Trapping toxins within lipid droplets is a resistance mechanism in fungi.
Lipid droplets (LDs) act as intracellular storage organelles in most types of cells and are principally involved in energy homeostasis and lipid metabolism. However, the role of LDs in resistance to toxins in fungi remains largely unknown. Here, we show that the trapping of endogenous toxins by LDs is a self-resistance mechanism in the toxin producer, while absorbing external lipophilic toxins is a resistance mechanism in the toxin recipient that acts to quench the production of reactive oxygen species. We found that an endolichenic fungus that generates phototoxic perylenequinones (PQs) trapped the PQs inside LDs. Using a model that incorporates the fungicidal action of hypocrellin A (HA), a PQ derivative, we showed that yeast cells escaped killing by trapping toxins inside LDs. Furthermore, LD-deficient mutants were hypersusceptible to HA-mediated phototoxins and other fungicides. Our study identified a previously unrecognised function of LDs in fungi that has implications for our understanding of environmental adaptation strategies for fungi and antifungal drug discovery.


Familial central precocious puberty: two novel MKRN3 mutations.
Paternally inherited loss-of-function mutations in MKRN3 underlie central precocious puberty (CPP). We describe clinical and genetic features of CPP patients with paternally inherited MKRN3 mutations in two independent families. The single coding exon of MKRN3 was analyzed in three patients with CPP and their family members, followed by segregation analyses. Additionally, we report the patients' responses to GnRH analog treatment. A paternally inherited novel heterozygous c.939C>G, p.(Ile313Met) missense mutation affecting the RING finger domain of MKRN3 was found in a Finnish girl with CPP (age at presentation 6 years). Two Polish siblings (a girl presenting with B2 at the age of 4 years and a boy with adult size testes at the age of 9 years) had inherited a novel heterozygous MKRN3 mutation c.1237_1252delGGAGACACATGCTTTT p.(Gly413Thrfs*63) from their father. The girls were treated with GnRH analogs, which exhibited suppression of the hypothalamic-pituitary-gonadal axis. In contrast, the male patient was not treated, yet he reached his target height. We describe two novel MKRN3 mutations in three CPP patients. The first long-term data on a boy with CPP due to an MKRN3 mutation questions the role of GnRH analog treatment in augmenting adult height in males with this condition. We describe the genetic cause for central precocious puberty (CPP) in two families. This report adds two novel MKRN3 mutations to the existing literature. One of the mutations, p.(Ile313Met) affects the RING finger domain of MKRN3, which has been shown to be important for repressing the promoter activity of KISS1 and TAC3. We describe the first long-term observation of a male patient with CPP due to a paternally inherited MKRN3 loss-of-function mutation. Without GnRH analog treatment, he achieved an adult height that was in accordance with his mid-parental target height.


Hippocampal Transections for Epilepsy.
Multiple hippocampal transection (MHT) is a novel surgical procedure that serves to disrupt seizure propagation fibers within the hippocampus without impairing verbal memory or the loss of stem cells. Given the paucity of literature regarding the utility and long-term outcome of MHT, a review is presented of the current literature to support the utility of this procedure in the treatment of intractable temporal lobe epilepsy. Long-term outcome analysis of this technique has been reported by 2 independent groups. Both groups used intraoperative electrocorticography. All patients underwent multiple subpial transection on the neocortex and MHT on the hippocampus.


[Arthritis of the Medial Knee Joint Compartment].
23 % of all persons older than 65 years suffer from osteoarthritis of the medial compartment of the knee joint, a very common situation in orthopaedic practice 1. As a result of the demographic trend the number of patients is expected to increase in the future. Based on specific joint biomechanics and kinematics the medial knee joint compartment is more frequently affected than the lateral. Only an understanding of the functional anatomy and underlying pathology allows a critical evaluation of different available conservative and operative treatment options. This article gives an overview of diagnostic and therapeutic strategies of osteoarthritis of the medial knee joint. Frequently performed surgeries, e.g. high tibial osteotomy (HTO), unicompartmental knee arthroplasty (UKA) and total knee arthroplasty (TKA) will be presented in a comparative manner. The actual scientific evidence will be given with the goal of an evidence based therapy that is adopted to stage and pathology of osteoarthritis of the medial compartment of the knee joint.


Fasting-mimicking diet blocks triple-negative breast cancer and cancer stem cell escape.
Metastatic tumors remain lethal due to primary/acquired resistance to therapy or cancer stem cell (CSC)-mediated repopulation. We show that a fasting-mimicking diet (FMD) activates starvation escape pathways in triple-negative breast cancer (TNBC) cells, which can be identified and targeted by drugs. In CSCs, FMD lowers glucose-dependent protein kinase A signaling and stemness markers to reduce cell number and increase mouse survival. Accordingly, metastatic TNBC patients with lower glycemia survive longer than those with higher baseline glycemia. By contrast, in differentiated cancer cells, FMD activates PI3K-AKT, mTOR, and CDK4/6 as survival/growth pathways, which can be targeted by drugs to promote tumor regression. FMD cycles also prevent hyperglycemia and other toxicities caused by these drugs. These data indicate that FMD has wide and differential effects on normal, cancer, and CSCs, allowing the rapid identification and targeting of starvation escape pathways and providing a method potentially applicable to many malignancies.


Studies on the interaction between ethanol and amfonelic acid.
Amfonelic acid (AFA), a non-amphetamine central stimulant dose-dependently reduced the hypnotic effect of ethanol in C57B1/6 mice. It did not enhance the elimination of ethanol. Amfonelic acid failed to modify the duration of pentobarbitone-induced hypnosis or the ethanol-induced hypothermia in these animals. Combined treatment with amfonelic acid and a lipophilic alpha 1-adrenoceptor agonist was not more effective than amfonelic acid alone in blocking ethanol hypnosis. The stimulation of locomotor activity by amfonelic acid in C57B1/6 mice was more sensitive to the blocking effect of ethanol than stimulation induced by d-amphetamine. The blocking effect of amfonelic acid, but not that of d-amphetamine, on the effects of ethanol developed tolerance. In pimozide-pretreated mice, amfonelic acid failed to reduce the ethanol-induced hypnosis. Hence it appears that dopamine (DA) released by amfonelic acid is responsible for its antagonism of ethanol. However, though amfonelic acid acted as a strong releaser of DA in Swiss-Webster, CD-1, DBA-2 and BALB/c mice, in these strains it failed to reduce the effect of ethanol. Moreover, methylphenidate, a dopaminergic stimulant, which acts by a mechanism similar to that of amfonelic acid was not effective in reducing the hypnotic effect of ethanol in C57B1/6 mice. For these reasons, additional mechanisms may have to be considered to explain this strain-dependent effect of amfonelic acid.


LGN-CNN: A biologically inspired CNN architecture.
In this paper we introduce a biologically inspired Convolutional Neural Network (CNN) architecture called LGN-CNN that has a first convolutional layer composed of a single filter that mimics the role of the Lateral Geniculate Nucleus (LGN). The first layer of the neural network shows a rotational symmetric pattern justified by the structure of the net itself that turns up to be an approximation of a Laplacian of Gaussian (LoG). The latter function is in turn a good approximation of the receptive field profiles (RFPs) of the cells in the LGN. The analogy with the visual system is established, emerging directly from the architecture of the neural network. A proof of rotation invariance of the first layer is given on a fixed LGN-CNN architecture and the computational results are shown. Thus, contrast invariance capability of the LGN-CNN is investigated and a comparison between the Retinex effects of the first layer of LGN-CNN and the Retinex effects of a LoG is provided on different images. A statistical study is done on the filters of the second convolutional layer with respect to biological data. In conclusion, the model we have introduced approximates well the RFPs of both LGN and V1 attaining similar behavior as regards long range connections of LGN cells that show Retinex effects.


Restoring gluconeogenesis by TEF inhibited proliferation and promoted apoptosis and immune surveillance in kidney renal clear cell carcinoma.
Kidney renal clear cell carcinoma (KIRC) is the major histological subtype of kidney tumor which covers approximately 80% of the cases. Although various therapies have been developed, the clinical outcome remains unsatisfactory. Metabolic dysregulation is a key feature of KIRC, which impacts progression and prognosis of the disease. Therefore, understanding of the metabolic changes in KIRC is of great significance in improving the treatment outcomes. The glycolysis/gluconeogenesis genes were analyzed in the KIRC transcriptome from the Cancer Genome Atlas (TCGA) by the different expression genes (DEGs) test and survival analysis. The gluconeogenesis-related miRNAs were identified by ImmuLncRNA. The expression levels of indicated genes and miRNAs were validated in KIRC tumor and adjunct tissues by QPCR. The effects of miR-4477b and PCK1 on cell proliferation and apoptosis were examined using the cell viability assay, cell apoptosis assay, and clone information. The interaction of miR-4477b with TEF was tested by the luciferase report gene assay. The different gluconeogenesis statuses of tumor cells and related signatures were investigated by single-cell RNA sequencing (scRNA-seq) analysis. The 11 gluconeogenesis genes were found to be suppressed in KIRC (referring as PGNGs), and the less suppression of PGNGs indicated better survival outcomes. Among the 11 PGNGs, we validated four rate-limiting enzyme genes in clinical tumor patients. Moreover, restoring gluconeogenesis by overexpressing PCK1 or TEF through miR-4477b inhibition significantly inhibited tumor cell proliferation, colony formation, and induced cell apoptosis in vitro. Independent single-cell RNA sequencing (scRNA-seq) data analysis revealed that the tumor cells had high levels of PGNG expression (PGNG + tumor cells) represented a phenotype of early stage of neoplasia and prompted immune surveillance. Our study suggests that the deficiency of gluconeogenesis is a key metabolic feature of KIRC, and restoring gluconeogenesis could effectively inhibit the proliferation and progression of KIRC cells.


Effect of eicosapentaenoic acid on triacylglycerol transport in CaCo-2 cells.
The human intestinal cell line, CaCo-2, was used to study the effect of the n-3 fatty acid, eicosapentaenoic acid, on triacylglycerol secretion. In cells incubated with 250 microM eicosapentaenoic acid, the incorporation of [3H]glycerol into triacylglycerols secreted into the medium was decreased by 58% compared to cells incubated with 250 microM oleic acid. The incorporation of [3H]glycerol into cellular triacylglycerols was decreased 32% in cells incubated with eicosapentaenoic acid. In cells preincubated with [3H]glycerol to label existing triacylglycerols, the rates of secretion of preformed triacylglycerols were similar in response to the addition of either fatty acid. Initial uptake rates of the n-3 fatty acid were higher than for oleic acid. Both eicosapentaenoic acid and oleic acid were minimally oxidized to CO2. Oleic acid was predominantly incorporated into cellular triacylglycerols (62% vs. 47%), whereas more eicosapentaenoic acid was incorporated into cellular phospholipids (46% vs. 30%). Phospholipids of microsomes prepared from cells incubated with eicosapentaenoic acid were enriched in this fatty acid. The rate of synthesis of triacylglycerol and diacylglycerol acyltransferase activities were significantly less in microsomes prepared from cells incubated with eicosapentaenoic acid. Triacylglycerol mass secreted by CaCo-2 cells incubated with either fatty acid was similar. In CaCo-2 cells, eicosapentaenoic acid decreases the synthesis and secretion of newly synthesized triacylglycerol without decreasing the secretion of triacylglycerol mass. Modification of microsomal membrane phospholipid fatty acid composition is associated with a decrease in microsomal triacylglycerol synthesis and diacylglycerol acyltransferase activities.


Epstein-Barr virus in somatic cell hybrids between mouse cells and human nasopharyngeal carcinoma cells.
Somatic cell hybrids between mouse cells and cells derived directly from NPC biopsies were produced in order to study the association of the Epstein-Barr virus (EBV) genome and the expression of Epstein-Barr nuclear antigen (EBNA) with the human chromosome(s). All attempts to correlate the presence of EBV-DNA and the expression of EBNA with the presence of a particular human chromosome(s) showed that the segregation of EBV-DNA or of EBNA and human chromosomes was dysconcordant. The data, therefore, suggest that in the hybrids studied the presence of EBA-DNA is not determined by the presence of a specific human chromosome.


Quantification of SNAP-25 with mass spectrometry and Simoa: a method comparison in Alzheimer's disease.
Synaptic dysfunction and degeneration are central to Alzheimer's disease (AD) and have been found to correlate strongly with cognitive decline. Thus, studying cerebrospinal fluid (CSF) biomarkers reflecting synaptic degeneration, such as the presynaptic protein synaptosomal-associated protein 25 (SNAP-25), is of importance to better understand the AD pathophysiology. We compared a newly developed Single molecule array (Simoa) immunoassay for SNAP-25 with an in-house immunoprecipitation mass spectrometry (IP-MS) method in a well-characterized clinical cohort (n = 70) consisting of cognitively unimpaired (CU) and cognitively impaired (CI) individuals with and without Aβ pathology (Aβ+ and Aβ-). A strong correlation (Spearman's rank correlation coefficient (rs) > 0.88; p < 0.0001) was found between the Simoa and IP-MS methods, and no statistically significant difference was found for their clinical performance to identify AD pathophysiology in the form of Aβ pathology. Increased CSF SNAP-25 levels in CI Aβ+ compared with CU Aβ- (Simoa, p ≤ 0.01; IP-MS, p ≤ 0.05) and CI Aβ- (Simoa, p ≤ 0.01; IP-MS, p ≤ 0.05) were observed. In independent blood samples (n = 32), the Simoa SNAP-25 assay was found to lack analytical sensitivity for quantification of SNAP-25 in plasma. These results indicate that the Simoa SNAP-25 method can be used interchangeably with the IP-MS method for the quantification of SNAP-25 in CSF. Additionally, these results confirm that CSF SNAP-25 is increased in relation to amyloid pathology in the AD continuum.


Cost-effective analysis of transcatheter aortic valve replacement in patients with severe symptomatic aortic stenosis: A prospective multicenter study.
Transcatheter aortic valve replacement (TAVR) for severe symptomatic aortic stenosis (AS) does not benefit all patients. We performed a prospective multicenter study to investigate the cost-effectiveness of TAVR in a Japanese cohort. We prospectively enrolled 110 symptomatic patients with severe AS who underwent TAVR from five institutions. The quality of life measurement (QOL) was performed for each patient before and at 6 months after TAVR. Patients without an improvement in QOL at 6 months after TAVR were defined as non-responders. Pre-TAVR higher QOL, higher clinical frailty scale predicted the non-responders. Three models, 1) conservative treatment for all patients strategy, 2) TAVR for all patients strategy, and 3) TAVR for a selected patient strategy who is expected to be a responder, were simulated. Lifetime cost-effectiveness was estimated using incremental cost-effectiveness ratio (ICER) and cost per quality-adjusted life-year (QALY) gained. In comparison to conservative therapy for all patients, ICER was estimated to be 5,765,800 yen/QALY for TAVR for all patients and 2,342,175 yen/QALY for TAVR for selected patient strategy patients, which is less than the commonly accepted ICER threshold of 5,000,000 yen/QALY. TAVR for selected patient strategy model is more cost-effective than TAVR for all patient strategy without reducing QOL in the Japanese healthcare system. TAVR for selected patient strategy has potential benefit for optimizing the TAVR treatment in patients with high frailty and may direct our resources toward beneficial interventions.


The effect of bilateral kainic acid-induced lateral habenula lesions on dopamine-mediated behaviors.
Bilateral kainic acid-induced lateral habenula lesions produced dose specific alterations in dopamine mediated behaviors in the rat. Intermediate doses of apomorphine and amphetamine produced potentiated stereotypic responsiveness while higher doses produced a behavioral response similar to that observed in sham-operated and virgin controls. Autoreceptor specific doses of apomorphine and its associated behavioral hypoactivity was unaffected by lesions. Animals with lesions did not exhibit the response potentiation normally induced by chronic haloperidol treatment.


Cerebral mitochondrial redox states during metabolic stress in the immature rat.
The brain mitochondrial NAD+/NADH ratio, as a reflection of the oxidation-reduction (redox) state of cellular compartment, was determined under conditions of hypoxia, anoxia, hypoxia-ischemia, complete ischemia and hypoglycemia in immature rats. NAD+/NADH ratios were calculated from changes in the concentrations of specific oxidative substrates and calculated intracellular pH during cerebral metabolic stress. The results suggest that the use of the acetoacetate/beta-hydroxybutyrate substrate couple provides a more accurate prediction of the mitochondrial redox state under adverse conditions than use of the alpha-ketoglutarate/glutamate couple. It is possible that the mitochondrial oxidation seen with the latter substrate couple during cerebral metabolic stress might reflect a population of cells (neurons or glia) which are substrate-deprived relative to the rest of the brain in the setting of metabolic stress produced by oxygen deficiency.


Distinct immune responses of recombinant plasmid DNA replicon vaccines expressing two types of antigens with or without signal sequences.
Here, DNA replicon vaccines encoding the Hc domain of botulinum neurotoxin serotype A (AHc) or the receptor binding domain of anthrax protective antigen (PA4) with or without signal sequences were evaluated in mice. Strong antibody and protective responses were elicited only from AHc DNA vaccines with an Ig κ signal sequence or tissue plasminogen activator signal sequence. Meanwhile, there were no differences in total antibody responses or isotypes, lymphocyte proliferative responses, cytokine profiles and protective immune responses with the PA4 DNA vaccines with or without a signal sequence. Therefore, use of targeting sequences in designing DNA replicon vaccines depends on the specific antigen.


Ordinary varieties with trivial canonical bundle are not uniruled.
We prove that smooth, projective, K-trivial, weakly ordinary varieties over a perfect field of characteristic p > 0 are not geometrically uniruled. We also show a singular version of our theorem, which is sharp in multiple aspects. Our work, together with Langer's results, implies that varieties of the above type have strongly semistable tangent bundles with respect to every polarization.


Glycogen synthase kinase-3beta inhibition preserves hematopoietic stem cell activity and inhibits leukemic cell growth.
Ex vivo expansion of cord blood cells generally results in reduced stem cell activity in vivo. Glycogen synthase kinase-3beta (GSK-3beta) regulates the degradation of beta-catenin, a critical regulator of hematopoietic stem cells (HSCs). Here we show that GSK-3beta inhibition activates beta-catenin in cord blood CD34(+) cells and upregulates beta-catenin transcriptional targets c-myc and HoxB4, both known to regulate HSC self-renewal. GSK-3beta inhibition resulted in delayed ex vivo expansion of CD34(+) cells, yet enhanced the preservation of stem cell activity as tested in long-term culture with bone marrow stroma. Delayed cell cycling, reduced apoptosis, and increased adherence of hematopoietic progenitor cells to bone marrow stroma were observed in these long-term cultures treated with GSK-3beta inhibitor. This improved adherence to stroma was mediated via upregulation of CXCR4. In addition, GSK-3beta inhibition preserved severe combined immunodeficiency (SCID) repopulating cells as tested in the nonobese diabetic/SCID mouse model. Our data suggest the involvement of GSK-3beta inhibition in the preservation of HSC and their interaction with the bone marrow environment. Methods for the inhibition of GSK-3beta may be developed for clinical ex vivo expansion of HSC for transplantation. In addition, GSK-3beta inhibition suppressed leukemic cell growth via the induction of apoptosis mediated by the downregulation of survivin. Modulators of GSK-3beta may increase the range of novel drugs that specifically kill leukemic cells while sparing normal stem cells.


Brain glucose levels in portacaval-shunted rats with chronic, moderate hyperammonemia: implications for determination of local cerebral glucose utilization.
Rates of glucose utilization (lCMRglc) in many structures of the brain of fed, portacaval-shunted rats, when assayed with the [14C]deoxyglucose (DG) method in our laboratory, were previously found to be unchanged (30 of 36 structures) or depressed (6 structures) during the first 4 weeks after shunting, but to rise progressively to higher than normal values in 25 of 36 structures from 4-12 weeks. In contrast, lCMRglc, when assayed with the [14C]glucose method in another laboratory, was depressed in most structures of brains of 4-8-week shunted rats that had relatively high brain ammonia levels. There was a possibility that the increases in lCMRglc obtained with the [14C]DG method may have been artifactual, due, in part, to a change in brain glucose content which could alter the value of the lumped constant of the DG method. Brain glucose levels of shunted rats were, therefore, assayed by both direct chemical measurement in freeze-blown samples and by determination of steady-state brain:plasma distribution ratios for [14C]methylglucose; the methylglucose distribution ratio varies as a function of plasma and tissue glucose contents. Within a week after shunting, ammonia levels in blood and brain rose to 0.25-0.30 mM and 0.35-0.70 mumol/g, respectively, and mean plasma glucose levels fell from 9-10 mM to 7.4-8.5 mM, and then remained nearly constant. Brains of fed-shunted rats had normal glycogen levels and stable but moderately reduced glucose contents between 1 and 12 weeks (i.e., 1.9-2.2 mumol/g). [14C]Methylglucose distribution ratios were essentially the same as those in controls in 22 brain structures at 2 and 8 weeks after shunting. Because brain glucose levels remained stable from 1 to 12 weeks after shunting, there is no evidence to support the hypothesis that the value of the lumped constant would have changed and caused an artifactual rise in lCMRglc.


Effect of sucrose on chemically and thermally induced unfolding of domain-I of human serum albumin: Solvation dynamics and fluorescence anisotropy study.
The present study is devoted to understand the effect of sucrose on the hydration dynamics and rotational relaxation dynamics within the domain-I of HSA during chemically as well as thermally induced unfolding. It has been observed that the average solvation time become slower in the presence of sucrose for the lower concentrations of GnHCl, however at higher concentrations of GnHCl the effect of sucrose is almost negligible. From the time resolved fluorescence anisotropy it has been observed that in the lower concentration region of GnHCl the sucrose induced stabilization is small as compared to the higher concentrations of GnHCl. We have concluded that the hydration dynamics plays an important role in the sucrose induced stabilization process at the low concentration region; whereas environmental restriction is responsible at the higher concentration of GnHCl. However, we have observed a negligible stabilizing effect of sucrose towards the temperature induced unfolding.


Isolation, comparison of identification methods and antibiotic resistance of Cronobacter spp. in infant foods.
Cronobacter spp. are opportunistic pathogens that cause serious infections, especially in infants, elderly, and immunocompromised people. Dehydrated infant foods are the main vehicle associated with infections caused by these bacteria. Thus, this study aims to investigate the occurrence of Cronobacter spp. in 152 commercial samples of dehydrated infant formulas (77 samples) and dehydrated infant cereals (75 samples), as well as characterize the isolates. Polymerase Chain Reaction (PCR) and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF/MS) methods for isolate identification were used, and their results compared. Furthermore, the susceptibility to 11 antibiotics was tested, and DNA sequencing of one isolate with multi-drug resistance was analyzed. No contamination in the infant formula samples was found, whereas 17.33% (13/75) of the infant cereal samples presented contamination with Cronobacter sakazakii. The identification results by PCR and MALDI-TOF/MS were divergent for some isolates. The antimicrobial resistance results showed a high incidence of resistance to cefazolin (94.4%) besides resistance to amoxicillin (9.45%), cefpodoxime (5.55%), streptomycin (1.35%), and trimethoprim/sulfamethoxazole (1.35%). Whole genome sequencing of one multi-drug resistant isolate showed six genes associated with antimicrobial resistance and an 82% possibility of being a human pathogen based on the presence of virulence factors. The presence of Cronobacter spp. in infant foods represents a risk for the infant's health. Moreover, the presence of a pathogenic multi-drug resistant isolate in infant's food reinforces the necessity of improving food safety policies to protect young children.


Segmentectomy for clinically early-stage primary squamous cell carcinoma of the lung.
Squamous cell carcinoma of the lung-the second most common subtype of lung cancer-has a poorer prognosis than lung adenocarcinoma. However, in contrast to lobectomy, the oncological outcomes after segmentectomy for primary squamous cell carcinomas remain unknown; hence, this study investigated these outcomes. Patients who underwent lobectomy or segmentectomy for clinically node-negative primary lung squamous cell carcinoma with a whole tumor size of ≤ 30 mm on preoperative computed tomography scan during April 2010 to December 2020 were included in this study. The cumulative incidence of recurrence (CIR) among all included patients and propensity score-matched patients were compared using the Gray method. Multivariate analysis using propensity scores and surgical procedures was performed using the Fine and Gray method. Overall, 230 patients were included in this study; of these, 172 (74.8%) underwent lobectomy and 58 (25.2%) underwent segmentectomy. No significant differences were observed in the CIR between patients who underwent lobectomy and those who underwent segmentectomy (5-year rate 18.1% vs. 14.2%; p = 0.787). Moreover, no significant differences in CIR were observed between the propensity score-matched patients who underwent lobectomy (n = 43) and those who underwent segmentectomy (n = 43) (8.6% vs. 8.0%; p = 0.571). Multivariable analysis was performed for CIR using the propensity score; it revealed that segmentectomy was not a significant predictor of worse CIR (hazard ratio, 0.987; p = 0.980). Segmentectomy may be feasible for treating clinically early-stage lung squamous cell carcinoma; its oncological outcomes are similar to those of lobectomy.


Positive and negative effects of diagnostic intensities of ultrasound on erythrocyte blood group markers.
Human erythrocytes were exposed in vitro to diagnostic intensities of ultrasound. Various antigenic tests were conducted to see if the different blood group markers had been changed. No change was observed following sonication for Ss, M, K1, ABO, and the rhesus factors. However a marked reduction was found for the N antigen when assayed using the lectin isolated from Vicia graminea. Several experiments were performed to ensure that this effect was not an artefact. If the cells were not resuspended in fresh medium immediately following sonicating then the magnitude of the effect rapidly decreased. We were unable to detect free N antigen in the supernatants from the sonicated cell suspensions. We were also unable to demonstrate changes in the level of the N antigen following sonication if we used anti-N sera from rabbits. A surprising observation was that different batches of the lectin preparation from the same manufacturer could eliminate the effect.


Perceptions of death, belief systems and the process of coping with chronic pain.
Chronic pain is an on-going experience of embodied discomfort, quite often associated with neuromuscular pathologies, which fails either to heal naturally or to respond to normal medical intervention. The process of coping with chronic pain most commonly involves both the search for medical or non-medical cure, and the search for meaning for intractable suffering. In this paper, I survey various religious, philosophical and mystical belief systems and their empirical use as resources for meaning. The great variability in the ways ideas of death, the key elements extracted from belief systems during the process of coping, are used reflects the variable success in normalizing chronic pain. Theoretically, this paper adds an important dimension to the concept of the chronic illness trajectory, namely, the issue of inevitability, and discusses clinical and non-clinical aspects of depression among people with chronic pain.


Linear vs. circular-stapled gastrojejunostomy in Roux-en-Y gastric bypass.
Various surgical techniques exist to create the gastrojejunostomy during laparoscopic Roux-en-Y gastric bypass (LRYGB). Linear-stapled anastomosis (LSA) and circular-stapled anastomosis (CSA) are two commonly employed techniques. We hypothesized that CSA is associated with an increased rate of surgical site infection (SSI) and gastrojejunostomy stenosis when compared to LSA. This study is a retrospective review of patients who underwent LRYGB for morbid obesity at a single institution between 2012 and 2016. Three bariatric surgeons contributed patients to this series. Clinical information and perioperative outcomes were collected through 90 days after surgery. 171 patients met the inclusion criteria. Two patients did not complete 90-day follow-up and were excluded from the analysis (88 patients CSA, 81 LSA; 99% 90-day follow-up). Patient demographics did not differ between groups. The LSA technique was associated with a significantly reduced rate of SSI (0 (0%) vs. 6 (6.8%), p = 0.02) and stenosis (2 (2.5%) vs. 17 (19.3%), p < 0.01). The CSA technique demonstrated a greater number of endoscopic dilations per stenotic event (1.5 ± 0.8 vs. 1.0 ± 0, p = 0.03). In our experience, a gastrojejunostomy constructed with an LSA technique was associated with a significantly reduced rate of stenosis and SSI compared to the CSA technique. LSA is currently our anastomotic technique of choice in LRYGB.


Photodynamic toxicity of porphyrins and chlorins for a human tumor cell line: combined light and concentration dose responses for the retained fraction.
In recent years porphyrins and related materials have been tested as antitumor agents. A technique was devised to obtain dose-response curves for the sensitizer fraction that resists one day of elution by tissue culture medium--the retained fraction. We found a steep "threshold" dose response relationship that helps to explain tumor destruction without damage to normal tissues. The family of dose-response curves produced by a wide range of light exposures suggests that chlorins and porphyrins do not act by identical mechanisms. Moreover, they suggest that chlorins will prove superior in practical use.


Platelet-activating factor acetylhydrolase is a biomarker of severe anaphylaxis in children.
There is limited ability to predict the severity of allergic reactions in children. Data derived predominantly from adults have implicated the platelet-activating factor pathway as a potential contributor to severe anaphylaxis. In this study, we sought to prospectively assess involvement of key components of the platelet-activating factor pathway in pediatric patients with anaphylaxis. Forty-six pediatric patients (<18 years) presenting with acute anaphylaxis were assessed. Anaphylaxis severity was graded and serum anaphylaxis markers were measured acutely and in 36 children who returned for follow-up >4 weeks after their acute presentation. These markers were compared with pediatric laboratory reference sera. Severe anaphylaxis was experienced by 12/46 (26%) and mild-moderate anaphylaxis in 34/46 (74%) children. Platelet-activating factor acetylhydrolase (PAF-AH) activity was inversely associated with severe anaphylaxis: 9/12 children with severe anaphylaxis had reduced PAF-AH activity as compared with 14/34 with mild-moderate anaphylaxis (p < .05). Furthermore, 3/3 children who required intensive care had markedly reduced mean PAF-AH (nmol/ml/min) (13.73, 95%CI: 7.42-20.03) versus 20/23 who required ward/emergency department care (17.81, 95%CI: 16.80-18.83; p < .05). In children with anaphylaxis, PAF-AH during acute anaphylaxis was unchanged relative to the child's basal levels (mean, 17.26, 95%CI: 16.10-18.42 vs 17.50, 95%CI: 16.21-18.78, p = .63) and was lower than healthy pediatric controls (mean 19.21; 95%CI:18.21-20.21; p < .05). Decreased serum PAF-AH activity is a biomarker of severe anaphylaxis. Levels of this enzyme do not change from basal levels during acute anaphylaxis. Our results show that PAF-AH is a biomarker of anaphylaxis severity in children. This key regulatory enzyme may modulate susceptibility to severe anaphylaxis.


Molecular classification of breast carcinomas with particular emphasis on "basal-like" carcinoma: a critical review.
During the last 11 years, 5 molecular subtypes of breast carcinoma (luminal A, luminal B, Her2-positive, basal-like, and normal breast-like) have been characterized and intensively studied. As genomic research evolves, further subtypes of breast cancers into new "molecular entities" are expected to occur. For example, a new and rare breast cancer subtype, known as claudin-low, has been recently found in human carcinomas and in breast cancer cell lines. There is no doubt that global gene expression analyses using high-throughput biotechnologies have drastically improved our understanding of breast cancer as a heterogeneous disease. The main question is, however, whether new molecular techniques such as gene expression profiling (or signature) should be regarded as the gold standard for identifying breast cancer subtypes. A critical review of the literature clearly shows major problems with current molecular techniques and classification including poor definitions, lack of reproducibility, and lack of quality control. Therefore, the current molecular approaches cannot be incorporated into routine clinical practice and treatment decision making as they are immature or even can be misleading. This review particularly focuses on the "basal-like" breast cancer subtype that represents one of the most popular breast cancer "entities". It critically shows major problems and misconceptions with and about this subtype and challenges the common claim that it represents a "distinct entity". It concludes that the term "basal-like" is misleading and states that there is no evidence that expression of basal-type cytokeratins in a given breast cancer, regardless of other established prognostic factors, does have any impact on clinical outcome.


Recovery of intracellular pH in cortical brain slices following anoxia studied by nuclear magnetic resonance spectroscopy: role of lactate removal, extracellular sodium and sodium/hydrogen exchange.
[31P]- and [1H]nuclear magnetic resonances recorded in an interleaved fashion were used in order to quantify high-energy phosphates, intracellular pH and lactate in cortical brain slices of the guinea-pig superfused in a CO2/HCO3(-)-buffered medium during and after anoxic insults. The volume-averaged intracellular pH and energy status of the preparation following anoxia were determined. In the presence of external Na+, intracellular pH normalized in 3 min and was significantly more alkaline from 10 to 12 min of recovery, but lactate remained elevated for 12 min of reoxygenation following anoxia. The amount of lactate removed was only 40% of the quantity of acid extruded showing operation of H+ neutralizing transmembrane mechanisms other than transport of lactic acid. Amiloride (1 or 2 mM) did not prevent the recovery of intracellular pH, but it blocked the "overshoot" of the alkalinization at 10-12 min of recovery. In a medium containing 70 mM K+, 60 mM Na+ and 0.1 mM Ca2+, the recovery of pH, but not lactate washout, was significantly delayed. Removal of external Na+ caused severe energetic failure, decreases both in oxygen uptake and in N-acetyl aspartate concentration, indicating loss of viable tissue. In Na(+)-free superfusion, lactic acidosis caused a more severe drop in intracellular pH than in the presence of Na+. Complexing of extracellular Ca2+ in the Na(+)-free medium inhibited the acidification by 0.38 pH units during anoxia which is as much as the acidification caused by lactate accumulation in the absence of Na+. In Na(+)-free medium intracellular pH recovered, however, from an anoxic level to a normoxic value in 6 min. Metabolic damage of the slice preparation induced by anoxia in the absence of Na+ was as profound in the presence as in the absence of Ca2+ showing that accumulation of Ca2+ is not the only reason for the damage. It is concluded that recovery of intracellular pH from lactic-acidosis can occur independently of energetic recovery and involves acid extrusion mechanism(s) that is(are) dependent on external Na+ and sensitive to high K+.


Isolated gastric tuberculosis: a case report and review of literature.
Isolated gastric tuberculosis is a rare condition, even in those parts of the world where intestinal tuberculosis is common. Herein we describe a case of a thirty year old female patient with isolated gastric tuberculosis who presented with features of gastric outlet obstruction.


Acrylamide concentration affects the relative position of VP7 gene of serotype G2 strains as determined by polyacrylamide gel electrophoresis.
During the course of development and characterization of various rotavirus reassortants, we found that the relative position of the gene encoding neutralization and protective antigen VP7 of certain rotavirus strains in a PAGE gel was influenced by the concentration of acrylamide. We investigated systematically various factors that affected the relative position of the rotavirus VP7 gene in a PAGE gel. We analyzed dsRNAs of selected rotavirus strains bearing G1, G2, G3 or G9 specificity by PAGE at varying concentrations of acrylamide. We demonstrated that the relative position of the VP7 gene of three G2 strains varied depending upon the concentration of acrylamide in a PAGE gel, which occurred not only in a homologous G2 virus gene background but also in a heterologous G3 virus gene background; and the VP7 gene bearing G1, G3, G4 or G9 specificity did not display this phenomenon when the PAGE running conditions were varied. The concentration of acrylamide in a PAGE gel was the major factor that influenced the relative position of the VP7 gene of G2 rotavirus strains (i.e., VP7 gene coding assignment by PAGE).


isoCNV: in silico optimization of copy number variant detection from targeted or exome sequencing data.
Accurate copy number variant (CNV) detection is especially challenging for both targeted sequencing (TS) and whole-exome sequencing (WES) data. To maximize the performance, the parameters of the CNV calling algorithms should be optimized for each specific dataset. This requires obtaining validated CNV information using either multiplex ligation-dependent probe amplification (MLPA) or array comparative genomic hybridization (aCGH). They are gold standard but time-consuming and costly approaches. We present isoCNV which optimizes the parameters of DECoN algorithm using only NGS data. The parameter optimization process is performed using an in silico CNV validated dataset obtained from the overlapping calls of three algorithms: CNVkit, panelcn.MOPS and DECoN. We evaluated the performance of our tool and showed that increases the sensitivity in both TS and WES real datasets. isoCNV provides an easy-to-use pipeline to optimize DECoN that allows the detection of analysis-ready CNV from a set of DNA alignments obtained under the same conditions. It increases the sensitivity of DECoN without the need for orthogonal methods. isoCNV is available at https://gitlab.com/sequentiateampublic/isocnv .


Cone activation delaying recovery time of rod sensitivity after flash exposure.
The recovery time of rods to the fully dark adapted state after flash exposure was psychophysically and electrophysiologically measured. The recovery time increased abruptly once cones were activated. This abrupt increment was not found for the receptor component (P III) of the frog electroretinogram. It was suggested that the cone activation results in inhibitory influence upon the recovery process of the rod sensitivity.


Computed tomographic diagnosis of ectopic ureter with seminal vesicle cyst.
A rare case of dysplastic kidney with ectopic ureter entering a cystic seminal vesical is reported. This demonstrates the usefulness of computerized tomography in noninvasively delineating such anomalies. Thus accurate preoperative diagnosis can be done, aiding in surgical planning.


Video pneumatic otoscopy for the diagnosis of otitis media with effusion: a quantitative approach.
Pneumatic otoscopy is most helpful for optimally assessing the presence of a middle ear effusion (MEE). To evaluate the diagnostic usefulness and to obtain objective parameters of video pneumatic otoscope (VPO) in ears with MEE, we measured the minimal and maximal pressures in the external auditory canal and recorded the movement of the tympanic membrane (TM) during VPO in 28 ears with MEE and 13 healthy ears. The movements of the TM at static stage, positive and negative pressure stages of VPO were analyzed in terms of the position of the umbo and the angle and length of the malleus. The percent volume of air space out of the total tympanomastoid air cell system (the air index) was obtained from the temporal bone CT, and was used as a reference standard. As a result, minimal pressure and the movement of the umbo from negative pressure stage in ears with MEE were significantly different from the normal ears. The minimal pressure, maximal pressure and the movement of umbo from negative pressure to static stage were significantly correlated with the air index. These results may suggest useful parameters for quantitative analysis of VPO for the visual diagnosis of MEE.


Ad-libitum fluid intake was insufficient to achieve euhydration 20 h after intermittent running in male team sports athletes.
This study documented 20 h rehydration from intermittent running while concealing the primary outcome of rehydration from subjects. Twenty-eight male team sports athletes (age 25 ± 3 y; predicted V̇O2max 54 ± 3 mL kg-1 min-1) were pair-matched to exercise (EX) or rest (REST) groups. To determine hydration status, body mass, urine and blood samples were collected at 08:00, pre-intervention (09:30), post-intervention (12:00), 3 h post-intervention and 08:00 the following morning (20 h). The intervention was 110 min intermittent running (EX) or seated rest (REST), with ad-libitum fluid provided in both. Subjects completed a weighed diet record and collected all urine for the 24 h. Changes typical of hypohydration were apparent in EX following the intervention period (body mass: EX -2.0 ± 0.5%; REST -0.2 ± 0.3%; serum osmolality: EX 293 ± 4 mOsm∙kgH2O-1; REST 287 ± 6 mOsm∙kgH2O-1; P ≤ 0.022). Fluid intake during the intervention period (EX 704 ± 286 mL, REST 343 ± 230 mL) and fluid intake within the first 3 h post-intervention (EX 1081 ± 460 mL, REST 662 ± 230 mL) were greater (P ≤ 0.004), and 24 h urine volume lower (EX 1697 ± 824 mL, REST 2370 ± 842 mL; P = 0.039) in EX. Compared to baseline, body mass remained lower (-0.6 ± 0.5%; P = 0.030) and urine osmolality elevated (20 h: 844 ± 197 mOsm∙kgH2O-1, 08:00: 698 ± 200 mOsm∙kgH2O-1; P = 0.004) at 20 h in EX. When games players drank fluid ad-libitum during exercise and post-exercise in free-living conditions, a small degree of hypohydration remained 20 h post-exercise.


A mechanism-based mathematical model of aryl hydrocarbon receptor-mediated CYP1A induction in rats using beta-naphthoflavone as a tool compound.
β-Naphthoflavone (BNF) is a synthetic flavone that selectively and potently induces CYP1A enzymes via aryl hydrocarbon receptor activation. Mechanism-based mathematical models of CYP1A enzyme induction were developed to predict the time course of enzyme induction and quantitatively evaluate the interrelationship between BNF plasma concentrations, hepatic CYP1A1 and CYP1A2 mRNA levels, and CYP1A enzyme activity in rats in vivo. Male Sprague-Dawley rats received a continuous intravenous infusion of vehicle or 1.5 or 6 mg · kg(-1) · h(-1) BNF for 6 h, with blood and liver sampling. Plasma BNF concentrations were determined by liquid chromatography-tandem mass spectrometry. Hepatic mRNA levels of CYP1A1 and CYP1A2 were determined by TaqMan. Ethoxyresorufin O-deethylation was used to measure the increase in CYP1A enzyme activity as a result of induction. The induction of hepatic CYP1A1/CYP1A2 mRNA and CYP1A activity occurred within 2 h after BNF administration. This caused a rapid increase in metabolic clearance of BNF, resulting in plasma concentrations declining during the infusion. Overall, the enzyme induction models developed in this study adequately captured the time course of BNF pharmacokinetics, CYP1A1/CYP1A2 mRNA levels, and increases in CYP1A enzyme activity data for both dose groups simultaneously. The model-predicted degradation half-life of CYP1A enzyme activity is comparable with previously reported values. The present results also confirm a previous in vitro finding that CYP1A1 is the predominant contributor to CYP1A induction. These physiologically based models provide a basis for predicting drug-induced toxicity in humans from in vitro and preclinical data and can be a valuable tool in drug development.


Our Commitment to Promoting Diversity, Equity, Inclusion, and Justice in Child Maltreatment.
In this commentary, the editorial team of Child Maltreatment extends and expands on APSAC's position on diversity, equity, inclusion, and justice, affirms our commitment and plans for addressing these issues in this publication, and highlights articles in this issue that continue the discussion about race and racism in the child welfare and child protection systems.


A randomized controlled trial of an mHealth intervention for increasing access to HIV testing and care among young cisgender men and transgender women: the mLab App study protocol.
The number of youth living with HIV in the United States (US) continues to rise, and racial, ethnic, and sexual minority youth including young men who have sex with men (YMSM) and young transgender women (YTGW) bear a disproportionate burden of the HIV epidemic. Due to social and healthcare system factors, many YMSM and YTGW do not seek HIV testing services and are therefore less likely to be aware that they are infected. Mobile health technology (mHealth) has the ability to increase uptake of HIV testing among these populations. Thus, the mLab App-which combines HIV prevention information with a mobile phone imaging feature for interpreting at-home HIV test results-was developed to improve testing rates and linkage to care among Black, Latino, and other YMSM and YTGW living in New York City and Chicago and their surrounding areas. This study is a three-arm randomized controlled trial among YMSM and YTGW aged 18-29 years. Participants are randomized to either the mLab App intervention including HIV home test kits and standard of preventive care, standard of preventive care only, or HIV home test kits and standard of preventive care only. mHealth technology used for HIV prevention is capable of delivering interventions in real-time, which creates an opportunity to remotely reach users across the country to strengthen their HIV care continuum engagement and treatment outcomes. Specifically during the COVID-19 pandemic, mHealth technology combined with at-home testing may prove to be essential in increasing HIV testing rates, especially among populations at high-risk or without regular access to HIV testing. This trial was registered with Clinicaltrials.gov ( NCT03803683 ) on January 14, 2019.


Rods of actin filaments in type I hair cells of the Shaker-2 mouse.
The shaker-2 mouse with inherited inner ear disease suffers from deafness and a shaking-waltzing behavior. The hair cell type I in cristae ampullares and maculae utriculi show a specific pathology, featuring fusion of the stereocilia and presence of a rod-shaped inclusion body. The inclusion body is composed of filaments that could be identified as the protein actin by the method of decoration with subfragment S-1 of myosin. The functional polarity was determined, and S-1 fragments were found to point apically, that is, from the nucleus up toward the cuticular plate. These observations are identical to those earlier described in the waltzing guinea pig. It is concluded that the identical pathology at a cellular level in two different species may indicate a pathologic disorder in a process fundamental to the normal development of this type of hair cell.


Effects of zinc oxide-eugenol on leucocyte number and lipoxygenase products in artificially inflamed rat dental pulp.
The number of leucocytes and level of endogenous LTC4 in the pulp tissue were measured by a histological method and radioimmunoassay, respectively. When the mandibular incisor pulp was irritated by drilling a hole in the dentine without using any coolant, the number of polymorphonuclear leucocytes and lymphocytes and the concentration of LTC4 increased to 3.2, 1.9 and 1.8 times their respective levels in normal pulp 6 h after the injury. The total leucocyte number in blood collected from these rats was also increased significantly. In contrast, when the cavities cut in dentine were filled with a zinc oxide-eugenol mixture (powder: liquid 100 mg/25 microliters), the increase in the number of cells was significantly curtailed, and the LTC4 level fell to 50% of that in normal pulp within 1 h after the filling. No decrease in the LTC4 level was observed after filling with a zinc oxide-water mixture, but the level decreased in response to an increase in eugenol content in the zinc oxide-eugenol placed in the cavity. Biosynthesis of [14C]-HETE and HPETE from [14C]-arachidonic acid was inhibited by the addition of 10 microM eugenol to the pulp homogenate. Thus eugenol released from zinc oxide-eugenol inhibited the biosynthesis of lipoxygenase products and the early chemotactic accumulation of leucocytes.


Enhanced recovery after surgery (ERAS) pathway vs traditional care in laparoscopic rectal resection: a single-center experience.
The aim of this study was to compare the outcome of an enhanced recovery after surgery (ERAS) pathway with traditional perioperative care in laparoscopic rectal resection. A retrospective analysis of prospectively collected data was conducted. Single-center consecutive patients who underwent laparoscopic rectal surgery after an ERAS program were compared with patients who received traditional care over an 8-year period. Primary and total length of stay, and readmission, morbidity and mortality rates were analyzed. For ERAS group, the actual adherence to protocol was also evaluated. Two hundred and ninety-seven patients, 162 in the ERAS group and 135 in conventional care, were studied. Median primary and total length of stay were significantly shorter in the ERAS group (9 vs 12 days; p = 0.0001; 10 vs 12 days; p = 0.01; respectively). The ERAS group experienced a faster recovery of bowel function than the traditional care group (p = 0.0001). A similar morbidity rate was observed in the two groups (32.3 % in ERAS vs 36.1 % in traditional care p = 0.41). Readmission rates were 4.9 % in the ERAS versus 1.5 % in the traditional care group (p = 0.19). There was no mortality in either group. Overall mean compliance with the ERAS protocol was 85.7 % (range 54.4-100 %). The introduction of the ERAS protocol in laparoscopic rectal resection led to a reduction in primary and total length of hospital stay without an increase in morbidity or readmission rates when compared to traditional care.


Operative Distal Humerus Fractures in Older Patients: Predictors for Early Complications Based on a National Database.
Open reduction internal fixation (ORIF) and total elbow arthroplasty (TEA) have both been utilized in the treatment of distal humerus fractures in patients over 65 years of age. Comparisons of early complications between these procedures have not been well described. The purpose of this study is to evaluate complication rates in the treatment of distal humerus fractures in elderly patients and to utilize prediction models to identify risk factors associated with postoperative complications. Additionally, to compare ORIF and TEA treatment. A retrospective case series was performed by querying the National Surgical Quality Improvement Program for both ORIF and TEA performed for distal humerus fractures in patients over the age of 65 years between 2005 and 2014. We examined both preoperative risk factors and complications within 30 days in these three groups. The sample included 216 ORIF and 65 TEA cases. No outcomes examined differed significantly between treatment groups. The most common outcome for both groups was bleeding requiring transfusion (8%). The observed effect size for the association between procedure and the composite morbidity outcome indicated little to no association (phi = 0.004). Furthermore, no presurgery variables were found to be significantly associated with procedure type. The only predictor with a significant independent association with the composite outcome, regardless of procedure type, was ASA class 3/4. Clinical complications were low no matter the type of treatment. Low preoperative hematocrit was a risk factor in both ORIF and TEA.


Studying RNA Polymerase II Promoter-Proximal Pausing by In Vitro Immobilized Template and Transcription Assays.
The immobilized template assay is a versatile biochemical method for studying protein-nucleic acid interactions. Using this method, immobilized nucleic acid-associated or specific proteins can be identified and quantified by techniques such as mass spectrometry and immunoblotting. Here, a modified immobilized template assay combined with in vitro transcription assay to study the function of transcription factors and transcriptional activities at the human heat shock protein 70 (HSP70) gene is described. Notably, this method can be applied to study other important genes and transcription factors in vitro.


An improved authentication scheme for telecare medicine information systems.
The telecare medicine information system enables or supports health-care delivery services. In order to safeguard patients' privacy, such as telephone number, medical record number, health information, etc., a secure authentication scheme will thus be in demand. Recently, Wu et al. proposed a smart card based password authentication scheme for the telecare medicine information system. Later, He et al. pointed out that Wu et al.'s scheme could not resist impersonation attacks and insider attacks, and then presented a new scheme. In this paper, we show that both of them fail to achieve two-factor authentication as smart card based password authentication schemes should achieve. We also propose an improved authentication scheme for the telecare medicine information system, and demonstrate that the improved one satisfies the security requirements of two-factor authentication and is also efficient.


Progressive Multiple Sclerosis.
Many therapeutic advances for relapsing-remitting multiple sclerosis (MS) have occurred in the past 25 years. Although similar advances in disease-modifying therapies have not been realized in progressive MS, many symptomatic therapeutic strategies can benefit patients with progressive MS. Few guidelines exist for management of patients with progressive MS. The classification of progressive MS was revised in 2013 to include a description of inflammatory disease activity determined by clinical relapses or imaging findings. Developing knowledge about the pathogenesis of progressive MS and the role of comorbidities in modifying the disease course has implications for the clinical management of patients with progressive MS as well as for clinical trial design. Current and upcoming clinical trials will assess a wide range of interventions, including immunomodulatory agents, putative neuroprotective molecules, stem cell therapy, nutrition, and rehabilitation techniques. None of the therapies currently approved for use in relapsing-remitting MS have been shown to slow the gradual progression of disability that occurs in the absence of recent relapses or changes in MRI. A multidisciplinary approach is needed to address the many symptoms that impact quality of life for patients with progressive MS.


Amyloidosis: a clinical overview.
Amyloidosis is the name for protein-folding diseases characterized by extracellular deposition of a specific soluble precursor protein that aggregates in the form of insoluble fibrils. The classification of amyloidosis is based on the chemical characterization of the precursor protein. Deposition of amyloid is localized or systemic. The 4 main types of systemic amyloidosis are AL, AA, ATTR, and Aβ2M type. A schematic approach is proposed for the clinical management of systemic amyloidosis. The importance of typing amyloid with confidence, the usefulness of imaging techniques, the principles of treatment, and the need for well-planned treatment monitoring during follow-up are discussed.


Suprapubic catheter insertion using an ultrasound-guided technique and literature review.
What's known on the subject? and What does the study add? The conventional 'blind' technique for suprapubic catheter (SPC) insertion relies on adequate filling of the bladder to displace bowel away from the site of needle puncture. However, in a small percentage of patients this fails to happen, which can occasionally lead to life-threatening bowel injury. Recently published British Association of Urological Surgeons (BAUS) guidelines have recommended that ultrasonography (US) may be helpful to identify bowel loops and recommends its usage whenever possible. This paper describes the technique of US-guided needle puncture and SPC insertion to reduce the likelihood of bowel injury. The paper addresses training, equipment and logistical issues associated with this advice. We have reviewed the available publications on the outcomes from this technique and also present our experience. Suprapubic catheter (SPC) insertion is a common method of bladder drainage in contemporary urological practice. The procedure involves insertion of a sharp trocar into the bladder percutaneously, usually by palpation, percussion or cystoscopy for guidance. Although generally considered a safe procedure, the risk of bowel injury is estimated at up to 2.4% with a mortality rate of 1.8%. Recently published British Association of Urological Surgeons (BAUS) guidelines have recommended that ultrasonography (US) may be helpful to identify bowel loops and recommends its usage whenever possible. The present paper describes the use of US for SPC insertion and discusses the implications of this advice. This paper is designed to support and supplement practical techniques learnt on a course and in clinical practice.


[Acute heart insufficiency in an 8-month-old infant presenting with hypocalcemia and Epstein-Barr virus infection: acute myocarditis? Or primary hypokinetic dilated cardiomyopathy?].
An eight-month-old was admitted for acute congestive heart failure with fever. The respective parts played by hypocalcemia (due to vitamin-D deficiency rickets) and acute Epstein-Barr virus infection are discussed. Hypocalcemia was sufficiently marked to induce heart failure per se but replenishment of calcium stores was followed by only partial improvement in cardiac manifestations. Initial management was difficult because of the risks associated with concomitant administration of calcium and digitalis. After eighteen months during which the patient's status remained stable, evaluation showed that clinical features were consistent with sequelae of acute viral myocarditis. The possibility of primary hypokinetic dilated cardiomyopathy was then considered. Esterified carnitine levels were found to be increased leading to further investigations which outruled mitochondrial cytopathy.


Brain lesions and water-maze learning deficits after systemic administration of kainic acid to adult rats.
The relationship between hippocampal damage and spatial learning deficiencies was studied in rats injected with kainic acid (10 mg/kg i.p.). A single injection was given either before or after the acquisition phase of the Morris water-maze task. In this acquisition phase, the animals were required to find a hidden underwater platform starting from four different points. The task was repeated twice a day for 10 days. In the retention phase after 10 days rest, the rats repeated the same task. The damage caused by the treatment occurred in several prosencephalic areas, including the piriform and enthorhinal cortices, the thalamus and the hippocampus. In the latter, greatest damage was seen in CA1 followed by CA3 while CA2 and the gyrus dentatus appeared almost unaffected. The behavioural results indicated that kainic acid impaired but did not preclude the acquisition of the water-maze task. During the retention phase, no significant differences in latencies were found between animals that were treated before and after acquisition, thus, indicating that pretraining does not play an important role in the recovery of these spatial abilities following hippocampal lesions.


Antibodies to intermediate filament proteins as molecular markers in clinical tumor pathology. Differentiation of carcinomas by their reaction with different cytokeratin antibodies.
Antibodies to human and bovine epidermal prekeratin and antibodies to mouse liver cytokeratin component D (Mr 49 000) have been applied in indirect immunofluorescence microscopy on sections of human tumors of mammary gland and liver. In non-neoplastic mammary gland all epithelial cells were stained with these antibodies. In pre-invasive and invasive ductal and lobular carcinomas a cell population was observed which was not significantly stained with antibodies to epidermal prekeratin but did strongly react with antibodies to liver cytokeratin D. In the liver, the antibodies to epidermal prekeratin as well as those directed against liver cytokeratin D strongly decorated bile duct epithelia. In contrast, significant staining of the hepatocytes was only achieved with antibodies to liver cytokeratin D. This different staining reaction was maintained in liver tumors of hepatocellular and cholangiocellular origin. Antibodies to vimentin stained mesenchymal cells and tumors of mesenchymal derivation but reacted not significantly with any of the epithelial and carcinoma cells examined. The difference is of practical importance for the discrimination between anaplastic carcinomas and sarcomas of unknown origin. Cytokeratin could also be detected by antibody staining using the peroxidase-antiperoxidase (PAP) technique in formaldehyde-fixed and paraffin-embedded material of skin, gastrointestinal, respiratory, urinary and genital tract as well as various glands, liver and kidney. Examples of positive reactions were shown in a squamous cell carcinoma, a basalioma and a pleomorphic adenoma of the parotis. It is concluded that the immunohistochemical analysis of intermediate filament proteins has diagnostic potential in clinical pathology and may help to elucidate histogenesis and differentiation of tumors and possibly also prognosis of tumor growth. It is further suggested to use antibodies recognizing different subsets of proteins of the cytokeratin family in order to distinguish between different types of carcinomas.


[Lateral Intercostal Artery Perforator (LICAP) flap for lateral breast defect reconstruction].
Lateral breast defect after partial mastectomy is a difficult challenge. Pedicled perforator flap is a new indication for breast surgery. Perforator flaps and fat tissue transplant are new concepts in this kind of reconstruction. Lateral intercostal artery perforator (LICAP) flap has been used for a lot of indications. It can be used for lateral breast defect reconstruction. This flap provides several advantages: minimal donor site morbidity, advantages in flap shaping, better aesthetic results and higher patient satisfaction. The aim of this paper is to present indications of LICAP flap. We discuss its advantages and its limits in lateral breast reconstruction after partial mastectomy. LICAP flap provides a new solution instead of conventional flaps.


miR-214-5p targets KLF5 and suppresses proliferation of human hepatocellular carcinoma cells.
MicroRNAs (miRNAs) are small endogenous conserved RNAs regulating genes expression through base pairing with the 3'-untranslated region (3'-UTR) of target messenger RNAs. MiR-214-5p is a newly identified miRNA with its biological role largely unknown. In this study, we explored miR-214-5p expression status in 78 paired tumor and nontumor tissues obtained from patients with hepatocellular carcinoma (HCC) by RT-qPCR. The effects of miR-214-5p expression on HCC cell proliferation, cell cycle progression, and cell migration were measured by CCK-8 assay, flow cytometry, and wound-healing assay. A dual-luciferase activity assay was performed to identify whether KLF5 was a target of miR-214-5p. Kaplan-Meier curve and log-rank test were used to investigate the effects of miR-214-5p and KLF5 on overall survival and disease-free survival of patients with HCC. We found miR-214-5p expression was sharply reduced in HCC tissues and cell lines compared with the normal tissues and cell lines. Functional assay revealed that miR-214-5p overexpression could downregulate cell proliferation, cell migration, and arrested cell cycle at G0/G1 phase. Further, we validated Krüppel-like factor 5 (KLF5) as a direct target of miR-214-5p, and was upregulated in HCC and inversely correlated with the expression of miR-214-5p. Moreover, we found the low expression of miR-214-5p and high expression of KLF5 were correlated with tumor size, tumor stage, and poorer 5-year overall survival and disease-free survival of patients with HCC. In conclusion, our results suggested miR-214-5p functions as a tumor suppressor through targeting KLF5 in HCC. Also, miR-214-5p and KLF5 were identified as potential prognostic markers and might be therapeutic targets in HCC.


Downregulation of ferroportin 1 expression in hFOB1.19 osteoblasts by hepcidin.
The present study was designed to address the relationship between iron homeostasis and bone metabolism. Cultured hFOB 1.19 osteoblasts were incubated with selected concentrations of hepcidin (50, 100, and 200 nmol/L) for 20 h, harvested for extraction of total RNA and proteins, and the expression of ferroportin 1 was analyzed by RT-PCR and western blotting. The results showed the presence of ferroportin 1 expression in cultured hFOB 1.19 cells. Furthermore, the ferroportin 1 had a similar expression pattern in hFOB cells as in hepatocytes and enterocytes and was downregulated by hepcidin. Our data indicate that osteoblasts are target cells for hepcidin, suggest that hepcidin may have many more targets than previously recognized, and support the role of hepcidin in the development of osteoporosis.


Clouding phenomenon and SANS studies on tetra-n-butylammonium dodecylsulfate micellar solutions in the absence and presence of salts.
Clouding phenomenon in aqueous micellar solutions of an anionic surfactant tetra-n-butylammonium dodecylsulfate (TBADS) has been observed as a function of surfactant concentration. Small-angle neutron scattering (SANS) experiments in these systems show clustering of micelles as the temperature approaches the cloud point (CP). The individual micelles and the clusters of micelles coexist at CP. The clustering of micelles depends on the surfactant concentration and temperature. It is proposed that clustering is due to depletion of H-bonded water present around the butyl chains at the micellar surface. This is associated with entropy gain which is considered to be the major thermodynamic factor related to micellar aggregation. The structures (clusters) that emerge depend on the relative lengths of the alkyl chains of the counterion and can be tuned by the temperature.


Phylogeography and biogeography of fungi.
The rigorous study of processes shaping geographic distributions of lineages is a relatively new and emerging field in mycology. While it was previously generally believed that most fungi have wide distributions and largely unstructured populations, recent studies have shown that this is not the case. The study of distributions in tandem with molecular approaches to phylogeny has recently made substantial advances to our understanding of the diversity and biogeography of fungi. Comprehensive species inventories have provided a better picture of the actual distribution of these organisms, while robust phylogenies based on molecular characters have provided both data that allow interpretation of current distributions and testable hypotheses regarding the processes responsible for distribution patterns. This commentary provides an introduction to five papers in this issue of Mycological Research that focus on fungal phylogeography. These papers are based on oral contributions given at two symposia at the International Mycological Congress (IMC8) held in Cairns (Australia) in August 2006.


Comparisons of exacerbations and mortality among LAMA/LABA combinations in stable chronic obstructive pulmonary disease: systematic review and Bayesian network meta-analysis.
Only few randomized controlled trials (RCTs) for head-to-head comparison have been conducted between various combinations of long-acting muscarinic antagonists (LAMAs) and long-acting beta-agonists (LABAs). Our study was conducted to compare acute exacerbation and all-cause mortality among different LAMA/LABA regimens using Bayesian network meta-analysis (NMA). We searched Medline, EMBASE, and the Cochrane library (search date: July 1, 2019). We included parallel-group RCTs comparing LAMA/LABA combinations with other inhaled drugs in the stable COPD for ≥ 48 weeks. Two different network geometries were used. The geometry of network (A) had nodes of individual drugs or their combination, while that of network (B) combined all other treatments except LAMA/LABA into each drug class. This study was prospectively registered in PROSPERO; CRD42019126753. We included 16 RCTs involving a total of 39,065 patients with stable COPD. Six combinations of LAMA/LABA were identified: tiotropium/salmeterol, glycopyrrolate/indacaterol, umeclidinium/vilanterol, tiotropium/olodaterol, aclidinium/formoterol, and glycopyrrolate/formoterol. We found that umeclidinium/vilanterol was associated with a lower risk of total exacerbations than other LAMA/LABAs in the NMA using network (A) (level of evidence: low or moderate). However, the significant differences were not present in the NMA of network (B). There were no significant differences among the LAMA/LABA combinations in terms of the number of moderate to severe exacerbations, all-cause mortality, major adverse cardiovascular events, or pneumonia. The present NMA including all available RCTs provided that there is no strong evidence suggesting different benefits among LAMA/LABAs in patients with stable COPD who have been followed up for 48 weeks or more. This study was prospectively registered in PROSPERO; CRD42019126753.


Transcorporal artificial urinary sphincter in radiated and non - radiated compromised urethra. Assessment with a minimum 2 year follow-up.
to assess the efficacy of transcorporal artificial urinary sphincter (AUS) implantation on continence for male stress urinary incontinence in cases of prior surgical treatment or/and radiation failure, and as a first option in radiation patients. From March 2007 to August 2012, 37 male patients were treated with transcorporal AUS AMS™ 800. Twelve patients had primary placement of transcorporal cuff, a surgical option due to a previous history of radiation and 25 patients had secondary procedure after failure of AUS or urinary incontinence surgery. Functional urinary outcomes were assessed by daily pad use, 24-hour Pad-test and ICIQ-SF questionnaire. Quality of life and satisfaction were assessed based on I-QoL and PGI-I questionnaires. After a median of 32 months, the continence rate (0 to 1 pad) was 69.7%. Median pad test was 17.5g (0-159), mean ICIQ-SF score was 7.3/21 (±5.4) and mean I-QoL score was 93.9/110. A total of 88% of the patients reported satisfaction with the AUS. The 5-year actuarial revision-free for AUS total device was 51%. Patients for primary implant for radiation were not more likely to experience revision than non-radiation patients. Preservation of erections was reported in half of the potent patients. Transcorporal AUS cuff placement is a useful alternative procedure option for severe male UI treatment, especially in patients with a compromised urethra after prior surgery or radiation. A high continence rate was reported and implantation as first option in radiation patients should be considered.


Isotope effect studies on the dehydrogenation of delta 1-tetrahydrocannabinol in the rat.
Isotope effect studies on the metabolic dehydrogenation of delta 1-tetrahydrocannabinol in rats are described and it is shown that this process is confined to a very short period following i.v. administration. The implications of this finding are discussed.


Relationship between arterial potassium and ventilation during exercise in patients with chronic heart failure.
The mechanisms underlying the increased ventilatory response to exercise seen in patients with chronic heart failure are not clearly understood. Arterial potassium has been suggested as an important ventilatory stimulant. The authors have investigated the arterial potassium response in patients with heart failure. Although arterial potassium rises during exercise, no evidence was found to suggest a greater potassium response in patients with heart failure compared to normal subjects. There was no direct correlation between the rise in ventilation and the rise in arterial potassium. It remains possible that there is an increased sensitivity to arterial potassium in patients with heart failure, but it would need to be three times greater than in normal subjects.


Chitosan based edible films and coatings: a review.
Chitosan is a biodegradable biocompatible polymer derived from natural renewable resources with numerous applications in various fields, and one of which is the area of edible films and coatings. Chitosan has antibacterial and antifungal properties which qualify it for food protection, however, its weak mechanical properties, gas and water vapor permeability limit its uses. This review discusses the application of chitosan and its blends with other natural polymers such as starch and other ingredients for example essential oils, and clay in the field of edible films for food protection. The mechanical behavior and the gas and water vapor permeability of the films are also discussed. References dealing with the antimicrobial behavior of these films and their impact on food protection are explored.


Scientific counterfactuals as make-believe.
Counterfactuals abound in science, especially when reasoning about and with models. This often requires entertaining counterfactual conditionals with nomologically or metaphysically impossible antecedents, namely, counternomics or counterpossibles. In this paper I defend the make-believe view of scientific counterfactuals, a naturalised fiction-based account of counterfactuals in science which provides a means to evaluate their meanings independently of the possibility of the states of affairs their antecedents describe, and under which they have non-trivial truth-values. Fiction is here understood as imagination (in contrast with its most typical association with falsity), characterised as a propositional attitude of pretense or 'make-believe' (Walton 1990). The application of this theory to scientific counterfactuals makes their evaluation a game of make-believe: a counterfactual is (fictionally) true iff its antecedent and the rules of the game prescribe the imagining of its consequent (Kimpton-Nye 2020). The result is a practice-based account of counterfactuals and counterfactual reasoning in science which incorporates insights from theoretical and experimental analytic philosophy as well as cognitive science. This way, the make-believe view of scientific counterfactuals shows that the evaluation of scientific counterfactuals is none other than a question of scientific representation in disguise.


Safety and impact of donor-type red blood cell transfusion before allogeneic peripheral blood progenitor cell transplantation with major ABO mismatch.
Changes within the ABO system are regularly observed phenomena in allogeneic bone marrow transplantation (BMT) and peripheral blood progenitor cell transplantation (PBPCT). Major ABO mismatch can lead to different clinical problems including acute hemolysis after infusion of the allograft, delay of red blood cell (RBC) engraftment, or even manifestation of pure red cell aplasia (PRCA). This retrospective study demonstrates the safety and the impact of donor-type RBC transfusion before allogeneic PBPCT in major ABO settings as routinely performed at our transplantation unit. This study reports on transfusion of mismatched RBCs at the end of the conditioning period in 35 patients who underwent allogeneic PBPCT, which led to a decrease in isoagglutinin titers in most cases. A decrease of isoagglutinin titer after donor-type RBC transfusion can significantly reduce the demand of RBC transfusion between transplantation and Day +30 (p = 0.003). Interestingly, patients who developed PRCA were not observed, a complication being regularly documented by other groups. A decrease of isoagglutinin titers by in vivo immunoadsorption before allogeneic PBPCT does not only lack severe complication but also leads to a reduction in demand of RBC transfusion after engraftment and may reduce the incidence of PRCA in these patients.


Plasma 25-hydroxyvitamin D and risk of premenstrual syndrome in a prospective cohort study.
Moderate to severe premenstrual syndrome (PMS) affects 8-20 percent of premenopausal women. Previous studies suggest that high dietary vitamin D intake may reduce risk. However, vitamin D status is influenced by both dietary vitamin D intake and sunlight exposure and the association of vitamin D status with PMS remains unclear. We assessed the relation of plasma 25-hydroxyvitamin D (25OHD), total calcium and parathyroid hormone levels with risk of PMS and specific menstrual symptoms in a case-control study nested within the prospective Nurses' Health Study II. Cases were 401 women free from PMS at baseline who developed PMS during follow-up (1991-2005). Controls were women not experiencing PMS (1991-2005), matched 1:1 with cases on age and other factors. Timed luteal phase blood samples were collected between 1996 and 1999 from cases and controls. We used conditional logistic regression to model the relation of 25OHD levels with risk of PMS and individual menstrual symptoms. In analyses of all cases and controls, 25OHD levels were not associated with risk of PMS. However, results differed when the timing of blood collection vs. PMS diagnosis was considered. Among cases who had already been diagnosed with PMS at the time of blood collection (n = 279), 25OHD levels were positively associated with PMS, with each 10 nmol/L change in 25OHD associated with a 13% higher risk. Among cases who developed PMS after blood collection (n = 123), 25OHD levels were unrelated to risk of PMS overall, but inversely related to risk of specific menstrual symptoms. For example, each 10 nmol/L increase was associated with a significant 21% lower risk of breast tenderness (P = 0.02). Total calcium or parathyroid hormone levels were unrelated to PMS. 25OHD levels were not associated with overall risk of PMS. The positive association observed among women already experiencing PMS at the time of 25OHD measurement is likely due to confounding by indication related to use of dietary supplements to treat menstrual symptoms. Results from prospective analyses, which were less likely influenced by this bias, suggest that higher 25OHD levels may be inversely related to the development of specific menstrual symptoms.


Predicting pilot's sleep during layovers using their own behaviour or data from colleagues: implications for biomathematical models.
Biomathematical models are used in industry to estimate how much sleep people are likely to get on different work patterns, and how efficient and safe people are likely to be at work. Since there is evidence to suggest that individuals respond differently to sleep loss, there has been a recent focus on trying to account for individual differences. One possible approach could use past behaviour to predict future responses to similar working conditions. This study investigated the predictive value of sleep timing and duration data for a particular individual on a break between shifts relative to data from their colleagues. Sleep diaries and wrist actigraphy were collected from 306 international long-haul pilots for at least 2-weeks. Fifty layovers, equivalent in origin and destination, length and timing, were completed twice by individual pilots. Matched layovers done by other pilots (n=2311) were also identified. Layover periods were analysed for minute-by-minute correspondence of sleep or wake (yes/no), and total sleep time (TST). Using an individual's own data improved concordance by approximately 5% relative to using a large sample of different pilots, and by 10% relative to using a random sample of 50 different pilots. Using an individual's own TST to predict their TST on an equivalent layover yielded an r value of 0.83, compared to r=0.78 when data from a colleague was used, and r=0.73 using different pilots in a random sample of equivalent size. The mean difference in TST using pilots' own data was <20 min, compared to <40 min using data from colleagues. However, the confidence limits on these differences were large (up to 8h). Results suggest that for international pilots on specific layover patterns, knowing the past behaviour of an individual may only represent a modest improvement over knowing the length and timing of a colleague's sleep, when it comes to predicting their sleep behaviour.


Diagnostic yield and complications of extended lymphadenectomy versus limited lymphadenectomy combined with radical prostatectomy.
Lymphadenectomy for prostate cancer (PC) is the most reliable procedure for detecting lymphatic metastases. The optimal extension of this procedure is still a topic of debate. To analyse the diagnostic performance and complications of extended lymphadenectomy (ELD) and limited lymphadenectomy (LLD) in a series of patients with high-risk PC who underwent radical prostatectomy (RP). A retrospective study was conducted on patients with high d'Amico risk who underwent RP with lymphadenectomy between 1999 and 2014. A comparative analysis was performed of the diagnostic capacity of lymphatic metastases of ELD and LLD and of postoperative complications at 90 days. Ninety-three patients were analysed, 20 (21.5%) and 73 (78.5%) of whom underwent ELD and LLD, respectively. The mean age of the series was 65.26 years (SD, 5.51). The median follow-up was 1.51 (0.61-2.29) years in the ELD group and 5.94 (3.61-9.10) in the LLD group. The median number of nodes obtained was 13 (9-23) in the ELD group compared with 5 (2-8) in the LLD group (p <.001). The percentages of patients with positive nodes in the ELD and LLD groups were 35% and 5.47%, respectively (p <.001). The overall complication rate at 90 days was 35.5% (33 patients). In the ELD group, 12 patients (60%) had complications, compared with 21 patients (28.8%) in the LLD group (p=.016), with no significant differences in severity according to the Clavien scale (p=.73). In our series, the detection of metastatic nodes was significantly greater with ELD. ELD increases the number of complications, with no differences compared with LLD in severity according to the modified Clavien scale.


The social versus food preference test: A behavioral paradigm for studying competing motivated behaviors in rodents.
Behavior is influenced by a combination of factors, with the expression of the appropriate behavior dependent on an individual's current motivational state and the presence of stimuli in their surrounding environment. Thus far, most laboratory studies have focused on uncovering the peripheral and central systems that regulate the expression of a single behavior or the expression of a suite of behaviors associated with a single motivational state. In natural settings, however, an individual can be simultaneously experiencing multiple motivational states with multiple choices of how to act. Yet, the direct assessment of the roles of peripheral and central systems in coordinating motivated behavioral choice is largely understudied. This may be due to a lack of behavioral tests that are suitable for such investigations. Here, we describe a recently developed behavioral paradigm, hereafter called the Social versus Food Preference Test. This behavioral paradigm was validated in both rats and mice and is highly flexible, which will allow addressing of a wide range of research questions concerning the peripheral and central systems that coordinate the choice to seek social interaction versus the choice to seek food.•This paradigm was validated in rats and mice, the two most commonly used nonhuman species in behavioral research, but could be adapted for use in other rodent models.•The specific social and food stimuli used can be selected based on the research question.•Three-chamber apparatuses can be custom-constructed.


Heavy metal blood levels and hearing loss in children of West Bengal, India.
Heavy metals are a major environmental threat in India and there are several health risks associated with it. The aim of this study was to investigate the relationship between the blood levels of lead, cadmium, arsenic, and mercury and a sensoneurial hearing loss in children aged one to ten years. Heavy metal blood levels were determined using inductively coupled plasma mass spectrometry, with appropriate quality control. We found significantly higher blood lead concentration (mg/L; Mean ± SE) in children with a hearing loss (53.2 ± 4.4) compared to healthy controls (38.4 ± 4.7)/P = 0 0.03/. Children's blood lead levels ≥ 50 mg/L compared to the levels < 10 mg/L were associated with increased probability of hearing loss (OR, 48.8; 95% CI, 41.9-55.6). The differences in the blood levels of cadmium, arsenic, and mercury between the children with a hearing loss and controls were statistically insignificant (P > 0.05).


Antigen bivalency of antigen-presenting cell-targeted vaccines increases B cell responses.
Antibodies are important for vaccine efficacy. Targeting antigens to antigen-presenting cells (APCs) increases antibody levels. Here, we explore the role of antigen valency in MHC class II (MHCII)-targeted vaccines delivered as DNA. We design heterodimeric proteins that carry either two identical (bivalent vaccines), or two different antigens (monovalent vaccines). Bivalent vaccines with two identical influenza hemagglutinins (HA) elicit higher amounts of anti-HA antibodies in mice than monovalent versions with two different HAs. Bivalent vaccines increase the levels of germinal center (GC) B cells and long-lived plasma cells. Only HA-bivalent vaccines completely protect mice against challenge with homologous influenza virus. Similar results are obtained with other antigens by targeting CD11c and Xcr1 on dendritic cells (DCs) or when administering the vaccine as protein with adjuvant. Bivalency probably increases B cell responses by cross-linking BCRs in readily observable DC-B cell synapses. These results are important for generating potent APC-targeted vaccines.


A Middle-Range Theory for Nurses to Diagnose Ineffective Infant Feeding Patterns.
To construct a middle-range theory, based on the Roy adaptation model, for nurses can diagnose an ineffective infant feeding pattern. Scientific findings were used through a literature review and the concepts of the Roy model. The theory indicated new elements for the diagnosis: 12 etiological factors and 13 clinical indicators. Most of those are not contained in the NANDA taxonomy. The middle-range theory for these diagnoses highlights the need for a review of the components present in the taxonomy. This approach guides research on nursing diagnoses to revise the content of the theory and to expand its concepts and elements for the diagnosis of an ineffective infant feeding pattern.


Single-molecule FRET studies of RNA folding: a Diels-Alderase ribozyme with photolabile nucleotide modifications.
Enzymology at the single-molecule level by using fluorescence resonance energy transfer (smFRET) offers unprecedented insight into mechanistic aspects of catalytic reactions. Implementing spatiotemporal control of the reaction by using an external trigger is highly valuable in these challenging experiments. Here, we have incorporated a light-cleavable caging moiety into specific nucleotides of the Diels-Alderase (DAse) ribozyme. In this way, the folding energy landscape was significantly perturbed, and the catalytic activity was essentially suppressed. A careful smFRET efficiency histogram analysis at various Mg(2+) ion concentrations revealed an additional intermediate state that is not observed for the unmodified DAse ribozyme. We also observed that only a fraction of DAse molecules returns to the native state upon cleavage of the caged group by UV light. These constructs are attractive model RNA systems for further real-time single-molecule observation of the coupling between conformational changes and catalytic activity.


Multiple intracranial hemorrhages at the time of a transiently prolonged activated partial thromboplastin time in an infant with congenital factor VII deficiency.
Factor VII (FVII) deficiency is a rare autosomal recessive hereditary disorder characterized by a normal partial thromboplastin time and a prolonged prothrombin time. For definitive diagnosis, the specific FVII level should be investigated. We report on a 7-month-old boy with congenital FVII deficiency suffering from convulsions and intracerebral hemorrhage. Hematologic tests revealed prolonged prothrombin time associated with a decreased FVII level of 1.7%. Computerized tomography of the brain revealed multifocal hemorrhagic lesions. To our knowledge, multifocal intracranial hemorrhages at the time of a transiently prolonged partial thromboplastin time of unknown origin in a child with congenital FVII deficiency of about 2% has not been reported so far.


First-in-human imaging and kinetic analysis of vesicular acetylcholine transporter density in the heart using [18F]FEOBV PET.
In contrast to cardiac sympathetic activity which can be assessed with established PET tracers, there are currently no suitable radioligands to measure cardiac parasympathetic (cholinergic) activity. A radioligand able to measure cardiac cholinergic activity would be an invaluable clinical and research tool since cholinergic dysfunction has been associated with a wide array of pathologies (e.g., chronic heart failure, myocardial infarction, arrythmias). [18F]Fluoroethoxybenzovesamicol (FEOBV) is a cholinergic radiotracer that has been extensively validated in the brain. Whether FEOBV PET can be used to assess cholinergic activity in the heart is not known. Hence, this study aimed to evaluate the properties of FEOBV for cardiac PET imaging and cholinergic activity mapping. PET data were collected for 40 minutes after injection of 230 ± 50 MBq of FEOBV in four healthy participants (1 female; Age: 37 ± 10; BMI: 25 ± 2). Dynamic LV time activity curves were fitted with Logan graphical, 1-tissue compartment, and 2-tissue compartment models, yielding similar distribution volume estimates for each participant. Our initial data show that FEOBV PET has favorable tracer kinetics for quantification of cholinergic activity and is a promising new method for assessing parasympathetic function in the heart.


The visual electrically evoked potential (VEEP): steady-state responses.
The steady-state response of the visual system may provide us with complementary information about the system, when compared with transient EPs (Regan 1977). With our setup we could record such steady-state responses to electrical stimulation of the visual system for frequencies higher than about 20 Hz. We found comparatively little decline in amplitude with increasing frequency. We conclude that not much filtering is done by the system under study. The absence of pseudoflash related potentials substantiates this conclusion. Therefore it is likely that there is a constant latency for the frequencies under consideration. We also found that after some 10 cycles of the stimulating alternating current the steady-state seemed to be established. This indicates that by introducing silent periods after a limited number of stimulus cycles, recording during these periods and applying the analysis method of Diamond, the latency of the visual electrically evoked steady-state response can be determined.


Delay in gastric emptying rate enhances bioavailability of sodium salicylates in rabbit.
The effect of delay in gastric emptying rate (GER) caused by oil on the bioavailability of sodium salicylate was investigated. The bioavailability of sodium salicylate administered in aqueous, glycerin and oily vehicles was compared in rabbits. Glycerin (having no effect on GER and more viscous than oil) was used to eliminate the effect of viscosity of the oil on the bioavailability of the drug. Glycerin formulation was compared with both aqueous and oily formulations using a two-way crossover test in eight rabbits, in each study. The results indicate that the oily formulation gave a lower rate of absorption and a greater extent of absorption than the other formulations. No significant difference in the rate and extent of absorption was shown between the two vehicles having no effect on GER (aqueous and glycerin vehicles). In conclusion, Delaying effect of the oil on the GER enhances significantly the bioavailability of salicylate. This enhancement is due to the physiological effect of the oil on the GER and not due to its viscosity. Possible reasons for the differences are discussed with particular reference to the effects of oil on the gastric emptying rate (GER).


Dendritic cells in tolerance induction for the treatment of autoimmune diseases.
Autoimmune diseases are characterized by the loss of tolerance toward self-antigens and the induction of destructive immune responses leading to tissue damage. Most patients with autoimmune diseases are treated with immunosuppressive drugs that suppress the immune response in a non-specific fashion, which is inevitably accompanied by several side effects. Antigen-specific immunomodulation and patient-tailored therapies are likely to solve these issues and to elicit long-term protection against disease flares. This Viewpoint analyzes the potential use of DC for induction of antigen-specific tolerance in autoimmune disease settings.


Treatment-resistant depression prior to the diagnosis of cryptococcal meningitis: a case report.
Comorbidity of chronic infectious disorders is one of the common causes of treatment-resistant depression. Depression may alter some aspects of immunity that can contribute to the development of infection. Here we describe an elderly male with treatment-resistant depression. Ten months after antidepressants were administered, he was found to have cryptococcal meningitis. After successful treatment of the central nervous system infection, his depressive symptoms improved apparently. A possible interaction between depression and cellular immunity was discussed. Physicians should be cautious about the risk of opportunistic infection in patients with depression, especially in immunocompromised condition.