.1 could predict CSF in poorly controlled T2DM with a modest sensitivity and specificity. A 1.9 increase in HbA1c increases the likelihood of coronary slow flow. Dylipidemia increases the likelihood of coronary slow flow by 0.18 times. Other predictors for coronary slow flow include NLR, PLR, CRP, platelets, hematocrit, and hemoglobin. The effect of the predictors is still statistically significant after being adjusted for glycemic status, age, and sex (p < 0.001). Poor glycemic control increases the incidence of CSF. This supports the hypothesis that CSF is related to endothelial dysfunction as poor glycemic control causes endothelial dysfunction due to inflammation. ZU-IRB#9419-3-4-2022 Registered 3 April 2022, email. firstname.lastname@example.com .


Molecular and in situ characterization of cadmium-resistant diversified extremophilic strains of Pseudomonas for their bioremediation potential.
Cadmium-resistant strains psychrotolerant Pseudomonas putida SB32 and alkalophilic Pseudomonas monteilli SB35 were originally isolated from the soil of Semera mines, Palamau, Jharkhand, India. Further, to unravel the mechanism involved in cadmium resistance, plasmid DNA was isolated from the strains and subjected to amplification of the czc gene, which is responsible for the efflux of three metal cations, viz. Co, Zn and Cd, from the cell. Furthermore, the amplicon was cloned into pDrive cloning vector and sequenced. When compared with the available database, the sequence homology of the cloned gene showed the presence of a partial czcA gene sequence, thereby indicating the presence of a plasmid-mediated efflux mechanism for resistance in both strains. These results were further confirmed by atomic absorption spectroscopy and transmission electron microscopy. Moreover, the strains were characterized functionally for their bioremediation potential in cadmium-contaminated soil by performing an in situ experiment using soybean plant. A marked increase in agronomical parameters was observed in presence of both strains. Further, the concentration of metal ions decreased in both plants and soil in the presence of these bioinoculants.


Assessment of factors affecting vaccine cold chain management practice in public health institutions in east Gojam zone of Amhara region.
Maintaining quality of vaccines is one of the main challenges of immunization programs in Ethiopia. The objective of this study is to assess the factor affecting vaccine cold chain management practice in immunization health institutions in East Gojam zone of Amhara region, Ethiopia. An institutional based cross-sectional study was conducted from March to April 2017 in ten districts of East Gojam zone of Amhara Region. Descriptive statistics and Logistic regression analysis were carried out to identify factors related to the practice of cold chain management. Among 60 health institutions, only 46(76.7%) had functional refrigerators. Twenty-one (35%) had a functional generator for backup service and 28(46.6%) had a car/motorbike for transportation of vaccines in case of refrigerator/power failure. Twenty-nine (48.3%) had known the correct vaccine storage temperature (2 °C - 8 °C) in the refrigerator and the results of this study revealed that only 23(38.3%) of respondents had sufficient knowledge about vaccine cold chain management. The finding of this study also revealed that 35(58.3%) had appropriate vaccine cold chain management practice and the rest 25(41.7%) had inappropriate practice. Logistic regression showed us the knowledge gap and profession were significantly associated with vaccine cold chain management practice at P < 0.05. This study indicates that there was a knowledge gap of health workers who are working on cold chain management. There is an urgent need to improve knowledge and practice on cold chain management through improved supervision and training at a different level of health care system.


The meanings of r- and K-selection.
This paper catalogues several different dichotomies that have all been termed r- and K-selection. The status of the concept of r- and K-selection is discussed and a more restricted usage of the terms is recommended.


Improvement in glucose metabolism in adult male offspring of maternal mice fed diets supplemented with inulin via regulation of the hepatic long noncoding RNA profile.
Maternal overnutrition during pregnancy and lactation is an important risk factor for the later development of metabolic disease, especially diabetes, among mothers and their offspring. As a fructan-type plant polysaccharide, inulin has prebiotic functions and is widely used as a natural antidiabetic supplement. However, to date, the mechanism of maternal inulin treatment in the livers of offspring has not been addressed, especially with respect to long noncoding RNAs (lncRNAs). In this study, female C57BL6/J mice were fed either a high-fat diet (HFD) with or without inulin supplementation or a standard rodent diet (SD) during gestation and lactation. After the offspring were weaned, they were fed a SD for 5 weeks. At 8 weeks of age, the glucose metabolism indexes of the offspring were assessed, and their livers were collected to assay lncRNA and mRNA profiles to investigate the effects of early maternal inulin intervention on offspring. Our results indicate that male offspring from HFD-fed dams displayed glucose intolerance and an insulin resistance phenotype at 8 weeks of age. Early maternal inulin intervention improved glucose metabolism in male offspring of mothers fed a HFD during gestation and lactation. The lncRNA and mRNA profile data revealed that compared with the offspring from HFD dams, offspring from the early inulin intervention dams had 99 differentially expressed hepatic lncRNAs and 529 differentially expressed mRNAs. The differentially expressed lncRNA-mRNA coexpression analysis demonstrated that early maternal inulin intervention may change hepatic lncRNA expression in offspring; there lncRNAs are involved in metabolic pathways and the AMP-activated protein kinase signaling pathway. Importantly, the early maternal inulin intervention alleviated glucose metabolism by inhibiting the lncRNA Serpina4-ps1/let-7b-5p/Ppargc1a as a competing endogenous RNA in male offspring.


Studies on the association of the Epstein-Barr virus genome and human chromosomes.
We have used human-mouse somatic cell hybrid to study the association between the EBV genome and the cellular genome. Attempts were made to identify a specific human chromosome(s) with which the EBV genome is associated. Our data suggest that at least in the mouse fibroblast/Burkitt lymphoma hybrid combination studies, the EBV genome is not associated with any specific human chromosome.


Strategy to Design a Flexible and Macromolecular Sensor to Bind Cd2+ Ions: A Complete Photophysical Analysis and Bio-Imaging Study.
A novel triazole-bridged coumarin-benzimidazole-conjugated fluorescence sensor (4) has been developed for selective detection of Cd2+ over other competitive metal ions. The sensor exhibited quick "turn-on" responses upon interaction with a very low level of Cd2+ (14 nM). The photophysical changes in the complexation of Cd2+ with sensor 4 have been explained through the excited-state intramolecular proton transfer mechanism. The involvement of benzimidazole and triazole moieties in Cd2+ binding was confirmed by different spectroscopic techniques such as UV-vis, Fourier transform infrared, nuclear magnetic resonance, and ESI mass. The diameter of the circular shape of the sensor decreased upon complexation with Cd2+, which was confirmed by field-emission scanning electron microscopy. Furthermore, the quantum chemical (density functional theory) calculation supported the mechanism of interactions and the mode of binding of 4 toward Cd2+. The sensor was more effective for finding Cd2+ in two living cells, C6 (rat glial cell) and Hep G2 (human liver cell).


[Surgical treatment experience of different approaches in eight cases with parapharyngeal space foreign bodies].
To analyze the surgical treatment of parapharyngeal space foreign body and to discuss feasibility of Endoscopic transoral approach. Reviewing surgical treatment in eight cases of parapharyngeal space foreign body, nature and location of foreign body. surgical methods, and treatment results were analyzed. Eight patients' foreign bodies were all removed by surgical treatment, there were no postoperative infection. Three cases of foreign body were removed through laterocervical approaches, one removed through submandibular approach. In four cases the foreign body were removed through Endoscopy, two of them were transoral approach and the other two were taken out after antiotomy. Different approaches should be taken in the surgery treatment of parapharyngeal space foreign body according to imaging positioning.


Defibrotide reduces infarct size in a rabbit model of experimental myocardial ischaemia and reperfusion.
1. Defibrotide, a single-stranded polydeoxyribonucleotide obtained from bovine lungs, has significant anti-thrombotic, pro-fibrinolytic and prostacyclin-stimulating properties. 2. The present study was designed to evaluate the effects of defibrotide on infarct size and regional myocardial blood flow in a rabbit model of myocardial ischaemia and reperfusion. 3. Defibrotide (32 mg kg-1 bolus + 32 mg kg-1 h-1, i.v.) either with or without co-administration of indomethacin (5 mg kg-1 x 2, i.v.) was administered 5 min after occlusion of the left anterior-lateral coronary artery and continued during the 60 min occlusion and subsequent 3 h reperfusion periods. 4. Defibrotide significantly attenuated the ischaemia-induced ST-segment elevation and abolished the reperfusion-related changes (R-wave reduction and Q-wave development) in the electrocardiogram. In addition, defibrotide significantly improved myocardial blood flow in normal and in ischaemic, but not in infarcted sections of the heart. The improvement in blood flow in normal perfused myocardium, but not in the ischaemic area was prevented by indomethacin. 5. Although the area at risk was similar in all animal groups studied, defibrotide treatment resulted in a 51% reduction of infarct size. Indomethacin treatment abolished the reduction of infarct size seen with defibrotide alone. 6. The data demonstrate a considerable cardioprotective effect of defibrotide in the reperfused ischaemic rabbit myocardium. This effect may be related, at least in part, to a stimulation of endogenous prostaglandin formation. Other possible mechanisms are discussed.


Exploring the Intersection Between Academic and Professional Practice During the COVID-19 Pandemic: Undergraduate and Graduate Nursing Students' Experiences.
The coronavirus disease-2019 (COVID-19) pandemic has implications for students who are also nurses. This qualitative descriptive study used a practice development approach to explore the intersection between academic and professional work experiences for undergraduate Post-Diploma Registered Practical Nurses bridging to Registered Nurse Bachelor of Science in Nursing students and Master of Nursing graduate nursing students during the first wave of the COVID-19 pandemic. The study incorporated critical aesthetic reflections that focused on the personal and aesthetic ways of knowing, as a data collection approach and knowledge dissemination strategy. Analysis of the narrative component of participants' reflections revealed the following themes: sensing a "call to duty," experiencing a myriad of emotions, shifting societal and individual perceptions of nursing, and learning in an uncertain environment. The results of the study can inform educational strategies and academic policies to support this unique nursing population, who are frontline practitioners as well as student learners.


Non-frozen transports of whole blood samples do not cause relevant bias for global coagulation tests in clinical trials evaluating the drug safety.
Sample shipments with dry ice have a large economic impact on clinical research. Therefore, the bias caused for global coagulation tests by non-frozen transports of whole blood instead of frozen plasma was investigated experimentally and by a meta-analysis of 6-year central laboratory data. In the experiment, aliquots from 14 healthy volunteers were kept as whole blood at 20+/-2 degrees C and as frozen plasma until an analysis of prothrombin time (PT), activated partial thromboplastin time (aPTT), thrombin time (TT), and antithrombin III (ATIII) at day 0, 1, 2, and 3 from collection. Within these 3 days only PT and aPTT demonstrated any changes: in blood samples kept at 20+/-2 degrees C these amounted about 10% for both. In frozen plasma, aPTT did not change whereas PT increased by 14%. In a meta-analysis of central laboratory data, PT and aPTT results were grouped across various phase II-IV trials by the type of sample transfer, either as frozen plasma on dry ice or non-frozen as whole blood. For the latter the mean difference to a reference group of phase I trials with same-day analysis was in line with the amount of bias found in the experiment (aPTT, 34.6+/-6.0 vs. 31.6+/-3.5 s; PT, 87.7+/-13.3 vs. 97.3+/-7.9%). The consistent bias resulted in shifted, but still normal distribution curves with a total rate of clinically relevant outliers of about 1.9% for aPTT and 2.4% for PT. Biases thus appear irrelevant for a common safety evaluation within clinical trials. Non-frozen whole blood transports for the measurement of global coagulation tests appear justified for this purpose, if protocols do not require frozen shipments for other reasons. However, transit time must not exceed 2 days and pre-analytical conditions should be consistent within the same trial.


The development of the septal region in the rat. I. Neurogenesis examined with 3H-thymidine autoradiography.
Neurogenesis in the rat septal region was examined with 3H-thymidine autoradiography. The rats in the prenatal groups were the offspring of pregnant females given two injections of 3H-thymidine on consecutive days in an overlapping series: embryonic day (E) 13 + E14, E14 + E15,. . . E21 + E22. The rats in the postnatal groups were injected in a nonoverlapping series: the day of birth and postnatal day (P) 1, P2 + P3, P3 + P4. On 60 days of age, the percentage of labelled cells and the proportion of cells added during each day of formation were determined at several anatomical levels within the midline nuclear group (nucleus of the diagonal band, medial and triangular septal nuclei), the lateral septal nucleus, and the ventrolateral nuclear group (nucleus accumbens, bed nuclei of the stria terminalis and the anterior commissure). The neurons within each nuclear group form in significantly different waves, those of the midline group forming between E13-E17, the lateral septal nucleus between E15-E19, the bed nuclei of the stria terminalis and anterior commissure between E14-E18, the nucleus accumbens between E17-P2. All nuclei and nuclear groups show characteristic gradients of formation. Both the midline nuclear group and the bed nucleus of the stria terminalis (including the commissural bed nucleus) have their earliest forming neurons lying near the crossing of the anterior commissure; younger nuerons are located both rostrally and caudally with the youngest neurons lying in the most rostral extension of the diagonal band nucleus and the strial bed nucleus. The lateral septal nucleus forms along a strong mediolateral gradient throughout its length after neurogenesis is almost complete in the midline nuclear group. Throughout the length of the nucleus accumbens, the oldest neurons are located ventrally while progressively younger cells are found dorsally beneath the inferior horn of the lateral ventricle.


Lysine-Specific Demethylase 4D Is Critical for the Regulation of the Cell Cycle and Antioxidant Capacity in Goat Fibroblast Cells.
Oxidative damage to skin fibroblast cells is a causative factor in many skin diseases. Previous studies have reported that lysine-specific demethylase 4D (Kdm4d) is involved in DNA replication, but its role on antioxidant capacity remains unclear. In the present study, we used goat fibroblast cells (GFCs) as the research model and identified 504 up-regulated and 1013 down-regulated genes following the knockdown of Kdm4d, respectively. The down-regulated genes of this enzyme were found to be enriched in the cell cycle, DNA replication, mitotic processes, and the oxidative phosphorylation pathway, as previously revealed from gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG), and gene set enrichment analysis (GSEA), suggesting vital roles of the Kdm4d enzyme in the cell cycle and in antioxidant regulation. To this end, we found the cell proliferation rate was significantly decreased after the knockdown of Kdm4d. Moreover, both the mRNA and protein expression levels of superoxide dismutase 2 (SOD2), one of the major antioxidant enzymes, was decreased, while the reactive oxygen species (ROS) level was significantly increased in Kdm4d knocked-down cells. In addition, the expression of γH2A histone family member X (γH2AX) increased significantly, indicating the presence of DNA double-strand breaks after the knockdown of the Kdm4d enzyme. In conclusion, the knockdown of Kdm4d inhibited DNA replication and the cell cycle, repressed the expression of SOD2, and increased the generation of ROS, which led to the production of DNA damage in GFCs. Our data will be helpful for understanding the mechanism underlying antioxidant capacity regulation in fibroblast cells.


Laser flash spectroscopy of methylene blue with nucleic acids. Effects of ionic strength and pH.
Laser flash spectroscopic measurements were made on methylene blue complexed to the synthetic polynucleotides poly[d(G-C)] and poly[d(A-T)] in solutions of varying ionic strength and pH. Triplet decay rates and rates of triplet quenching by oxygen have been measured for the polymer/dye solutions. The triplet decay and oxygen quenching rates of methylene blue in complex with poly[d(A-T)] are much less sensitive than those with poly[d(G-C)] with respect to variations in ionic strength. It is also shown that protonation of the triplet state of MB+ with poly[d(A-T)] is slower than that of free dye. These results indicate strong binding of the dye to poly[d(A-T)]. Excitation of the MB+/poly[d(A-T)] complex at 665 nm yielded single exponential decay kinetics, in contrast with excitation at 600 nm where double exponential kinetics were measured. This is tentatively assigned to excitation of dye dimer bound to this polymer.


Eril's Branch-Based Perforator Plus Chondro-Cutaneous Flap for Reconstruction of Upper Third Helico-Scaphal Defects: A Clinical and Cadaver Study.
Reconstruction of upper third helico-scaphal defects (HSDs) exacts the expertise of the reconstructive surgeon. These three-dimensional defects deviate from normal facial form and cause a significant impact on the psyche of the patients. The origin, course, and dimensions of the perforators of the Eril's branch arising from superficial temporal vessels (EBSTV) were delineated by dissection and injection study. They were examined in twenty-four adult specimens. A prospective cohort study was conducted from March 2017 to March 2020, including six patients whose upper third HSDs were reconstructed with perforator plus chondro cutaneous flap based on EBSTV. After the debridement of the defect, the perforator plus chondro cutaneous flap was harvested from retro auricular sulcus skin with attached cartilages from triangular fossa and interhelical groove. The donor site was closed primarily, and the flap was rotated onto the defect. Superficial temporal vessels give a constant Eril's branch at the level of the root of the helix (otobasion superioris), which in turn divides into two perforators -superficial (lateral) and deep (medial) perforators. All six patients with post-traumatic upper third HSDs (4 males and 2 females) after reconstruction were followed for an average period of 15 months. The average age was 36.5 yrs. On average, two cartilages of maximum dimensions 3 × 2.5 mm were included in the perforator plus chondro cutaneous flap based on the medial-deep perforator of the EBSTV. In all six cases, the flaps survived without any complications. The average final computed institutional objective and subjective score was 15.67 (excellent results) (chi-square test, the p value was (p = 0.035) significant). The perforator plus chondro-cutaneous flap based on deep medial perforator of EBSTV may be a useful addendum in the armamentarium of plastic surgery. This novel reconstruction technique of upper helical defects provides good shape, size, and aesthesis of the upper external ear.


[Ornithine decarboxylase activity in rat tissues in the late periods after irradiation].
Ornithine decarboxylase activity in some rat tissues was shown to grow at remote times after gamma irradiation (60Co) with nonlethal doses. The authors suggest that ornithine decarboxylase activity should be used as a biochemical marker of remote effects of ionizing radiation.


The conformational phase space of the photoprotective switch in the major light harvesting complex II.
Transitions between protein states are triggered by external stimuli. This knowledge leads to the control of protein function. Herein, we report a large scale (90 μs) study on the conformational space of the major light harvesting complex II, based on a comprehensive array of external stimuli.


Small peptides as modular catalysts for the direct asymmetric aldol reaction: ancient peptides with aldolase enzyme activity.
Simple peptides and their analogues having a primary amino group as the catalytic residue mediate the direct asymmetric intermolecular aldol reaction with high stereoselectivity and furnish the corresponding aldol products with up to 99% ee; this intrinsic ability of highly modular peptides may explain the initial molecular evolution of aldolase enzymes.


Preparative countercurrent chromatography with a slowly rotating helical tube.
The capability of a simple countercurrent chromatographic scheme for obtaining high-resolution preparative-scale separations was demonstrated with the separations of a series of dinitrophenyl (DNP) amino acids and peptides. Basic studies on stationary phase retention and partition efficiency with a low-viscosity chloroform-acetic acid-0.1 N hydrochloric acid (2:2:1) phase system together with the results previously obtained with a vascous n-butanol system suggest a general applicability of low-interfacial tension phase systems by the present method.


Migratory serpiginous corneal epitheliopathy (MSCE)- details of 4 cases.
To describe four cases of recurrent, fleeting corneal epithelial lesion, migratory serpiginous corneal epitheliopathy (MSCE). Description of cases, histopathology and discussion of treatment outcome. The study included 4 consecutive male patients; mean age was 21.25 years. Only one eye was affected. The common symptoms were irritation and tearing. Typical interpalpebral corneal lesion was a superficial greyish-white elevated corneal epithelium with stippled fluorescein staining in various geographic patterns. Corneal scrapings did not show any microorganisms and culture was negative; histopathology did not show corneal dysplasia. The lesions recurred within 1-3 weeks despite repeated scraping, application of a bandage contact lens, and a combination of topical antibiotics and lubricants. The recurrent lesion was of a different shape and eventually healed within 3-6 months without scarring. We describe a possibly new entity, unilateral migratory serpiginous corneal epitheliopathy (MSCE) affecting young males, with inconspicuous inflammation. Etiology is not known and usually takes several months to heal after repeated epithelial debridement.


Immune responses and safety of COVID-19 vaccination in atypical hemolytic uremic syndrome patients in Taiwan.
Atypical hemolytic uremic syndrome is a rare, life-threatening disorder which can be triggered by COVID 19 infection and COVID 19 vaccination then induce multiple organ failure. Our study is the first to evaluate immune responses to COVID-19 vaccination and safety in a cohort of patients in a local single-center study in Taiwan.. Results indicate that vaccines effectively shield aHUS patients from severe COVID-19 complications without significant safety concerns. A double booster dose for the third vaccine is essential for optimal efficacy. Anti-complement therapy did not influence vaccination effectiveness. Transplant aHUS patients had the lowest immune response titers, indicating a need for additional vaccine doses. Compared to healthcare workers, aHUS patients had poor T-cell responses. We noted a superior trend with mixed-type COVID-19 vaccinations in aHUS patients, while fixed-type mRNA demonstrated better results in healthcare workers. Our findings endorse COVID-19 vaccination as a potent strategy to safeguard aHUS patients from severe complications, emphasizing the importance of vigilant monitoring pre- and post-vaccination.


Analysis of quinolone antibiotic derivatives in sewage sludge samples by liquid chromatography-tandem mass spectrometry: comparison of the efficiency of three extraction techniques.
This work presents a comparison of three extraction techniques -ultrasound-assisted extraction (USE), microwave-assisted extraction (MAE) and pressurized liquid extraction (PLE) - and evaluates their efficiency in the determination of quinolone antibiotics in sewage sludge samples. Extraction parameters for each technique were optimized using design of experiments, and the compounds were detected and quantified using liquid chromatography-tandem mass spectrometry (LC-MS/MS), operating in positive electrospray ionization (ESI) mode. The use of two selected reaction monitoring transitions for each compound allowed simultaneous quantification and identification in one run. Analytes were separated in less than 10 min. Marbofloxacin and cincophen were used as surrogates for amphoteric and acid quinolones, respectively. The limits of detection (LODs) were between 2 and 5 ng g(-1), and the limits of quantification (LOQs) were between 4 and 18 ng g(-1) for the various analytes. The inter- and intra-day variability was <7%. Due to the absence of certified reference materials (CRMs), the method was validated using matrix-matched calibration and a recovery assay with spiked samples. Recovery rates were between 97.9% and 104.8%. Statistical comparison demonstrated no significant differences between the three extraction techniques. The methods were successfully applied for the determination of quinolones in sewage sludge samples collected from different wastewater treatments plants (WWTPs) located in the province of Granada (Spain). The analytical methods developed here may be useful for the development of more in-depth studies on the occurrence and fate of these commonly used pharmaceuticals in WWTPs and in the environment.


[Yeasts associated with spontaneous fermentation processes in wines from Ribeiro. Analysis of homo/heterothallism and the killer system of S. cerevisiae strains].
A taxonomic analysis has been carried out concerning the different yeast species involved in spontaneous fermentations of Ribeiro wine. The results revealed the presence of 15 species belonging to 7 genera: Saccharomyces, Torulaspora, Dekkera, Saccharomycodes, Debaryomyces, Kloeckera and Candida. Within the genus Candida a high number of species was detected. The 28 isolated strains of S. cerevisiae showed with some exceptions, a remarkable sporulation ability and viability of the meiotic products. All the strains were homothallic. Among all S. cerevisiae strains, 6 exhibited killer K2 activity with higher isolation percentages in intermediate fermentations than in late fermentations.


Influence of endotoxin-induced acute lung injury on pulmonary innate and adaptive immunity.
The aim of our study was to analyze pulmonary innate and adaptive immune responses during endotoxemia-induced acute lung injury (ALI). Female BALB/c mice were challenged by endotoxin given intraperitoneally and followed for 24 h by unrestrained plethysmographic analysis. After this period, the mice were sacrificed by CO2 anesthesia and lung histopathology, pulmonary and serum levels of pro-inflammatory cytokines, numbers of lymphocyte subsets in blood and lung, and lung-derived macrophages (Mphi) were analyzed. Animals with endotoxemia demonstrated significant depression of tidal volumes indicating respiratory failure compared to control mice. Lung histopathology of endotoxin-exposed animals revealed alveolar leakage characterizing ALI. Pulmonary levels of interleukin (IL)-1beta, IL-6 and interferon (IFN)-gamma in animals with endotoxemia were significantly elevated, whereas serum levels of IL-6 only were increased. IFN-gamma was strongly expressed by lung-derived Mphi with high CD11b expression, and this subset significantly increased in the lungs after endotoxin challenge. Additionally, the numbers of lung-resident CD4+ and total T-lymphocytes were significantly reduced after challenge. These data suggest that endotoxemia-induced ALI is associated with exaggerated and sustained pulmonary innate immune responses partly mediated by activated Mphi, whereas adaptive immunity in the lungs is compromised.


Conversion of novel tannery sludge-derived biochar/TiO2 nanocomposite for efficient removal of Cr (VI) under UV light: photocatalytic performance and mechanism insight.
An investigation on the reduction of Cr (VI) pollutant from tannery effluents using TiO2, SB/TiO2, and c-SB/TiO2 nano photocatalysts was presented in this study. For the preparation of Biochar-based TiO2 photocatalyst (SB/TiO2), tannery sludge was utilized as a precursor. Hydrothermal pre-treatment was adopted to prepare chemically activated SB/TiO2 and SB/TiO2 nanocomposites. The morphology, crystal structure, optical properties, and elemental composition of the prepared catalysts were analyzed by XRD, FT-IR, SEM-EDX, BET analysis, ZPC, PL, TGA, and Raman spectroscopy. The band gap analysis of Photocatalyst was measured using a DRS instrument, and band gap energy of 3.39 eV was obtained for c-SB/TiO2 photocatalyst. The developed c-SB/TiO2 catalyst exhibits a larger specific surface area of 646.85 m2/g than TiO2 and SB/TiO2 (74.58 m2/g and 573.74 m2/g), respectively. The enhanced photocatalytic activity for the pollutant removal was achieved by the photocatalyst due to their wide band gap and effective charge separation. The kinetic rate constant was achieved in the pseudo-first-order model, which fits well for the reduction of Cr (VI). Furthermore, at the optimal conditions of 10 mg/L contaminant concentration, pH 2, and 0.5 g/L catalyst dosage, 98.56% reduction was observed after 180 min of reaction. The OH acts as a major removal pathway for Cr (VI) contaminants with more than 50% reduction in COD. This study proves that c-SB/TiO2 photocatalysts can remove toxic contaminants under UV light irradiation with good recycling performance up to 5 times.


Adjuvant immunochemotherapy with oral Tegafur/Uracil plus PSK in patients with stage II or III colorectal cancer: a randomised controlled study.
Intravenous fluorouracil and leucovorin is the standard adjuvant treatment for stage III colon cancer. However, oral adjuvant chemotherapy is attractive because it has low toxicity and greater convenience. We investigated the benefits of oral protein-bound polysaccharide K (PSK) with tegafur/uracil (UFT) as an adjuvant in stage II and III colorectal cancer. Patients were assigned to groups that received either 3 g PSK plus 300 mg UFT, or 300 mg UFT alone orally each day for a 2-year period following intravenous mitomycin C. Of 207 registered patients, 205 with stage II (n=123) or III (n=82) were analysed. The 5-year disease-free survival was 73.0% (95% CI 65.6-80.4%) with PSK (n=137) and 58.8% (95% CI 47.1-70.5%) in the controls (n=68) (P=0.016). Polysaccharide K reduced the recurrence by 43.6% (95% CI 4.5-66.7%) and mortality by 40.2% (95% CI -12.5 to 68.3%). The 5-year survival was 81.8% (95% CI 75.3-88.2%) in the PSK group and 72.1% (95% CI 61.4-82.7%) in the control group (P=0.056). In stage III patients, disease-free and overall survivals in patients receiving PSK were increased significantly: 60.0% (95% CI 47.1-72.9%) and 74.6% (95% CI 63.0-86.1%) in the PSK group as compared with 32.1% (95% CI 14.8-49.4%) and 46.4% (95% CI 28.0-64.9%) in the controls (P=0.002 and 0.003, respectively). Polysaccharide K prevented recurrence, particularly lung metastases (P=0.02; odds ratio 0.27; 95% CI 0.09-0.77). In the models, the presence of regional metastases (relative risk, 2.973; 95% CI 1.712-5.165; P<0.001), omission of PSK (relative risk, 2.106; 95% CI 1.221-3.633; P=0.007), and higher primary tumour (relative risk, 4.398; 95% CI 1.017-19.014; P=0.047) were each significant indicators of recurrence. Adverse effects were mild and compliance was good. Oral PSK with UFT reduced recurrence in stage II and III colorectal cancer, and increased survival in stage III.


Differential gene expression in experimental hepatocellular carcinoma induced by woodchuck hepatitis B virus.
Hepatitis B virus infection is closely linked to hepatocellular carcinoma (HCC), the pathological mechanism of hepatocarcinogenesis by this virus is not well understood. In order to gain further insight into the molecular mechanism of HCC, we constructed and screened a subtracted c-DNA library which was specific to HCC cells of a woodchuck infected with woodchuck hepatitis B virus. Among eight clones that were isolated based on their differential expressions, we determined nucleotide sequences of two genes whose expressions were most significantly stimulated in HCC. Our results indicate that these two genes appear to be woodchuck counterpart genes of hemopexin (HPX) and alpha-1 acid glycoprotein (AGP), suggesting that the expression of HPX and AGP genes are strongly augmented in tumor cells partly due to transcriptional regulation.


Absence of circulating natural killer (NK) cells in a child with erythrophagocytic lymphohistiocytosis lacking NK cell activity.
A 5-year-old girl who was diagnosed as having erythrophagocytic lymphohistiocytosis died at age 9 years. Peripheral lymphocytes from the patient persistently lacked natural killer (NK) cell activity during the 4-year observation period: the percent lysis values as measured by a 4-hr 51Cr release assay at a 40:1 effector:target ratio were below 1.0% against K562 and Molt-4 cells as compared with the normal lymphocyte value (mean +/- SD) of 46.2% +/- 5.8% and 43.9% +/- 6.7%, respectively. The patient's lymphocytes never developed NK cell activity by their incubation with target cells for longer time periods or by their stimulation with interferon-alpha, interleukin-2, or polyinosinic-polycytidilic acid. Single cell-in-agarose assay showed the absence of target-binding cells (TBCs): TBC numbers were below 0.3% as compared with the normal lymphocyte value of 8.1% +/- 1.3% (mean +/- SD). Flow cytometry showed a marked decrease in Leu-7+ cells (1.7%) and the absence of Leu-11+ cells (0.4%) in the peripheral blood. These results first demonstrate a case of erythrophagocytic lymphohistiocytosis in which there is the lack of NK cell activity due to the absence of circulating NK cells.


Association of renal insufficiency with treatments and outcomes in patients with acute coronary syndrome in China.
We aimed to analyze the association of estimated glomerular filtration rate (eGFR) levels of hospitalized patients with treatment decisions and clinical outcomes in Chinese patients with acute coronary syndrome (ACS). This was a secondary analysis study from CPACS-2 Program which was a trial of a quality improvement intervention in China and recruited 15,141 patients from 75 hospitals between October 2007 and August 2010. All patients were divided into three groups by the eGFR level on admission. The primary outcomes were several key performance indicators (KPIs) reflecting the management of ACS and the secondary outcomes were clinical outcomes. A total of 14,437 ACS patients were enrolled in this analysis. Among patients with reduced eGFR levels, fewer patients received appropriate medical therapy (p for trend <0.001) and fewer high-risk patients received coronary angiography (p for trend <0.001) compared to patients with a normal eGFR. Furthermore, 436 cases of death, 357 cases of cardiac death, 686 cases of major adverse cardiovascular events, and 198 cases of major bleeding episodes were reported. Patients with a worse eGFR level had significantly higher rates of death (p for trend <0.001), cardiac death (p for trend <0.001), major adverse cardiovascular events (p for trend <0.001) and major bleeding episodes (p for trend <0.001). Among Chinese ACS patients, those with renal insufficiency have a lower percentage of adherence to guideline-recommended treatments and worse clinical prognosis. Renal insufficiency is an important factor affecting guideline implementation in Chinese ACS patients. http://www.anzctr.org.au/default.aspx. Unique identifier: ACTRN12609000491268.


Interleukin-37 gene polymorphism and susceptibility to pulmonary tuberculosis among Iraqi patients.
Control of tuberculosis (TB) depends on a balance between host's immune factors and bacterial evasion strategies. Interleukin-37 (IL-37) is among the immunomodulatory factors that have been proposed to influence susceptibility to tuberculosis. A case-control study was conducted on 105 patients with pulmonary TB (37 active, 41 multi-drug resistant and 27 relapse) and 79 healthy controls to determine serum levels and single nucleotide polymorphisms (SNPs) of IL-37. The IL-37 level was assessed with an enzyme-linked immunosorbent kit, while DNA-sequencing was used to detect SNPs in the promoter region of IL37 gene. Median level of IL-37 was markedly increased in serum of TB patients compared to controls (325.0 vs. 169.1 pg/mL; p < 0.001). This increase was universally determined in subgroups of patients distributed according to gender, age groups, and clinical type of disease, while no significant differences were found between the subgroups in patients or controls. Analysis of receiver operating characteristic curve confirmed these findings and IL-37 occupied a very good area under the curve, which was 0.816 (95% CI = 0.744-0.888; p < 0.001). At a cut-off value of 185.6 pg/mL, the sensitivity and specificity of IL-37 were 81.0 and 82.3%, respectively. Of the nine detected SNPs (rs2466449 G/A, rs2466450 A/G, rs2723168 G/A, rs3811042 G/A, rs3811045 T/C, rs3811046 G/T, rs3811047 A/G, rs3811048 G/A and rs200782323 G/A), only rs3811048 showed a significant association with TB; the G allele showed a significantly decreased frequency in TB patients compared to controls (25.2 vs. 44.9%; OR = 0.41; p < 0.001). It was possible to assign five haplotypes, and three showed significant differences between patients and controls. Frequency of haplotype A-A-G-A-C-T-G-A-G (0.331 vs. 0.213; OR = 2.10; p = 0.015) was significantly increased in TB patients compared to controls. On the contrary, frequencies of haplotypes A-A-G-A-C-T-G-G-G (0.029 vs. 0.116; OR = 0.24; p = 0.01) and A-A-G-G-T-G-A-G-G (0.140 vs. 0.275; OR = 0.45; p = 0.015) were significantly decreased in patients. IL-37 was up-regulated in the serum of TB patients irrespective of their gender, age or clinical type of disease. SNPs in the promoter region of IL37 gene were proposed to be associated with susceptibility to TB.


A comparative study of collimation in bedside chest radiography for preterm infants in two teaching hospitals.
Unnecessary exposure of the abdomen, arms or head may lead to a substantial increase of the radiation dose in portable chest X-rays on the neonatal intensive care unit. The objective was to identify potential factors influencing inappropriate exposure of non-thoracic structures in two teaching hospitals. The study analysed 200 consecutive digital chest radiographs in 20 preterm neonates (mean gestation 25 ± 1 weeks). Demographical data, tube settings and exposure parameters were recorded. To grade the collimation, we used a scoring system with a maximum of 12 exposed non-thoracic structures. Length of gestation, age, the radiographer, years of experience in performing X-rays and the number of in situ catheters or lines, were correlated with collimation quality. There was no significant difference between the rates of optimal images obtained in the two hospitals (0.32 vs 0.39, n.s.). Scores showed that most suboptimal images had only mildly reduced image quality (1.40 ± 1.38 vs 1.20 ± 1.43, n.s.). Length of gestation or presence of surgical drains, catheters and tubes had no obvious effects on the exposure of non-thoracic structures. Large intra-individual variation in optimal collimation (14-86%) was noted for the radiographers in both hospitals; this was unrelated to their respective years of experience. In our study, the only identifiable factor influencing the collimation of portable chest radiographs in preterm infants was the radiographer's dedication and awareness. There were no apparent differences between the hospitals investigated. Exposure of non-thoracic structures was relatively frequent and mainly involved the proximal humeri.


Calcium uptake into guinea-pig trachealis: the effect of epithelium removal.
Removal of the epithelium from preparations of guinea-pig airways in vitro increases the responsiveness of the smooth muscle of normal and ovalbumin-sensitized animals to a number of contractile agents. To determine if epithelium removal results in an increase in Ca2+ entry into the smooth muscle, the effect of removing the epithelium on Ca2+ uptake into the trachealis smooth muscle was studied using a modified La3+-technique. KCl increased Ca2+ uptake in the presence and absence of the epithelium in control and sensitized animals. Methacholine did not promote Ca2+ uptake, whether or not the epithelium was present, in either control or sensitized animals. Ovalbumin did not stimulate Ca2+ uptake into the trachealis of sensitized animals. These results indicate that the increase in responsiveness of airway smooth muscle seen on epithelium removal is not a consequence of a facilitation of Ca2+ entry into the muscle. The increased responsiveness to methacholine in control animals, and to ovalbumin in preparations in tension studies in epithelium-free tissues from sensitized animals, cannot be explained by an increased availability of extracellular Ca2+ into the muscle, but, rather may reflect some other effect of the epithelium-derived modulatory factor.


Growth inhibition by danazol in a human endometrial cancer cell line with estrogen-independent progesterone receptors.
Since we recently found that danazol, an isoxazol derivative of ethinyltestosterone, has a growth-inhibitory effect on human endometrial cancer cells in primary culture, the effects of danazol on a human endometrial cancer cell line (IK-90 cells), which contains estrogen-independent progesterone receptors (PR), were investigated in the present study. The addition of danazol (1 nM-1 microM) in culture medium caused a decrease in the growth of IK-90 cells in a dose-dependent manner. Competitive binding studies showed that danazol effectively binds to PR in IK-90 cells, and the binding affinity for PR was estimated to be 6.0% of that of R5020. The addition of 1 microM danazol in culture medium resulted in a rapid and significant increase in nuclear PR with a concomitant decrease in cytoplasmic PR in the cells. These findings suggest that danazol has a growth-inhibitory effect on human endometrial adenocarcinoma cells directly through PR system in the cells.


Biological parameters on computed tomography guided fine needle aspiration biopsy from peripheral primary non-small cell lung cancer.
Computed tomography (CT) guided fine needle aspiration biopsy (CT-guided FNAB) represents the procedure of choice for diagnosing peripheral primary lung cancer before surgery. The aim of the present study was to assess the reliability of the immunocytochemical evaluation of biological parameters and DNA flow cytometry on cellular material obtained from non-small cell lung cancer (NSCLC) patients by CT-guided FNAB. Thirty consecutive CT-guided FNABs obtained from NSCLC patients were submitted both to the immunocytochemical evaluation of p53, Ki67, bcl-2 and to flow cytometric DNA analysis. p53, Ki67 and bcl-2 were assessable in 60% (18/30), 53% (16/30) and 48% (10/21) of the cases, respectively. Flow cytometric DNA analysis was performed in 19 out of the 30 cases and 74% (14/19) of the histograms were evaluable. Cytofluorimetric S-phase fraction (SPF), was obtained in 57% (8/14) of the cases. The results of the current study suggest that CT-guided FNAB from primary NSCLC patients may represent an effective practice for the evaluation of biologic parameters and could be useful as a preoperative procedure. The role of neoadjuvant chemotherapy in operable NSCLC is still under debate. We suppose that in the future the presurgical characterization of NSCLC could suggest the opportunity of a neoadjuvant systemic treatment aimed to improve the clinical outcome. Moreover, in locally advanced or metastatic NSCLC immunocytochemistry could help to predict the response to chemotherapy and/or radiotherapy, avoiding ineffective treatments and supporting the development of more rational therapies.


Antibiotic resistance of Helicobacter pylori in Australia and New Zealand: A systematic review and meta-analysis.
While the global prevalence of antibiotic-resistant Helicobacter pylori (H. pylori) is increasing, there is much regional variation, and local data are required to guide eradication therapy. We performed a systematic review and meta-analysis to determine rates of H. pylori antibiotic resistance in Australia and New Zealand. Random effects meta-analysis of data from 15 published studies and three published abstracts reporting prevalence of primary or secondary H. pylori antibiotic resistance in Australasia. PubMed, EMBASE, MEDLINE, PROSPERO, and the Cochrane Library were searched until August, 2020. Fifteen published studies and three published abstracts were identified; one study was excluded due to high risk of bias. Seventeen studies conducted between 1996 and 2013 were included in the final analysis, 12 reporting primary and five reporting secondary antibiotic resistance. Prevalence of primary resistance was clarithromycin 7.4% (95% confidence interval [CI], 5.3-9.7%), metronidazole 50.0% (95%CI, 23.9-56.1%), fluoroquinolones 3.7% (95%CI, 0.004-14.8%), and both amoxicillin and tetracycline <0.5%. Subgroup analysis (last 20 years) showed doubling of clarithromycin resistance to 16.1% (95%CI 11.2-21.7%) with other resistance stable. Prevalence of secondary resistance was high for all antibiotics, particularly clarithromycin 78.7% (95%CI, 64.1-90.1%) and metronidazole 68.3% (95%CI, 59.9-76.1%). The outcomes reveal an increase in primary H. pylori clarithromycin resistance since the year 2000, while metronidazole resistance has remained stable and primary resistance to amoxicillin, tetracycline, and fluoroquinolones is low. Rates of secondary resistance to metronidazole and clarithromycin are high. The results highlight the need for contemporary local data on antibiotic resistance in Australia and New Zealand.


Development of an evidence-based care bundle protocol for preventing falls in hospitalized children: Delphi study and trial test.
The aim of the study was to develop an evidence-based care bundle protocol for preventing falls in hospitalized children. Delphi method. Based on the four basic concepts of nursing, the theoretical dimension and item pool of the care bundle first draft were constructed by searching evidence and referring to the care bundle for preventing inpatient fall issues by the China Nursing Quality Promotion Alliance. The Delphi method and trial tests were used to select and adjust items of the care bundle to form the care bundle protocol for preventing falls in hospitalized children. The first draft care bundle for preventing falls in hospitalized children included 4 dimensions and 21 items. In the 2-round Delphi study, one item was removed, and three items were adjusted. One item was adjusted by trial test. Finally, the care bundle protocol for preventing falls in hospitalized children consisted of 4 dimensions and 20 items.


Trichothecene mycotoxins in kernels and head fusariosis susceptibility in winter triticale.
A single isolates ofFusarium graminearum Schwabe KF 366 andFusarium culmorum (W.G.Sm.) Sacc. KF 365 were used to infect 10 genotypes (9 lines and one cultivar) of winter triticale, 1 rye cultivar and 1 wheat cultivar, and amounts of mycotoxins in kernels were analysed at the same stage of development. One genotype of triticale CHD 353/79 and rye "Chodan" were found to be most resistant towards both species infection with lowest amount of mycotoxins (deoxynivalenol) content in kernels and also the lowest yield reduction. The most susceptible line CZR 142 cumulated in kernels about ten times higher amount of mycotoxins (up 53 mg DON/kg and 16 mg 3AcDON/kg, and 5 mg zearalenone/kg). GenerallyF, culmorum formed higher level of mycotoxins in kernels of infected heads thanF. graminearum. In kernels of more susceptible genotypes except deoxynivalenol, 3 acetyldeoxynivalenol and zearalenone also were present.


Rituximab as potential therapy for paraneoplastic cerebellar degeneration in pediatric Hodgkin disease.
Paraneoplastic cerebellar degeneration (PCD) is a rare neurological syndrome associated with lung cancer, breast adenocarcinoma,ovarian adenocarcinoma, and Hodgkin disease. It is rarely seen in pediatrics. We report a case of a 10-year-old boy with a 2-year prodrome that led to a diagnosis of PCD in association with stage IV Hodgkin disease. He received radiation and chemotherapy for his Hodgkin disease with resolution of his lymphoma. Based on promising data in adults on the efficacy of rituximab over other immuno suppressive agents in paraneoplastic disorders, he was treated with rituximab with marked improvement of the cerebellar syndrome.


Hollow Prussian blue with ultrafine silver nanoparticle agents (Ag-HPB) integrated sensitive and flexible biosensing platform with highly enzyme loading capability.
Herein, the hollow Prussian blue with ultra-small silver nanoparticle agents (Ag-HPB) was prepared by the coating-etching method by applying Prussian blue (PB) coating on Ag nanoparticles (Ag NPs) and diffusing Ag NPs into the PB framework. The flexible biosensing platform based on Ag-HPB nanocomposites incorporated the excellent electrical conductivity of Ag NPs and the superior enzyme loading capacity of the hollow structure, which significantly enhanced its sensing performance. Subsequently, take glucose oxidase (GOx) and acetylcholinesterase (AChE) as examples. The sensing platform displayed a good sensitive response to glucose (Glu) (24.37 μA mM-1 cm-2) and a considerable limit of detection (LOD) for trichlorfon (TCF) as 2.28 pg/mL while exhibiting high stability and good reproducibility. Moreover, it can be applied to monitor trichlorfon in apple samples. Promisingly, the Ag-HPB prepared by the coating-etching strategy provides a reliable strategy for further development of sensitive and flexible biosensing platforms with excellent electrical conductivity and high enzyme loading.


Cleaning of oval canals using ultrasonic or sonic instrumentation.
This study investigated the effect of direction of file oscillation on the amount of debris and smear remaining within oval canals. Sixty-five lower premolars were allocated to six experimental groups (n = 10) and one control group (n = 5). Canals were prepared using Gates Glidden burs and hand instruments. Preparations were completed using either an ultrasonic size 15 K file or a sonic size 15 shaper file. Three preparation techniques were used: (a) file oscillation toward oval recesses within the canal, (b) file oscillation at right angles to the recesses, and (c) rotational movement of handpiece. Canals were examined under the scanning electron microscope and blindly scored for debris and smear layer. Log linear analysis revealed no significant difference (p > 0.05) between sonic or ultrasonic instrumentation as to debris or smear layer remaining. File oscillation directed toward oval recesses left the least amount of debris (p < 0.05). Smear layer remained unaffected by all techniques.


Synthesis of UDP-glucose derivatives modified at the 3-OH as potential chain terminators of beta-glucan biosynthesis.
A series of UDP-D-glucose derivatives and precursors that have been modified at C-3 were synthesised from D-glucose as potential chain terminators of beta-glucan biosynthesis. None of the UDP-derivatives or the precursors tested displayed significant anti-fungal activity in a series of germination assays on the dermatophyte Trichophyton rubrum.


Streptococcus mutans isolated from children with severe-early childhood caries form higher levels of persisters.
Dental caries is the most common chronic infectious disease in children. Streptococcus mutans, the main cariogenic bacterial species, produces persisters, nongrowing dormant variants of regular cells associated with chronicity of diseases. We hypothesized that the recurrent nature of caries, particularly within populations with high-caries risk, is due partly to specific phenotypic features of S. mutans such as its ability to form persisters. We aimed to investigate the genotypic and phenotypic differences between the S. mutans from children with severe early-childhood caries (S-ECC) and those without caries. S. mutans from plaque samples of caries-free (CF) and S-ECC children were tested for their ability to adapt to a lethal pH in an acid tolerance response assay. The persister levels of S. mutans isolates was quantified in both groups. S. mutanswas identified in all 23 S-ECC but only 6 of the 21 CF subjects. In most subjects, only one dominant S. mutans genotype was detected. No statistically significant differences in the mean survival percentage of S. mutans were observed between the two groups at a lethal pH of 3.5. However, the dominant genotype within a particular S-ECC subject exhibited a higher percentage of cell survival compared to those in the CF group. In S-ECC patients, S. mutans isolates displayed a ∼15-fold higher persistence phenotype than S. mutans isolates from CF patients. The ability of S. mutans to produce high levels of persisters may contribute to part of an individual's ability to control caries disease activity and recurrent lesions.


In MALDI-Mass Spectrometry Imaging on Formalin-Fixed Paraffin-Embedded Tissue Specimen Section Thickness Significantly Influences m/z Peak Intensity.
In matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) standardized sample preparation is important to obtain reliable results. Herein, the impact of section thickness in formalin-fixed paraffin embedded (FFPE) tissue microarrays (TMA) on spectral intensities is investigated. TMAs consisting of ten different tissues represented by duplicates of ten patients (n = 200 cores) are cut at 1, 3, and 5 μm. MSI analysis is performed and mean intensities of all evaluable cores are extracted. Measurements are merged and mean m/z intensities are compared. Visual inspection of spectral intensities between 1, 3, and 5 μm reveals generally higher intensities in thinner tissue sections. Specifically, higher intensities are observed in the vast majority of peaks (98.6%, p < 0.01) in 1 μm compared with 5 μm sections. Note that 28.4% and 2.1% of m/z values exhibit a at least two- and threefold intensity difference (p < 0.01) in 1 μm compared to 5 μm sections, respectively. A section thickness of 1 μm results in higher spectral intensities compared with 5 μm. The results highlight the importance of standardized protocols in light of recent efforts to identify clinically relevant biomarkers using MSI. The use of TMAs for comparative analysis seems advantageous, as section thickness displays less variability.


Reflex sympathetic dystrophy: an underreported syndrome in children?
Two cases of reflex sympathetic dystrophy are described in a 13-year-old and a 10-year-old girl. In the latter, symptoms occurred consecutively in the right leg, the left foot and the left hand. In contrast to the usual findings in adults, bone scintigraphy showed decreased radio-isotope uptake in the affected area during the early phase. An electromyography performed in the younger patient showed slower nerve conduction velocity in the affected limb. The younger girl improved following physical therapy and calcitonin injections, while the older patient favorably responded to sympathetic blockade.


A Proposal for Separation of Nuclear Atypia and Architectural Atypia in Bethesda Category III (AUS/FLUS) Based on Differing Rates of Thyroid Malignancy.
Bethesda category III (atypia of undetermined significance/follicular lesion of undetermined significance) includes sparsely cellular specimens with nuclear atypia (3N) and/or architectural atypia (3A). This study investigates whether the two types of atypia have different rates of malignancy (ROMs). Cytologic and histologic diagnoses of resected thyroid nodules were recorded. ROM was calculated for all Bethesda categories and for 3N and 3A subcategories. Possible noninvasive follicular thyroid neoplasms with papillary-like nuclear features were reviewed and removed from malignancies, and ROM was recalculated. A total of 1,396 nodules were included. ROM of 3N (33.3%-26.0%) was higher than 3A (7.7%-5.0%) (P < .0001) and was similar to suspicious for follicular neoplasm (25.0%-20.3%) (P = .3). ROM of 3A approached benign (2.4%-1.5%) (P = .02). Strong consideration should be given to separating 3N (nuclear atypia with higher risk for papillary thyroid carcinoma) from 3A (architectural atypia with higher chance of being benign) to convey different ROMs.


Hyperactivity of fibroblasts cultured from psoriatic skin: I. Faster proliferation and effect of serum withdrawal.
Fibroblasts were cultured from the involved and uninvolved forearm skin of patients with severe generalized psoriasis and compared with those from the forearms of normal controls of similar ages. Thirteen strains were obtained from involved skin (PSA strains) and sixteen strains from uninvolved skin (PSB), with thirteen control strains (NSF). Outgrowth of fibroblasts from the psoriatic skin explants was slightly quicker than from control skin and the average proliferation rates of passaged strains were PSA 144, PSB 134 and NSF 94 (P less than 0.05). Psoriatic fibroblasts were abnormally dependent on serum for anchorage. In serum-free medium many cells rounded up and were only loosely attached to the substratum. This effect was rapid, reversible and not corrected by adding fibronectin. Cell attachment assays showed only small differences between the psoriatic and normal fibroblasts and the main effect of serum withdrawal appeared to be on spreading rather than attachment. These data suggest that the dermis of both involved and uninvolved psoriatic skin is abnormal and that the hyperactivity persists in vitro. Our findings seem most compatible with a hyperproliferative reaction of both epidermis and dermis to an extracutaneous, perhaps vascular, stimulus.


17β-Estradiol attenuates secondary injury through activation of Akt signaling via estrogen receptor alpha in rat brain following subarachnoid hemorrhage.
Apoptosis is implicated in vasospasm and the long-term sequelae of subarachnoid hemorrhage (SAH). This study tested the hypothesis that attenuation of SAH-induced apoptosis after 17β-estradiol (E2) treatment is associated with an increase in phosphorylation of Akt via estrogen receptor-α (ER-α) in rats. We examined the expression of phospho-Akt, ERα and ERβ, and apoptosis in cerebral cortex, hippocampus, and dentate gyrus in a two-hemorrhage SAH model in rats. We subcutaneously implanted other rats with a silicone rubber tube containing E2; they received daily injections of nonselective estrogen receptor antagonist (ICI 182,780), selective ERα-selective antagonist (methyl-piperidino-pyrazole), or ERβ-selective antagonist (R,R-tetrahydrochrysene) after the first hemorrhage. At 7 d after the first SAH, protein levels of phospho-Akt and ERα were significantly decreased and caspase-3 was significantly increased in the dentate gyrus. The cell death assay revealed that DNA fragmentation was significantly increased in the dentate gyrus. Those actions were reversed by E2 and blocked by ICI 182,780 and methyl-piperidino-pyrazole, but not R,R-tetrahydrochrysene. However, there were no significant changes in the expression of the protein levels of phospho-Akt, ERα, ERβ, and caspase-3, and DNA fragmentation after SAH. The present study shows that a beneficial effect of E2 in attenuating SAH-induced apoptosis is associated with activation of the expression of phospho-Akt and ERα, and alteration in caspase-3 protein expression via an ERα-dependent mechanism in the dentate gyrus. These data support further the investigation of E2 in the treatment of SAH in humans.


Modifiable risk factors for glioblastoma: a systematic review and meta-analysis.
Glioblastoma (GBM) is the most common and aggressive glioma histological subtype, associated with high disability and poor survival. The etiology of this condition is still mostly unknown, and evidence about risk factors is elusive. The aim of this study is to identify modifiable risk factors for GBM. Electronic search was performed by two reviewers independently using the keywords and MeSH terms 'glioblastoma' OR 'glioma' OR 'brain tumor' AND 'risk factor'. The inclusion criteria were (1) observational studies or experimental studies on humans, (2) studies assessing the association between glioblastoma and exposure to modifiable conditions, and (3) studies published in English or Portuguese. Studies on the pediatric population or about exposure to ionizing radiation were excluded. A total of 12 studies were included. Seven were case-control studies, and five were cohort studies. The risk factors assessed included body mass index, alcohol consumption, exposure to magnetic fields, diabetes mellitus type 2 (DM2), and use of non-steroidal anti-inflammatory drugs (NSAID). No significant link was found between GBM incidence and DM2 or magnetic field exposure. On the other hand, higher BMI, alcohol consumption, and NSAID use demonstrated a protective effect on GMB risk. However, given the limited number of studies, it is not possible to obtain a behavioral recommendation; instead, these findings are relevant to guide future basic scientific studies on GBM oncogenesis.


Application of Balloon-Expandable Transcatheter Heart Valve in Bicuspid Aortic Valve.
Bicuspid aortic valve (BAV) remains challenging in transcatheter aortic valve replacement (TAVR) because of unfavorable anatomy. New-generation balloon-expandable valve (BEV) appears to be a valid alternative to surgery, especially in some Asian countries with a higher prevalence of BAV. This tutorial review summarizes current thinking about how to plan and implant BEV in BAVs using versatile techniques. First, the authors depict the main morphological characteristics of BAVs and their effects on the TAVR procedure. Next, the authors provide preprocedural analysis on sizing, obtaining the optimal deployment projection, and how to simplify valve-crossing. Finally, the authors provide step-by-step guidance on how to deploy the BEVs with evolved iterations in terms of specific anatomies, calcified annulus, and giant annulus.


Labeling of human erythrocyte membranes with eosin probes used for protein diffusion measurements: inhibition of anion transport and photo-oxidative inactivation of acetylcholinesterase.
The binding of eosin-isothiocyanate (eosin-NCS) and iodoacetamido-eosin (IA-eosin) to band 3 proteins in the membrane of human erythrocytes is characterized by studying the effect of these probes on the anion transport system. Although the unbrominated fluorescein precursors do not affect anion transport, both eosin labels are strong inhibitors of sulphate exchange in intact erythrocytes. 50% inhibition is obtained by binding 4.7 . 10(5) or 6.0 . 10(5) molecules/cell for eosin-NCS and IA-eosin, respectively. Both eosin probes are irreversibly bound and occupy common binding sites with 4,4-diisothiocyano-1,2-diphenyl-ethane-2,2'disulfonic acid (H2DIDS), although other sites are labeled as well. The inhibition of anion transport is light independent and can therefore not be attributed to a photosensitizing action of the eosin probes. Both eosin derivatives, however, inactivate acetylcholinesterase upon illumination of air-equilibrated samples of hemoglobin-free labeled ghosts. The inactivation of the enzyme is accompanied by the formation of protein aggregates as visualized by polyacrylamide gel electrophoresis. These effects are not observed when intact erythrocytes are illuminated in the presence of eosin probes suggesting a protective effect of hemoglobin during the labeling procedure. Protection of ghosts from photo-oxidation is achieved by displacing air with argon. These results are discussed in relation to the use of these and similar probes to measure protein diffusion in membranes.