A resource for transcriptomic analysis in the mouse brain.
The transcriptome of the cerebral cortex is remarkably homogeneous, with variations being stronger between individuals than between areas. It is thought that due to the presence of many distinct cell types, differences within one cell population will be averaged with the noise from others. Studies of sorted cells expressing the same transgene have shown that cell populations can be distinguished according to their transcriptional profile. We have prepared a low-redundancy set of 16,209 full-length cDNA clones which represents the transcriptome of the mouse visual cortex in its coding and non-coding aspects. Using an independent tag-based approach, CAGE, we confirmed the cortical expression of 72% of the clones. Clones were amplified by PCR and spotted on glass slides, and we interrogated the microarrays with RNA from flow-sorted fluorescent cells from the cerebral cortex of parvalbumin-egfp transgenic mice. We provide an annotated cDNA clone collection which is particularly suitable for transcriptomic analysis in the mouse brain. Spotting it on microarrays, we compared the transcriptome of EGFP positive and negative cells in a parvalbumin-egfp transgenic background and showed that more than 30% of clones are differentially expressed. Our clone collection will be a useful resource for the study of the transcriptome of single cell types in the cerebral cortex.


Distribution of the phospholipase A2 receptor messenger RNA in human gestational tissues.
Recently, a model has been proposed for the involvement of secretory phospholipase A2 (sPLA2) in the onset of and/or progression of human labour via the metabolism of cell membrane glycerophospholipids to generate biologically active, phospholipid-derived mediators. The recent molecular cloning and characterization of a cell-surface receptor for sPLA2 raise the possibility that sPLA2 enzymes may also affect cell function in intrauterine tissues via a receptor-mediated pathway. The aim of this study was to determine the expression profile of the PLA2 receptor messenger RNA in human gestational tissues at term. Messenger RNA for the PLA2 receptor was detected in amnion, choriodecidua and placenta by RT-PCR and transcripts of similar size to the 6.5- and 5.4-kb transcripts previously reported in various other human tissues were detected in choriodecidua by Northern blot analysis. However, smaller transcripts of approximately 4, 2.3 and 1 kb were also detected in choriodecidua by Northern blot analysis and the 2.3-kb transcript and the 1-kb transcript were the only major transcripts detected in amnion and placenta, respectively. The presence of PLA2 receptor mRNA in human gestational tissues indicates that an alternative non-catalytic pathway may contribute to the regulatory effects of sPLA2 isozymes in these tissues. While the specificity and affinity of the various transcripts identified in this study have yet to be determined, PLA2 isozymes released from human gestational tissues during pregnancy and at the time of labour may function as paracrine or autocrine mediators to affect cell function.


Ocular Surface Expression of SARS-CoV-2 Receptors.
The spike proteins of SARS-CoV-2 interact with ACE2 or basigin/CD147 receptors, regulating human-to-human transmissions of COVID-19 together with serine protease TMPRSS2. The expression of these receptors on the ocular surface is unknown. Gene expression of SARS-CoV-2 receptors was investigated in conjunctival epithelial cell samples and in ex-vivo cornea samples using microarray or transcriptome sequencing. ACE2 is expressed in conjunctival samples at a low level, while BSG and TMPRSS2 are expressed at intermediate levels in both conjunctiva and cornea. Other receptors such as ANPEP, AGTR2 are expressed at low level in the conjunctiva. Two RNA editing enzymes involved in antiviral responses, APOBEC3A, and ADAR-1 were also highly expressed. The ocular surface may represent an entry point for the SARS-CoV-2 in the human body. The conjunctiva and the cornea can adopt antiviral countermeasures which may explain the low prevalence of eye involvement.


Beyond heparin and warfarin: the new generation of anticoagulants.
Heparin and warfarin are widely used for the prevention and treatment of venous and arterial thromboembolism. Although effective, both agents have important limitations; for example, both drugs must be monitored, which is inconvenient for patients and for physicians. Heparin requires parenteral administration and can cause heparin-induced thrombocytopenia, an immune-mediated process that can lead to life-threatening thrombosis. Warfarin also has its limitations. Due to its slow onset of action, warfarin must be overlapped with heparin (or another rapidly acting anticoagulant) when treating patients with established thrombosis or who are at high risk for thrombosis. Warfarin dosing is variable because its activity is influenced by dietary intake of vitamin K, genetic polymorphisms in enzymes that are involved in its metabolism and numerous drug-drug interactions that promote or reduce its activity. New anticoagulants have been developed to overcome these problems. Building on a better understanding of coagulation pathways, advances in structure-based drug design and information derived from natural anticoagulants isolated from hematophagous organisms, most of the new anticoagulants target specific coagulation enzymes. Focussing on drugs that have at least completed Phase II evaluation, this article briefly reviews the coagulation pathways and its natural regulators; outlines the limitations of existing anticoagulants and identifies the opportunities for new ones; highlights the properties of selected new anticoagulants; describes the clinical trial results with these agents; and provides a perspective on their potential strengths and weaknesses.


Placebo effect and athletes.
Anecdotal and experimental evidence suggest that the placebo effect is seen in athletics. To this end, placebos have been used in clinical practice and athletic performance. Positive reinforcement and the beliefs of athletes might contribute to the magnitude and direction of the effect, while supplement color, cost, and method of delivery also might be salient factors. Physiological changes similar to those resulting from active medication have been observed as the result of the administration of placebos, raising questions about placebo mechanisms. Researchers should look at design methods that will isolate placebo effects from those of experimental medication.


Adaptation to dynamic faces produces face identity aftereffects.
Face aftereffects are well established for static stimuli and have been used extensively as a tool for understanding the neural mechanisms underlying face recognition. It has also been argued that adaptive coding, as demonstrated by face aftereffects, plays a functional role in face recognition by calibrating our face norms to reflect current experience. If aftereffects tap high-level perceptual mechanisms that are critically involved in everyday face recognition then they should also occur for moving faces. Here we asked whether face identity aftereffects can be induced using dynamic adaptors. The face identity aftereffect occurs when adaptation to a particular identity (e.g., Dan) biases subsequent perception toward the opposite identity (e.g., antiDan). We adapted participants to video of real faces that displayed either rigid, non-rigid, or no motion and tested for aftereffects in static antifaces. Adapt and test stimuli differed in size, to minimize low-level adaptation. Aftereffects were found in all conditions, suggesting that face identity aftereffects tap high-level mechanisms important for face recognition. Aftereffects were not significantly reduced in the motion conditions relative to the static condition. Overall, our results support the view that face aftereffects reflect adaptation of high-level mechanisms important for real-world face recognition in which faces are moving.


Evidence of ectokinase-mediated phosphorylation of osteopontin and bone sialoprotein by osteoblasts during bone formation in vitro.
Osteopontin (OPN) and bone sialoprotein (BSP) are phosphorylated glycoproteins that, together with osteonectin/secreted protein, acidic, rich in cysteine (SPARC) and osteocalcin, comprise the major non-collagen proteins of bone. Although phosphorylation of OPN and BSP, which is known to influence the biological properties of these proteins, has been shown to occur intracellularly, recent studies have demonstrated ectokinase activity in bone cell populations [Mikuni-Takagaki, Kakai, Satoyoshi, Kawano, Suzuki, Kawase and Saito (1995) J. Bone Miner. Res. 10, 231-241]. To determine whether OPN and BSP are phosphorylated by ectokinase activity we have used [gamma-32P]ATP and [gamma-32P]GTP as cell-impenetrable phosphate donors to analyse for ectokinase activity in osteoblastic UMR106.06 cells and fetal rat calvarial cells (FRCCs). By pulse-labelling confluent cells with radiolabelled nucleotides, the phosphorylation of endogenous and exogenously added OPN and BSP was demonstrated together with the labelling of a number of cell surface proteins. These phosphorylation reactions were inhibited by a cell-impermeable ectokinase inhibitor, K252b, and cell surface phosphorylation was also inhibited by exogenously added OPN and BSP substrates, indicating competition for the ectokinase enzyme. However, phosphorylation of OPN and BSP, both of which can mediate cell attachment through Arg-Gly-Asp (RGD) motifs, was not inhibited by an RGD peptide, suggesting that binding of OPN and BSP to cell surface integrins is not required. In similar experiments, ectokinase-mediated phosphorylation of OPN and BSP was demonstrated during mineralized tissue formation by FRCCs in vitro. These studies demonstrate that OPN and BSP secreted by bone cells are phosphorylated by a casein kinase II-like ectokinase present on the surface of osteoblastic cells.


Maintaining and repeating tocolysis: A reflection on evidence.
It is inherent to human logic that both doctors and patients want to suppress uterine contractions when a woman presents in threatened preterm labor. Tocolysis is widely applied in women with threatened preterm labor with a variety of drugs. According to literature, tocolysis is indicated to enable transfer to a tertiary center as well as to ensure the administration of corticosteroids for fetal maturation. There is international discrepancy in the content and the implementation of guidelines on preterm labor. Tocolysis is often maintained or repeated. Nevertheless, the benefit of prolonging pregnancy has not yet been proven, and it is not impossible that prolongation of the pregnancy in a potential hostile environment could harm the fetus. Here we reflect on the use of tocolysis, focusing on maintenance and repeated tocolysis, and compare international guidelines and practices to available evidence. Finally, we propose strategies to improve the evaluation and use of tocolytics, with potential implications for future research.


Assessment of Nutritional Problems in Pediatric Patients with Cancer and the Information Needs of Their Parents: A Parental Perspective.
The majority of problems and symptoms occur in the gastrointestinal system in children with cancer. Parents have difficulty in coping with the nutritional problems and changing routines of children and need support in this respect. This study aimed to assess the nutritional problems of children with cancer and the information needs of their parents. This descriptive study was performed among children with cancer aged 3-18 years and their parents (n = 69). The data were collected through a data collection form developed by the researchers based on the literature. The most prominent nutritional problems experienced by children were loss of appetite (85.5%), nausea (84.1%), vomiting (81.2%), fatigue (79.7%), and mucositis (66.7%). According to the parents, the factors causing these nutritional problems in children were physiological factors (100%) and the foods given to children in the hospital (65.2%). The parents mostly needed information about food-drug interactions (58.0%), food-disease interactions (52.2%), foods that children with neutropenia should avoid or should eat (neutropenic diet) (46.4%), and frequency of nutritional intake (36.2%). This study has shown that most children experience at least one nutritional problem, and the parents need comprehensive and regular information about nutrition. Pediatric oncology nurses have a significant responsibility in the evaluation, education, and monitoring of these children.


Aloe Genus Plants: From Farm to Food Applications and Phytopharmacotherapy.
Aloe genus plants, distributed in Old World, are widely known and have been used for centuries as topical and oral therapeutic agents due to their health, beauty, medicinal, and skin care properties. Among the well-investigated Aloe species are A. arborescens, A. barbadensis, A. ferox, and A. vera. Today, they account among the most economically important medicinal plants and are commonly used in primary health treatment, where they play a pivotal role in the treatment of various types of diseases via the modulation of biochemical and molecular pathways, besides being a rich source of valuable phytochemicals. In the present review, we summarized the recent advances in botany, phytochemical composition, ethnobotanical uses, food preservation, and the preclinical and clinical efficacy of Aloe plants. These data will be helpful to provide future directions for the industrial and medicinal use of Aloe plants.


Thermostable Lactonases Inhibit Pseudomonas aeruginosa Biofilm: Effect In Vitro and in Drosophila melanogaster Model of Chronic Infection.
Pseudomonas aeruginosa is one of the six antimicrobial-resistant pathogens known as "ESKAPE" that represent a global threat to human health and are considered priority targets for the development of novel antimicrobials and alternative therapeutics. The virulence of P. aeruginosa is regulated by a four-chemicals communication system termed quorum sensing (QS), and one main class of QS signals is termed acylhomoserine lactones (acyl-HSLs), which includes 3-Oxo-dodecanoil homoserine lactone (3-Oxo-C12-HSL), which regulates the expression of genes implicated in virulence and biofilm formation. Lactonases, like Paraoxonase 2 (PON2) from humans and the phosphotriesterase-like lactonases (PLLs) from thermostable microorganisms, are able to hydrolyze acyl-HSLs. In this work, we explored in vitro and in an animal model the effect of some lactonases on the production of Pseudomonas virulence factors. This study presents a model of chronic infection in which bacteria were administered by feeding, and Drosophila adults were treated with enzymes and the antibiotic tobramycin, alone or in combination. In vitro, we observed significant effects of lactonases on biofilm formation as well as effects on bacterial motility and the expression of virulence factors. The treatment in vivo by feeding with the lactonase SacPox allowed us to significantly increase the biocidal effect of tobramycin in chronic infection.


Helicobacter pylori infection in children.
Helicobacter pylori infection affects more than half of the world population and it occurs generally in childhood. It is associated with gastroduodenal ulcer, gastric atrophy, intestinal metaplasia, gastric adenocarcinoma and lymphoid tissue-associated lymphoma. It is difficult to eradicate this bacterium due to its high antimicrobial resistance. In children, the infection is asymptomatic in the majority of cases and complications are less common. Probable inverse relationships with allergic diseases and inflammatory bowel diseases are being studied. These reasons mean that the decision to diagnose and treat the infection in children is only considered in specific circumstances in which it provides true benefits. This review focuses on some current considerations regarding epidemiology, diagnosis and treatment of childhood infection, emphasising outcomes and treatment schemes in children.


Self-Management Support in Chronic Care: Practice Implementation Lessons for Healthcare Providers from an Atlantic Collaborative.
In Atlantic Canada, people live with greater risk factors and higher rates of chronic disease than the average Canadian; and health system costs have historically risen faster than other parts of the country. Many clinicians endorse self-management support (SMS) as a means to help patients manage their chronic conditions but often lack the confidence and proper expertise to do so due to limited literature on SMS implementation. This paper draws on two case examples from Atlantic Canada to address gaps between effective SMS interventions and the implementation and evaluation of such interventions that can support provider adoption.


Modeling Dominant and Recessive Forms of Retinitis Pigmentosa by Editing Three Rhodopsin-Encoding Genes in Xenopus Laevis Using Crispr/Cas9.
The utility of Xenopus laevis, a common research subject for developmental biology, retinal physiology, cell biology, and other investigations, has been limited by lack of a robust gene knockout or knock-down technology. Here we describe manipulation of the X. laevis genome using CRISPR/Cas9 to model the human disorder retinitis pigmentosa, and to introduce point mutations or exogenous DNA sequences. We introduced and characterized in-frame and out-of-frame insertions and deletions in three genes encoding rhodopsin by co-injection of Cas9 mRNA, eGFP mRNA, and single guide RNAs into fertilized eggs. Deletions were characterized by direct sequencing and cloning; phenotypes were assessed by assays of rod opsin in retinal extracts, and confocal microscopy of cryosectioned and immunolabeled contralateral eyes. We obtained germline transmission of editing to F1 offspring. In-frame deletions frequently caused dominant retinal degeneration associated with rhodopsin biosynthesis defects, while frameshift phenotypes were consistent with knockout. We inserted eGFP or point mutations into rhodopsin genes by co-injection of repair fragments with homology to the Cas9 target sites. Our techniques can produce high frequency gene editing in X. laevis, permitting analysis in the F0 generation, and advancing the utility of X. laevis as a subject for biological research and disease modeling.


Age-Related Changes in Femoral Head Trabecular Microarchitecture.
Osteoporosis is a prevalent bone condition, characterised by low bone mineral density and increased fracture risk. Currently, the gold standard for identifying osteoporosis and increased fracture risk is through quantification of bone mineral density using dual energy X-ray absorption. However, many studies have shown that bone strength, and consequently the probability of fracture, is a combination of both bone mass and bone 'quality' (architecture and material chemistry). Although the microarchitecture of both non-fracture and osteoporotic bone has been previously investigated, many of the osteoporotic studies are constrained by factors such as limited sample number, use of ovariectomised animal models, and lack of male and female discrimination. This study reports significant differences in bone quality with respect to the microarchitecture between fractured and non-fractured human femur specimens. Micro-computed tomography was utilised to investigate the microarchitecture of femoral head trabecular bone from a relatively large cohort of non-fracture and fracture human donors. Various microarchitectural parameters have been determined for both groups, providing an understanding of the differences between fracture and non -fracture material. The microarchitecture of non-fracture and fracture bone tissue is shown to be significantly different for many parameters. Differences between sexes also exist, suggesting differences in remodelling between males and females in the fracture group. The results from this study will, in the future, be applied to develop a fracture model which encompasses bone density, architecture and material chemical properties for both female and male tissues.


Irritable Bowel Syndrome: Clinical Manifestations, Dietary Influences, and Management.
Irritable bowel syndrome (IBS) is a functional gastrointestinal disorder that is characterized by symptoms of chronic abdominal pain and altered bowel habits in the absence of an overtly identifiable cause. It is the most commonly diagnosed functional gastrointestinal disorder, accounting for about one third of gastroenterology visits. It generally presents as a complex of symptoms, including psychological dysfunction. Hypersensitivity to certain foods, especially foods that contain high amounts of fructose, plays a role in the pathophysiology of IBS. Elevated consumption of high-fructose corn syrup (HFCS) has been discussed in this aspect. The treatment options for IBS are challenging and varied. In addition to dietary restrictions for HFCS-induced IBS, such as low-FODMAP (Fermentable Oligosaccharides, Disaccharide, Monosaccharides, and Polyols) diets, existing drug therapies are administered based on the predominant symptoms and IBS-subtype. Patients with IBS are likely to suffer from issues, such as anxiety, depression, and post-traumatic-stress disorder. Biopsychosocial factors particularly socioeconomic status, sex, and race should, thus, be considered for diagnostic evaluation of patients with IBS.


Navigating triage to meet targets for waiting times.
An initial assessment process, called navigation, has been introduced at Medway Maritime Hospital emergency department (ED) to address problems with the triage system and to meet the latest ED quality clinical indicators. This article explains the rationale for introducing the new assessment process, describes the system and discusses the change-management process needed to implement it. -


Increased risk of dental trauma in patients with allergic rhinitis: A nationwide population-based cohort study.
Allergic rhinitis (AR) is associated with various developmental issues that affecting dentition. We aimed to determine whether AR is associated with an increased risk of traumatic dental injuries (TDIs) in Taiwanese individuals. We used the Taiwan National Health Insurance Research Database (NHIRD) to conduct a nested case-control study. We compared an AR cohort with a matched cohort of patients without AR. New TDI cases were determined during our study period. To compare TDI risk between our study cohorts, we used Cox proportional regression analysis, and hazard ratios (HR) with 95% confidence intervals (CI) were calculated to quantify the association between AR exposure and TDI risk. In total, 76749 patients with AR (31715 male; 45034 female) were identified. In the AR and the non-AR cohorts, 312 patients in total had TDI. Patients with AR had a significantly higher risk of TDI than those without AR (aHR = 1.92; 95% CI = 1.459-2.525; P < 0.001). The risk of TDI was markedly higher in the AR cohort, except in the 3-12-year-old group, and with a CCI ≥ 1. AR patients had a future risk of TDI, indicating a potentially linked disease pathophysiology. The association between AR and TDI is greater among general patients. Clinicians and caregivers should be aware of potential TDI co-morbidity in patients with AR.


Paramagnetic effects on solid state carbon-13 nuclear magnetic resonance spectra of soil organic matter.
The effects of paramagnetic species on solid state 13C nuclear magnetic resonance (NMR) spectra were quantified in a series of doping experiments. The degree of signal loss caused by paramagnetic metals was shown to depend not only on the quantity, but also on the nature of the paramagnetic species, as well as the intimacy of contact with the organic substrate and the type of NMR experiment. Two mechanisms of signal loss were distinguished--signal loss via loss of magnetic field homogeneity, which affects all 13C nuclei in a sample, and signal loss via interaction between electronic and nuclear spins, the effects of which were localized to the close environment of the paramagnetic species. Loss of field homogeneity is important for manganese species, but not for copper species, and is equally important for both cross polarization and Bloch decay experiments. The interaction between electronic and nuclear spins is highly dependent on the spin-lattice relaxation rate constant of the free electron (T1e), as cations with very short T1e values (e.g., Pr3+) cause less signal loss than cations with longer T1e values (e.g., Cu2+, Mn2+). Cross polarization spectra are shown to be more susceptible than Bloch decay spectra to this mechanism of signal loss. Signal loss and increased relaxation rates brought about by paramagnetic species can be used to provide information on soil organic matter (SOM) heterogeneity in the submicron range. This is demonstrated for SOM doped with paramagnetic cations where selective signal loss and increased relaxation rates are used to determine the nature of cation exchange sites.


Standard versus limited colon resection for high risk T1 colon cancer. A matched case-control study.
The National Comprehensive Cancer Network (NCCN) recommends a colectomy in presence of high risk T1 colon polyps considering the risk of incomplete lymph node dissection or presence of residual disease. We evaluated the outcomes of segmental versus standard colon resection for high risk T1 colon cancers, in order to demonstrate if segmental colectomy (SegCR) allows same short-term and oncological results compared to standard radical colectomy (StaCR). METHODS. A matched case-control study on patients who had undergone segmental versus standard colon resection was performed. One-hundred and two patients with high risk T1 colon cancer after endoscopic polypectomy, divided in 2 homogeneous groups of 51 cases, were analyzed and intra-operative, post-operative and oncological data were compared. RESULTS. Segmental colectomy allowed less operative time and intra-operative blood loss compared to StaCR (p < 0.001). Hospital stay after SegCR was shorter compared to StaCR (p < 0.001). No differences were found in terms of overall morbidity and mortality rates. Five-year actuarial overall, disease-free and disease-specific survival after StaCR were similar to SegCR (87%, 96% and 95% vs. 88%, 97% and 94%, respectively, p = 0.51, p=0.33, p=0.78). CONCLUSIONS. According to our findings, SegCR can be a valid alternative to StaCR for high risk T1 colon polyps. Segmental colectomy allows better peri-operative outcomes compared to StaCR ensuring the same oncological long-term outcomes.


Family discord, parental depression, and psychopathology in offspring: ten-year follow-up.
To determine the independent effects of parental depression and family discord on psychopathology in offspring at high and low risk for major depression. One hundred eighty-two offspring of depressed or nondepressed parents were followed over 10 years. In direct interviews, parents' and offspring's psychopathology was evaluated by raters blind to parents' clinical status. Five dimensions of family discord-poor marital adjustment, parent-child discord, low family cohesion, affectionless control, and parental divorce-were assessed. Offspring exposed to either parental depression or family discord had higher rates of psychopathology than their counterparts. High-risk offspring had few family discord measures associated with their psychopathology; in low-risk offspring, family discord was associated with all offspring diagnoses. Between the two risk factors, parental depression proved a more important predictor for offspring major depressive disorder (MDD) and anxiety disorder, whereas family discord was a more important predictor for substance use disorder. Parental depression is a strong and consistent risk factor for offspring MDD and anxiety disorder. Without parental depression, offspring have less exposure to family discord and lower rates of psychopathology. In the presence of family discord, rates of MDD, anxiety disorder and substance use disorder increased. When offspring matured into young adulthood, effects of parental depression and family discord persisted.


Lung cancer with epidermal growth factor receptor exon 20 mutations is associated with poor gefitinib treatment response.
Clinical reports about responsiveness to gefitinib treatment in patients of non-small cell lung cancer (NSCLC) with mutations in exon 20 of epidermal growth factor receptor (EGFR) are limited. To increase understanding of the influence of exon 20 mutations on NSCLC treatment with gefitinib, we investigated the clinical features of lung cancer in patients with exon 20 mutations and analyzed the gefitinib treatment response. We surveyed the clinical data and mutational studies of NSCLC patients with EGFR exon 20 mutations in the National Taiwan University Hospital and reviewed the literature reports about EGFR exon 20 mutations and the gefitinib treatment response. Twenty-three patients with mutations in exon 20 were identified. Nine (39%) had coexisting mutations in EGFR exons other than exon 20. Sixteen patients received gefitinib treatment, and a response was noted in 4 patients. The gefitinib response rate of NSCLC with exon 20 mutations was 25%, far lower than those with deletions in exon 19 and L858R mutations. Interestingly, different exon 20 mutations and coexisting mutations seemed to have a different influence on gefitinib response. EGFR exon 20 mutations of NSCLC patients result in poorer responsiveness to gefitinib treatment, but variability exists between different individuals.


[Comparative study of the management of congenital brachymetatarsia with elongation with callotaxis].
Brachymetatarsia is the > 5 mm shortening of the metatarsal parabolic arc of the foot, in one or more metatarsals. It is directly related with the early closure of the epiphyseal cartilage or with early fusion of the epiphyseal line of the distal end of the metatarsal. The fourth metatarsal is usually the most affected one. Females are more commonly affected, with a female to male ratio of 25:1; 72% of cases have bilateral involvement. The causes may be congenital, posttraumatic or result from specific conditions. A prospective, comparative study was conducted of patients with a diagnosis of congenital brachymetatarsia seen in 2007-2008 and 2008-2012. Seven patients were included in the former period: six girls and one boy. Eight patients were included in the latter period: five girls and three boys. The mean shortening achieved in the first group was 21.1 mm; in the second one, 18 mm, without regression. The daily distraction in both groups was 0.5 mm. Metatarsal elongation by means of callotaxis with external fixators is an appropriate procedure for adolescent patients about to achieve epiphyseal closure. The best results are obtained with gradual distraction at a rate of 0.5 mm per day.


Rotational thromboelastometry in patients with acute respiratory distress syndrome owing to coronavirus disease 2019: Is there a viscoelastic fingerprint and a role for predicting thrombosis?
We evaluated rotational thromboelastometry tracings in 44 critically ill coronavirus disease 2019 patients, to determine whether there is a viscoelastic fingerprint and to test the hypothesis that the diagnosis and prediction of venous thromboembolism would be enhanced by the addition of rotational thromboelastometry testing. Rotational thromboelastometry values reflected an increase in clot strength for the EXTEM, INTEM, and FIBTEM assays beyond the reference range. No hyperfibrinolysis was noted. Fibrinolysis shutdown was present but did not correlate with thrombosis; 32% (14/44) of patients experienced a thrombotic episode. For every 1 mm increase of FIBTEM maximum clot formation, the odds of developing thrombosis increased 20% (95% confidence interval, 0-40%, P = .043), whereas for every 1,000 ng/mL increase in D-dimer, the odds of thrombosis increased by 70% (95% confidence interval, 20%-150%, P = .004), after adjustment for age and sex (AUC 0.96, 95% confidence interval, 0.90-1.00). There was a slight but significant improvement in model performance after adding FIBTEM maximum clot formation and EXTEM clot formation time to D-dimer in a multivariable model (P = .04). D-dimer concentrations were more predictive of thrombosis in our patient population than any other parameter. Rotational thromboelastometry confirmed the hypercoagulable state of coronavirus disease 2019 intensive care unit patients. FIBTEM maximum clot formation and EXTEM clot formation time increased the predictability for thrombosis compared with only using D-dimer. Rotational thromboelastometry analysis is most useful in augmenting the information provided by the D-dimer concentration for venous thromboembolism risk assessment when the D-dimer concentration is between 1,625 and 6,900 ng/dL, but the enhancement is modest. Fibrinolysis shutdown did not correlate with thrombosis.


Differential localization of distinct keratin mRNA-species in mouse tongue epithelium by in situ hybridization with specific cDNA probes.
The tongue of the adult mouse is covered by a multilayered squamous epithelium which is continuous on the ventral surface, however interrupted on the dorsal surface by many filiform and few fungiform papillae. The filiform papillae themselves are subdivided into an anterior and posterior unit exhibiting different forms of keratinization. Thus, the entire epithelium shows a pronounced morphological diversity of well recognizable tissue units. We have used a highly sensitive in situ hybridization technique to investigate the differential expression of keratin mRNAs in the tongue epithelium. The hybridization probes used were cDNA restriction fragments complementary to the most specific 3'-regions of any given keratin mRNA. We could show that independent of the morphologically different tongue regions, all basal cells uniformly express the mRNA of a type I 52-kD keratin, typical also for basal cells of the epidermis. Immediately above the homogenous basal layer a vertically oriented specialization of the keratin expression occurs within the morphological tissue units. Thus the dorsal interpapillary and ventral epithelium express the mRNAs of a type II 57-kD and a type I 47-kD keratin pair. In contrast, in the anterior unit of the filiform papillae, only the 47-kD mRNA is present, indicating that this keratin may be coexpressed in tongue epithelium with different type II partners. In suprabasal cells of both, the fungiform papillae and the posterior unit of the filiform papillae, a mRNA of a type I 59-kD keratin could be detected; however, its type II 67-kD epidermal counterpart seems not to be present in these cells. Most surprisingly, in distinct cells of both types of papillae, a type I 50-kD keratin mRNA could be localized which usually is associated with epidermal hyperproliferation. In conclusion, the in situ hybridization technique applied has been proved to be a powerful method for detailed studies of differentiation processes, especially in morphologically complex epithelia.


Dual Function of iPSC-Derived Pericyte-Like Cells in Vascularization and Fibrosis-Related Cardiac Tissue Remodeling In Vitro.
Myocardial interstitial fibrosis (MIF) is characterized by excessive extracellular matrix (ECM) deposition, increased myocardial stiffness, functional weakening, and compensatory cardiomyocyte (CM) hypertrophy. Fibroblasts (Fbs) are considered the principal source of ECM, but the contribution of perivascular cells, including pericytes (PCs), has gained attention, since MIF develops primarily around small vessels. The pathogenesis of MIF is difficult to study in humans because of the pleiotropy of mutually influencing pathomechanisms, unpredictable side effects, and the lack of available patient samples. Human pluripotent stem cells (hPSCs) offer the unique opportunity for the de novo formation of bioartificial cardiac tissue (BCT) using a variety of different cardiovascular cell types to model aspects of MIF pathogenesis in vitro. Here, we have optimized a protocol for the derivation of hPSC-derived PC-like cells (iPSC-PCs) and present a BCT in vitro model of MIF that shows their central influence on interstitial collagen deposition and myocardial tissue stiffening. This model was used to study the interplay of different cell types-i.e., hPSC-derived CMs, endothelial cells (ECs), and iPSC-PCs or primary Fbs, respectively. While iPSC-PCs improved the sarcomere structure and supported vascularization in a PC-like fashion, the functional and histological parameters of BCTs revealed EC- and PC-mediated effects on fibrosis-related cardiac tissue remodeling.


Extracting, quantifying, and comparing dynamical and biomechanical properties of living matter through single particle tracking.
A panoply of new tools for tracking single particles and molecules has led to an explosion of experimental data, leading to novel insights into physical properties of living matter governing cellular development and function, health and disease. In this Perspective, we present tools to investigate the dynamics and mechanics of living systems from the molecular to cellular scale via single-particle techniques. In particular, we focus on methods to measure, interpret, and analyse complex data sets that are associated with forces, materials properties, transport, and emergent organisation phenomena within biological and soft-matter systems. Current approaches, challenges, and existing solutions in the associated fields are outlined in order to support the growing community of researchers at the interface of physics and the life sciences. Each section focuses not only on the general physical principles and the potential for understanding living matter, but also on details of practical data extraction and analysis, discussing limitations, interpretation, and comparison across different experimental realisations and theoretical frameworks. Particularly relevant results are introduced as examples. While this Perspective describes living matter from a physical perspective, highlighting experimental and theoretical physics techniques relevant for such systems, it is also meant to serve as a solid starting point for researchers in the life sciences interested in the implementation of biophysical methods.


Model Initialization in a Tidally Energetic Regime: a Dynamically Adjusted Objective Analysis.
A simple improvement to objective analysis of hydrographic data is proposed to eliminate spatial aliasing effects in tidally energetic regions. The proposed method consists of the evaluation of anomalies from observations with respect to circulation model fields. The procedure is run iteratively to achieve convergence. The method is applied in the Bay of Fundy and compared with traditional objective analysis procedures and dynamically adjusted climatological fields. The hydrographic skill (difference between observed and model temperature and salinity) of the dynamically adjusted objective analysis is significantly improved by reducing bias and correcting the vertical structure. Representation of the observed velocities is also improved. The resulting flow is consistent with the known circulation in the Bay.


Self-assembly of collagen on flat surfaces: the interplay of collagen-collagen and collagen-substrate interactions.
Fibrillar collagens, common tissue scaffolds in live organisms, can also self-assemble in vitro from solution. While previous in vitro studies showed that the pH and the electrolyte concentration in solution largely control the collagen assembly, the physical reasons why such control could be exerted are still elusive. To address this issue and to be able to simulate self-assembly over large spatial and temporal scales, we have developed a microscopic model of collagen with explicit interactions between the units that make up the collagen molecules, as well as between these units and the substrate. We have used this model to investigate assemblies obtained via molecular dynamics deposition of collagen on a substrate at room temperature using an implicit solvent. By comparing the morphologies from our molecular dynamics simulations with those from our atomic-force microscopy experiments, we have found that the assembly is governed by the competition between the collagen-collagen interactions and those between collagen and the substrate. The microscopic model developed here can serve for guiding future experiments that would explore new regions of the parameter space.


Prediction of hypothyroidism after hemithyroidectomy: a biochemical and pathological analysis.
The possibility of postsurgical hypothyroidism after hemithyroidectomy is no longer a new subject. Although many previous studies have mentioned posthemithyroidectomy hypothyroidism, the incidence and contributing factors for hypothyroidism remain uncertain. We intended to evaluate the incidence and the time of development of hypothyroidism after hemithyroidectomy and to analyze the relationship of posthemithyroidectomy hypothyroidism with preoperative biochemical parameters and postoperative histopathological findings. From February 2001 through December 2004, 287 consecutive cases of hemithyroidectomized patients were retrospectively analyzed; 136 of these patients were included in this study after the exclusion criteria were applied. The relationship between posthemithyroidectomy hypothyroidism and several parameters such as sex, age, preoperative free T4, TSH, microsomal antibody (Ab), thyroglobulin antibody (Ab) levels and lymphocytic infiltration of the resected gland was statistically analyzed. Postoperative hypothyroidism developed in 58 patients (42.6%). In hypothyroid group, 11 patients (19%) showed overt hypothyroidism and 47 patients (81%) showed subclinical hypothyroidism. Preoperative TSH value was significantly higher in the hypothyroid group (2.15+/-1.30 microU/ml) compared to the euthyroid group (1.29+/-0.9 microU/ml). Positive ratio of preoperative microsomal Ab and thyroglobulin Ab were significantly higher in hypothyroid group (38.9 and 41.9%) compared to euthyroid group (3.6 and 19.3%) (P <0.05). In addition, patients with a higher grade of lymphocytic infiltration were found to have a higher probability of developing hypothyroidism. About 85% of postoperative hypothyroidism was detected between 1 and 6 months postoperatively. We might predict the possibility of developing the posthemithyroidectomy hypothyroidism especially in case of preoperatively positive microsomal antibody, thyroglobulin antibody and high-grade lymphocytic infiltration of the resected gland. In addition, our findings support the recommendation for regular serum TSH follow-up at least for 12 months after hemithyroidectomy.


X-ray computed tomography for non-invasive dendrochronology reveals a concealed double panelling on a painting from Rubens' studio.
Dating the wood from historical art objects is a crucial step to ascertain their production time, and support or refute attribution to an artist or a workshop. Dendrochronology is commonly used for this purpose but requires access to the tree-ring pattern in the wood, which can be hindered by preparatory layers, polychromy, wax, or integrated frames. Here we implemented non-invasive dendrochronology based on X-ray computed tomography (CT) to examine a painting on panel attributed to Rubens' studio and its presumed dating around 1636 CE. The CT images achieved a resolution of 37.3 micron and revealed a double panelling, which was concealed by oak strips covering all four edges. The back (visible) board is made of deciduous oak (Quercus subg. Quercus), the most common type of wood used in 17th-century Netherlandish workshops, and was dated terminus post quem after 1557 CE. However, the front (original) board used for the painting has been identified through examination of the wood anatomy as a tropical wood, probably Swietenia sp., a species seldom used in Netherlandish paintings, and remains undated. Its very presence attests the global character of 17th-century trade, and demonstrates the use of exotic species in Flemish studios. The date of the oak board refutes previous results and suggests that this board was trimmed to meet the size of the tropical one, having been glued to it for conservation purposes or with deceiving intentions to pretend that the painting was made on an oak panel. These revelations have opened new lines of art historical inquiry and highlight the potential of X-ray CT as a powerful tool for non-invasive study of historical art objects to retrieve their full history.


Detection of iron-porphyrin proteins with a biochemiluminescent method in search of extraterrestrial life.
Iron-porphyrin proteins (catalase, peroxidase, hemoglobin, cytochrome C) represent an important group of redoxenzymes which have vitally important functions in micro-organisms. A biochemiluminescent method was employed for the detection of iron-porphyrin proteins. The reaction of luminol oxidation with H2O2 is accompanied by chemiluminescence. The rate of hydrogen peroxide decomposition increased 10(5)-10(7) -fold in the presence of the above enzymes as compared with ferrous (or ferric) ions. Possible application of this reaction for the detection of iron-porphyrin proteins of microbial origin was studied. Other authors have suggested this reaction for the detection of extraterrestrial life. Kinetics of the above reaction in the presence of iron-porphyrin proteins were shown to differ both in amplitude and duration of the signal from the pattern observed in the presence of non-hemin catalysts. The reaction pattern in the presence of mixed-soil populations is similar to those observed with pure bacterial cultures and individual iron-porphyrin proteins. Photometric tests revealed that among preparations studied the addition of 0.01% lysozyme was the most effective in destroying cell walls in microbial populations. However, removal of cell walls is not a necessary prerequisite for the detection of iron porphyrin since, for effective luminol oxidation with H2O2 the medium should be kept at pH 12.0. Pretreatment of microbial suspensions with ultrasound increased 2-fold the total signal due to iron porphyrins. The above method gives a reproducible signal indicating the presence of iron porphyrins when sterile nutrient media were innoculated with desert soil samples (Repeteck, Kara-Kum) and incubated for 13 hr. The device was able to detect the presence of no less than 10(5) - 10(6) cells per ml. The addition of limonite (Fe2O3 X nH2O) does not result in the appearance of an appreciable signal in the luminol + H2O2 system.


Effect of subthalamic nucleus or globus pallidus interna stimulation on oculomotor function in patients with Parkinson's disease.
Deep brain stimulation (DBS) of either the globus pallidus interna (GPi) or subthalamic nucleus (STN) is similarly effective for treating somatomotor manifestations of Parkinson's disease (PD), but differences in how stimulation of each target affects oculomotor function are poorly understood. We sought to determine if stimulation of the STN, but not the GPi, affects oculomotor function in PD patients. Nineteen PD patients with DBS implants (8 bilateral GPi, 9 bilateral STN and 2 unilateral STN) were studied. Testing was performed with stimulation on, then off. Somatomotor function was tested using the Unified Parkinson's Disease Rating Scale (UPDRS) motor exam. For oculomotor testing, patients performed pro- and antisaccade tasks while monitored with an infrared eye tracker. Saccadic latency, saccadic intrusions, and square-wave jerks (SWJs) were measured for each trial. As expected, UPDRS motor scores improved with both GPi and STN stimulation. With GPi stimulation, there was no significant difference in oculomotor function with stimulation on or off. However, with STN stimulation on, there was a significant increase in the mean number of SWJs/s, as well as a significant decrease in latency for both pro- and antisaccade tasks. Stimulation of either GPi or STN had similar effects on somatomotor function, but only STN stimulation significantly altered oculomotor function.


Recovery from slow inactivation in K+ channels is controlled by water molecules.
Application of a specific stimulus opens the intracellular gate of a K(+) channel (activation), yielding a transient period of ion conduction until the selectivity filter spontaneously undergoes a conformational change towards a non-conductive state (inactivation). Removal of the stimulus closes the gate and allows the selectivity filter to interconvert back to its conductive conformation (recovery). Given that the structural differences between the conductive and inactivated filter are very small, it is unclear why the recovery process can take up to several seconds. The bacterial K(+) channel KcsA from Streptomyces lividans can be used to help elucidate questions about channel inactivation and recovery at the atomic level. Although KcsA contains only a pore domain, without voltage-sensing machinery, it has the structural elements necessary for ion conduction, activation and inactivation. Here we reveal, by means of a series of long molecular dynamics simulations, how the selectivity filter is sterically locked in the inactive conformation by buried water molecules bound behind the selectivity filter. Potential of mean force calculations show how the recovery process is affected by the buried water molecules and the rebinding of an external K(+) ion. A kinetic model deduced from the simulations shows how releasing the buried water molecules can stretch the timescale of recovery to seconds. This leads to the prediction that reducing the occupancy of the buried water molecules by imposing a high osmotic stress should accelerate the rate of recovery, which was verified experimentally by measuring the recovery rate in the presence of a 2-molar sucrose concentration.


Alpha-synuclein, alcohol use disorders, and Parkinson disease: a case-control study.
Collaborative pooled analyses demonstrated that allele length variability of the dinucleotide repeat sequence within the alpha-synuclein gene promoter (SNCA REP1) is associated with Parkinson disease (PD) worldwide. Other studies demonstrated that variability in the SNCA promoter is also associated with alcohol use disorders, but not consistently. Yet other studies demonstrated that alcohol use disorders are inversely associated with PD, but not consistently. The aim of this study was to clarify the patterns of association between REP1 genotype, alcohol use disorders, and PD. Cases were recruited from the Department of Neurology of the Mayo Clinic in Rochester, MN. The controls included unaffected siblings and unrelated controls. We assessed alcohol use via a structured telephone interview and screened for alcohol use disorders using the CAGE questionnaire. REP1 genotyping was performed using an ABI 3730XL platform. Odds ratios (ORs) and 95% confidence intervals (CIs) were determined using conditional logistic regression models. We recruited 893 case-control pairs. There was an increasing risk of PD with increasing SNCA REP1 allele length (OR 1.18 for each REP1 genotype score unit, 95% CI 1.02-1.35; p=0.02). There was a decreasing risk of PD with increasing CAGE score (p=0.01). The association of REP1 score with PD remained significant after adjusting for CAGE score, and the association of CAGE score with PD remained significant after adjusting for REP1 score. There were no pairwise interactions. Our findings suggest that SNCA REP1 genotype and alcohol use disorders are independently associated with PD.


Microarray with micro- and nano-topographies enables identification of the optimal topography for directing the differentiation of primary murine neural progenitor cells.
During development and tissue repair, progenitor cells are guided by both biochemical and biophysical cues of their microenvironment, including topographical signals. The topographical cues have been shown to play an important role in controlling the fate of cells. Systematic investigation of topographical structures with different geometries and sizes under the identical experimental conditions on the same chip will enhance the understanding of the role of shape and size in cell-topography interactions. A simple customizable multi-architecture chip (MARC) array is therefore developed to incorporate, on a single chip, distinct topographies of various architectural complexities, including both isotropic and anisotropic features, in nano- to micrometer dimensions, with different aspect ratios and hierarchical structures. Polydimethylsiloxane (PDMS) replicas of MARC are used to investigate the influence of different geometries and sizes in neural differentiation of primary murine neural progenitor cells (mNPCs). Anisotropic gratings (2 μm gratings, 250 nm gratings) and isotropic 1 μm pillars significantly promote differentiation of mNPCs into neurons, as indicated by expression of β-III-tubulin (59%, 58%, and 58%, respectively, compared to 30% on the control). In contrast, glial differentiation is enhanced on isotropic 2 μm holes and 1 μm pillars. These results illustrate that anisotropic topographies enhance neuronal differentiation while isotropic topographies enhance glial differentiation on the same chip under the same conditions. MARC enables simultaneous cost-effective investigation of multiple topographies, allowing efficient optimization of topographical and biochemical cues to modulate cell differentiation.


3D Reconstruction of Space Objects from Multi-Views by a Visible Sensor.
In this paper, a novel 3D reconstruction framework is proposed to recover the 3D structural model of a space object from its multi-view images captured by a visible sensor. Given an image sequence, this framework first estimates the relative camera poses and recovers the depths of the surface points by the structure from motion (SFM) method, then the patch-based multi-view stereo (PMVS) algorithm is utilized to generate a dense 3D point cloud. To resolve the wrong matches arising from the symmetric structure and repeated textures of space objects, a new strategy is introduced, in which images are added to SFM in imaging order. Meanwhile, a refining process exploiting the structural prior knowledge that most sub-components of artificial space objects are composed of basic geometric shapes is proposed and applied to the recovered point cloud. The proposed reconstruction framework is tested on both simulated image datasets and real image datasets. Experimental results illustrate that the recovered point cloud models of space objects are accurate and have a complete coverage of the surface. Moreover, outliers and points with severe noise are effectively filtered out by the refinement, resulting in an distinct improvement of the structure and visualization of the recovered points.


The contribution of bullying victimisation to the burden of anxiety and depressive disorders in Australia.
There is now a strong body of literature showing that bullying victimisation during childhood and adolescence precedes the later development of anxiety and depressive disorders. This study aimed to quantify the burden of anxiety and depressive disorders attributable to experiences of bullying victimisation for the Australian population. This study updated a previous systematic review summarising the longitudinal association between bullying victimisation and anxiety and depressive disorders. Estimates from eligible studies published from inception until 18 August 2018 were included and meta-analyses were based on quality-effects models. Pooled relative risks were combined with a contemporary prevalence estimate for bullying victimisation for Australia in order to calculate population attributable fractions (PAFs) for the two mental disorder outcomes. PAFs were then applied to estimates of the burden of anxiety and depressive disorders in Australia expressed as disability-adjusted life years (DALYs). The findings from this study suggest 7.8% of the burden of anxiety disorders and 10.8% of the burden of depressive disorders are attributable to bullying victimisation in Australia. An estimated 30 656 DALYs or 0.52% (95% uncertainty interval 0.33-0.72%) of all DALYs in both sexes and all ages in Australia were attributable to experiences of bullying victimisation in childhood or adolescence. There is convincing evidence to demonstrate a causal relationship between bullying victimisation and mental disorders. This study showed that bullying victimisation contributes a significant proportion of the burden of anxiety and depressive disorders. The investment and implementation of evidence-based intervention programmes that reduce bullying victimisation in schools could reduce the burden of disease arising from common mental disorders and improve the health of Australians.


Changes in quality of life among Norwegian school children: a six-month follow-up study.
A considerable gap exists in regard to longitudinal research on quality of life (QoL) in community populations of children and adolescents. Changes and stability of QoL have been poorly examined, despite the fact that children and adolescents undergo profound developmental changes. The aims of the study were to investigate short-term changes in student QoL with regard to sex and age in a school-based sample. A representative Norwegian sample of 1,821 school children, aged 8-16 years and their parents were tested at baseline and 6 months later, using the Inventory of Life Quality for Children and Adolescents (ILC) and the Kinder Lebensqualität Fragebogen (KINDL). Student response rate at baseline was 71.2% and attrition over the follow-up period was 4.6%, and 1,336 parents (70%) completed the follow-up. Change scores between baseline and follow-up evaluations were analysed by means of ANCOVA in regard to sex and age effects. Students in the 8th grade reported a decrease in QoL over the six-month follow-up period as compared to those in the 6th grade with regard to Family and School domains and total QoL on the KINDL. For emotional well-being a significant linear decrease in QoL across grades 6th to 10th was observed. However, student ratings on the Friends and Self-esteem domains did not change significantly by age. Girls reported a higher decrease in their QoL across all grades over the follow-up period than did boys in respect of Self-esteem on the KINDL, and an age-related decrease in total QoL between 6th and 8th grade on the ILC. Parent reports of changes in child QoL were nonsignificant on most of the domains. The observed age and sex-related changes in school children's QoL across the six-month follow-up period should be considered in epidemiological as well as clinical research.


Analyses of toxic metals and essential minerals in the hair of Arizona children with autism and associated conditions, and their mothers.
The objective of this study was to assess the levels of 39 toxic metals and essential minerals in hair samples of children with autism spectrum disorders and their mothers compared to controls. Inductively coupled plasma-mass spectrometry was used to analyze the elemental content of the hair of children with autism spectrum disorders (n=51), a subset of their mothers (n=29), neurotypical children (n=40), and a subset of their mothers (n=25). All participants were recruited from Arizona. Iodine levels were 45% lower in the children with autism (p=0.005). Autistic children with pica had a 38% lower level of chromium (p=0.002). Autistic children with low muscle tone had very low levels of potassium (-66%, p=0.01) and high zinc (31%, p=0.01). The mothers of young children with autism had especially low levels of lithium (56% lower, p=0.005), and the young children (ages 3-6 yr) with autism also had low lithium (-30%, p=0.04). Low iodine levels are consistent with previous reports of abnormal thyroid function, which likely affected development of speech and cognitive skills. Low lithium in the mothers likely caused low levels of lithium in the young children, which could have affected their neurological and immunological development. Further investigations of iodine, lithium, and other elements are warranted.


Health disparities attributable to air pollutant exposure in North Carolina: Influence of residential environmental and social factors.
Understanding the environmental justice implications of the mortality impacts of air pollution exposure is a public health priority, as some subpopulations may face a disproportionate health burden. We examined which residential environmental and social factors may affect disparities in the air pollution-mortality relationship in North Carolina, US, using a time-stratified case-crossover design. Results indicate that air pollution poses a higher mortality risk for some persons (e.g., elderly) than others. Our findings have implications for environmental justice regarding protection of those who suffer the most from exposure to air pollution and policies to protect their health.


N-3 long-chain polyunsaturated fatty acids for optimal function during brain development and ageing.
We and others have demonstrated that provision of n-3 long chain polyunsaturated fatty acids (n-3 LCPs) in preterm and term babies is associated with retinal electrical responses to light stimuli, and to brain cortex related visual acuity maturation, that are similar to those observed in human milk fed infants. Our follow up results in young children suggest that neurodevelopment and cognitive abilities are also enhanced by early provision of n-3 LCPs through breast milk or DHA-fortified foods. Breast fed infants also require n-3 LCPs after weaning to achieve optimal visual acuity at 12 months of age. Good quality evidence supporting a role for n-3 LCP consumption to enhance learning and/or behaviour in school-age children is currently lacking. Evidence supporting the potential importance of n-3 LCP consumption for good cognitive health in older age is now beginning to emerge. Recent cross-sectional surveys have reported that higher fatty fish/n-3 LCP consumption and or higher n-3 LCP blood concentrations are associated with reduced risk of impaired cognitive function. Similarly, prospective cohort studies have shown that increased fish consumption and higher n-3 LCPs in blood lipid sub-fractions are associated with decreased risk of dementia in older people. We are presently conducting a large randomised controlled trial in a group of adults aged 70-79 years to assess whether an n-3 LCP supplement will preserve retinal function and prevent age related cognitive decline.


Brassica Nigra: Ethopharmacological Review of a Routinely Used Condiment.
Brassica nigra belonging to the family Brassicaceae (syn - black mustard) comprises of dried seeds and is widely cultivated in Mediterranean region and various other countries like India and Europe. The Plant is neither reported in any official book nor reported systematically for pharmacological effects. We undertook a structured search of all the official books including pharmacopoeias and ayurvedic textbooks. The data was collected, sorted and compiled to bring the chemical constituents and uses of B. nigra. B. nigra contains glycosinolates which undergoes hydrolysis in the presence of myrosinase enzyme yields allyl isothiocyante. The bitter taste and pungent odor is due to the isothiocyanates. B. Nigra shows various activities that are medically important such as anticancer, anti-bacterial, antifungal, anti-helminthic, protection against renal and hepatic toxicity and can also be used in diabetes treatment. Black mustards are also used in cardiovascular and neurological disorders. The present review explores the historical background including the macroscopic and microscopic characteristics, chemical constituents, pharmacological action, various evaluation parameters, formulations and uses of black mustard.


Gingival hyperplasia: Should drug interaction be blamed for?
Gingival overgrowth (GO) is one of the common findings in clinical practice. There could be several causes including drugs associated with the GO. Carbamazepine (CBZ) and amlodipine are the drugs which are infrequently documented as a cause in inducing the gingival hyperplasia. Certain drugs in the body fluid might limit the population of plaque bacteria and alter their metabolism that in turn induce the inflammatory mediators and also activate the genetic and biochemical factors responsible for gingival fibroblast growth. Drug-induced GO is a side effect with a multifactorial etiology that seems to orchestrate the interaction between drugs and fibroblasts in the gingiva. We describe a case of trigeminal neuralgia with hypertension treated with multiple drugs including amlodipine and CBZ. Although amlodipine is known to be infrequently associated with GO, an association of CBZ with GO is even rarer. Causality analysis on the World Health Organization Uppsala Monitoring Centre's scale indicates a probable association with offending drugs.


Reactivity and recovery from different types of work measured by catecholamines and cortisol: a systematic literature overview.
To review occupational health, laboratory, and sports literature on neuroendocrine reactivity and recovery from mental, combined mental and physical, or physical tasks. A systematic literature search was performed in eight databases. Studies with catecholamines or cortisol as effect variables measured in blood, urine, or saliva were included. After application of inclusion and exclusion criteria, 77 studies from the initial 559 identified were taken into account. In occupational settings it was found that relatively few studies were conclusive about recovery, which formed a contrast with sports research. For reactivity and recovery up to 1 hour after performing the task, half of the studies considered physical tasks and more than two thirds showed incomplete recovery compared with baseline excretion of catecholamines and cortisol. Recovery extending to 3 days after the task was performed was often incomplete for cortisol after combined mentally and physically demanding tasks, and less often after solely mental or physical tasks. This type of recovery was more often incomplete for adrenaline (epinephrine) than for noradrenaline (norepinephrine), which was the case after mental as well as combined mental and physical tasks. The results from laboratory and sports research may be transferable to some occupations, but more research is needed on the course of recovery relative to health effects in occupational settings.


Short communication: Entrapment of Listeria cells within air pockets of ice cream mix matrix may lead to potentially heat-injured cells.
In our previous study, we observed the sporadic presence of potentially heat-injured cells of Listeria innocua in ice cream mix following a selective enrichment protocol. Although injured cells have not yet been reported to cause any disease outbreaks, it is important to understand their presence in heat-treated food matrices. In this study, we propose a possible protective role of air pockets that may help explain the sporadic presence of potentially heat-injured cells following heat treatment. Challenge studies were conducted by inoculating ice cream mix samples (42% total solids, 16.3% fat, 22.2% total sugar, and 3.4% protein) with Listeria innocua (an established surrogate) at a mean spiking level of 4.0 log cfu/g. The inoculated samples were heat-treated at 69°C for 30 min and potentially heat-injured cells were detected using buffered Listeria enrichment broth, followed by plating on modified Oxford and Rapid'LMono agars. Scanning electron microscopy and atomic force microscopy were conducted on the air-dried, spiked ice cream mix samples, before and after the thermal treatment stages. Although direct plating did not reveal any intact cells in the heat-treated ice cream mix, a more sensitive enrichment protocol was able to identify cells that were potentially heat-injured. The scanning electron micrographs showed air pockets of different sizes in the ice cream mix samples. The spiked mix samples before heat treatment showed some Listeria cells unevenly distributed in the mix matrix and some entrapped within the larger air pockets. After heat treatment, scanning electron and atomic force micrographs showed cells entrapped only within the larger air pockets. The mix matrix, however, did not show any Listeria cells. Confirmation of Listeria at all stages of analysis was done using MALDI-TOF. These observations suggest that the Listeria cells could be entrapped within the larger air pockets and thus may undergo inadequate thermal effect. This could have resulted in their detection as potentially heat-injured cells, as evident under the conditions of the experiment. These results are preliminary observations and further studies are necessary to draw conclusions and understand the true implications of these findings.


Point mutations in the paramyxovirus F protein that enhance fusion activity shift the mechanism of complement-mediated virus neutralization.
Parainfluenza virus 5 (PIV5) activates and is neutralized by the alternative pathway (AP) in normal human serum (NHS) but not by heat-inactivated (HI) serum. We have tested the relationship between the fusion activity within the PIV5 F protein, the activation of complement pathways, and subsequent complement-mediated virus neutralization. Recombinant PIV5 viruses with enhanced fusion activity were generated by introducing point mutations in the F fusogenic peptide (G3A) or at a distal site near the F transmembrane domain (S443P). In contrast to wild-type (WT) PIV5, the mutant G3A and S443P viruses were neutralized by both NHS and HI serum. Unlike WT PIV5, hyperfusogenic G3A and S443P viruses were potent C4 activators, C4 was deposited on NHS-treated mutant virions, and the mutants were neutralized by factor B-depleted serum but not by C4-depleted serum. Antibodies purified from HI human serum were sufficient to neutralize both G3A and S443P viruses in vitro but were ineffective against WT PIV5. Electron microscopy data showed greater deposition of purified human antibodies on G3A and S443P virions than on WT PIV5 particles. These data indicate that single amino acid changes that enhance the fusion activity of the PIV5 F protein shift the mechanism of complement activation in the context of viral particles or on the surface of virus-infected cells, due to enhanced binding of antibodies. We present general models for the relationship between enhanced fusion activity in the paramyxovirus F protein and increased susceptibility to antibody-mediated neutralization.


Metabolic Alterations and WNT Signaling Impact Immune Response in HGSOC.
Our study used transcriptomic and metabolomic strategies to determine the molecular profiles of HGSOC patient samples derived from primary tumor and ascites cells. These data identified clinically relevant heterogeneity among and within patients and highlighted global and patient-specific cellular responses to neoadjuvant chemotherapy (NACT). Tissue from 61 treatment-naïve patients with HGSOC were collected. In addition, 11 benign, 32 ascites, and 18 post-NACT samples (matched to the individual patient's pre-NACT sample) were collected. RNA sequencing (RNA-seq) was performed on all samples collected. Two-dimensional spatial proteomic data was collected for two pairs of pre- and post-NACT. Untargeted metabolomics data using GCxGC-MS was generated for 30 treatment-naive tissues. Consensus clustering, analysis of differential expression, pathway enrichment, and survival analyses were performed. Treatment-naïve HGSOC tissues had distinct transcriptomic and metabolomic profiles. The mesenchymal subtype harbored a metabolomic profile distinct from the other subtypes. Compared with primary tumor tissue, ascites showed significant changes in immune response and signaling pathways. NACT caused significant alterations in gene expression and WNT activity, and this corresponded to altered immune response. Overall, WNT signaling levels were inversely correlated with immune cell infiltration in HGSOC tissues and WNT signaling post-NACT was inversely correlated with progression-free survival. Our study concluded that HGSOC is a heterogenous disease at baseline and growing molecular differences can be observed between primary tumor and ascites cells or within tumors in response to treatment. Our data reveal potential exploratory biomarkers relevant for treatment selection and predicting patient outcomes that warrant further research.


A new supervised method for blood vessel segmentation in retinal images by using gray-level and moment invariants-based features.
This paper presents a new supervised method for blood vessel detection in digital retinal images. This method uses a neural network (NN) scheme for pixel classification and computes a 7-D vector composed of gray-level and moment invariants-based features for pixel representation. The method was evaluated on the publicly available DRIVE and STARE databases, widely used for this purpose, since they contain retinal images where the vascular structure has been precisely marked by experts. Method performance on both sets of test images is better than other existing solutions in literature. The method proves especially accurate for vessel detection in STARE images. Its application to this database (even when the NN was trained on the DRIVE database) outperforms all analyzed segmentation approaches. Its effectiveness and robustness with different image conditions, together with its simplicity and fast implementation, make this blood vessel segmentation proposal suitable for retinal image computer analyses such as automated screening for early diabetic retinopathy detection.


Mortality and morbidity after mitral valve repair: the importance of left ventricular dysfunction.
A retrospective study of 219 consecutive patients who underwent mitral valve repair for mitral regurgitation or mixed mitral valve disease with at least moderate regurgitation was undertaken. The etiology was degenerative in 151 (68.9%) patients, endocarditis in 24 (10.9%), rheumatic in 22 (10.0%), ischemic in 13 (5.9%), congenital in five (2.3%) and cardiomyopathy in two (0.9%). The average age was 64.8 +/- 10.9 years, the average follow up 30.2 +/- 24.1 months. Pre-operatively, 74% were in NYHA functional class III or IV. Left ventricular function was assessed by angiography and echocardiography, and moderate or severe impairment (ejection fraction < or = 40%) was assumed to represent significant left ventricular dysfunction. There were six (2.7%) hospital deaths, four of which were due to left ventricular dysfunction. The seven-year mortality and seven-year combined mortality and morbidity were 14.4% and 37.1% respectively for complications related to left ventricular dysfunction, and 2.5% and 24.2% respectively for unrelated complications. Subgroup analysis showed that five-year mortality and five-year combined mortality and morbidity due to left ventricular dysfunction were significantly worse in patients who had pre-operative left ventricular dysfunction than those who did not: 28.5% vs. 6.1% (p < 0.001) and 52.4% vs. 17.8% (p < 0.001). There was nevertheless a significant incidence of postoperative left ventricular dysfunction in patients with satisfactory preoperative left ventricular function. In this group, five-year mortality and five-year combined mortality and morbidity due to left ventricular dysfunction were higher in patients who were in NYHA class III or IV preoperatively than in those who were not: 11.2% vs. 0% (NS) and 25.9% vs. 0% (p < 0.01) respectively, particularly if they also had early (3-10 days) post-operative left ventricular dysfunction: 20.4% (p < 0.001) and 41.7% (p < 0.001) respectively. Despite preservation of the mitral apparatus, left ventricular dysfunction remains a major cause of mortality and morbidity following mitral valve repair for mitral regurgitation.


Hands-free administration of subjective workload scales: acceptability in a surgical training environment.
Subjective workload measures are usually administered in a visual-manual format, either electronically or by paper and pencil. However, vocal responses to spoken queries may sometimes be preferable, for example when experimental manipulations require continuous manual responding or when participants have certain sensory/motor impairments. In the present study, we evaluated the acceptability of the hands-free administration of two subjective workload questionnaires - the NASA Task Load Index (NASA-TLX) and the Multiple Resources Questionnaire (MRQ) - in a surgical training environment where manual responding is often constrained. Sixty-four undergraduates performed fifteen 90-s trials of laparoscopic training tasks (five replications of 3 tasks - cannulation, ring transfer, and rope manipulation). Half of the participants provided workload ratings using a traditional paper-and-pencil version of the NASA-TLX and MRQ; the remainder used a vocal (hands-free) version of the questionnaires. A follow-up experiment extended the evaluation of the hands-free version to actual medical students in a Minimally Invasive Surgery (MIS) training facility. The NASA-TLX was scored in 2 ways - (1) the traditional procedure using participant-specific weights to combine its 6 subscales, and (2) a simplified procedure - the NASA Raw Task Load Index (NASA-RTLX) - using the unweighted mean of the subscale scores. Comparison of the scores obtained from the hands-free and written administration conditions yielded coefficients of equivalence of r=0.85 (NASA-TLX) and r=0.81 (NASA-RTLX). Equivalence estimates for the individual subscales ranged from r=0.78 ("mental demand") to r=0.31 ("effort"). Both administration formats and scoring methods were equally sensitive to task and repetition effects. For the MRQ, the coefficient of equivalence for the hands-free and written versions was r=0.96 when tested on undergraduates. However, the sensitivity of the hands-free MRQ to task demands (η(partial)(2)=0.138) was substantially less than that for the written version (η(partial)(2)=0.252). This potential shortcoming of the hands-free MRQ did not seem to generalize to medical students who showed robust task effects when using the hands-free MRQ (η(partial)(2)=0.396). A detailed analysis of the MRQ subscales also revealed differences that may be attributable to a "spillover" effect in which participants' judgments about the demands of completing the questionnaires contaminated their judgments about the primary surgical training tasks. Vocal versions of the NASA-TLX are acceptable alternatives to standard written formats when researchers wish to obtain global workload estimates. However, care should be used when interpreting the individual subscales if the object is to make comparisons between studies or conditions that use different administration modalities. For the MRQ, the vocal version was less sensitive to experimental manipulations than its written counterpart; however, when medical students rather than undergraduates used the vocal version, the instrument's sensitivity increased well beyond that obtained with any other combination of administration modality and instrument in this study. Thus, the vocal version of the MRQ may be an acceptable workload assessment technique for selected populations, and it may even be a suitable substitute for the NASA-TLX.


PI3K in T Cell Adhesion and Trafficking.
PI3K signalling is required for activation, differentiation, and trafficking of T cells. PI3Kδ, the dominant PI3K isoform in T cells, has been extensively characterised using PI3Kδ mutant mouse models and PI3K inhibitors. Furthermore, characterisation of patients with Activated PI3K Delta Syndrome (APDS) and mouse models with hyperactive PI3Kδ have shed light on how increased PI3Kδ activity affects T cell functions. An important function of PI3Kδ is that it acts downstream of TCR stimulation to activate the major T cell integrin, LFA-1, which controls transendothelial migration of T cells as well as their interaction with antigen-presenting cells. PI3Kδ also suppresses the cell surface expression of CD62L and CCR7 which controls the migration of T cells across high endothelial venules in the lymph nodes and S1PR1 which controls lymph node egress. Therefore, PI3Kδ can control both entry and exit of T cells from lymph nodes as well as the recruitment to and retention of T cells within inflamed tissues. This review will focus on the regulation of adhesion receptors by PI3Kδ and how this contributes to T cell trafficking and localisation. These findings are relevant for our understanding of how PI3Kδ inhibitors may affect T cell redistribution and function.


Potential risk of induced malaria by blood transfusion in South-eastern Nigeria.
Induced malaria by blood transfusion is a potential health hazard but is often neglected in many malaria endemic areas. Standard parasitological technique was used to determine the prevalence of malaria among blood donors in the South-eastern Nigeria. Of the total 325 blood donors (310 males and 15 females) screened, 133 (40.9%, CI 95%: 35.6-46.2%) were infected with malaria parasite, 78 (58.6%) had 1-10 parasites per 100 thick film fields ('+' or 4-40 parasites per mm3) while 55 (41.4%) had 11-100 parasites per 100 thick film fields ('++' or 41-400 parasites per mm3). P. falciparum was identified in all the infected cases, however 3 (2.3%) persons had mixed infection with P. malariae. Males were more infected (41.3%, CI 95%: 35.8-46.8%) than females (33.3%, CI 95%: 9.4-57.2%). The infection decreased with age with highest prevalence of 48.5% among those aged 20-25 years. The infection significantly varied with age but not with sex (P<0.05). Individuals with blood group B were slightly more infected (42.1%, 95%CI., 19.6-64.6%) than those of groups O (41.0%,CI 95%: 35.3-46.7%) and A (40.0%, CI 95%: 20.8-59.2%) but there was no significant difference (P<0.05). Highest prevalence of infection was recorded in the month of April corresponding to the onset of the wet season. An overhaul of existing blood donation policies in many health facilities in the sub-Saharan Africa to incorporate malaria screening is advocated. Curative antimalarial drugs followed by prophylactic drugs should be given to all recipients of parasitized blood.


Classical complement pathway factor alterations in narcolepsy.
Narcolepsy is a chronic sleep disorder long hypothesised to be an autoimmune disease. Complement-mediated immune mechanisms have not been investigated in detail in narcolepsy. Our aim was to establish the significance of classical pathway activation in narcolepsy. Sera of 42 narcolepsy patients and 26 healthy controls were screened with ELISA to determine the levels of C1q, C3a, C4d and complement component 4 binding protein (C4BP). A home-made ELISA method was developed to detect antibodies to C4BP-alpha (anti-C4BPA). The correlation between complement levels and clinical findings was examined. C1q levels were significantly higher in narcolepsy patients while C4d and C4BP levels were significantly lower compared to healthy controls. C3a levels were comparable among patients and controls. Eleven narcolepsy patients showed serum anti-C4BPA levels. Total rapid eye movements (REM) time, sleep onset latency, REM sleep latency, sleep activity, percentage of wakefulness after sleep onset and Epworth sleepiness scale scores were correlated with levels of different complement factors. Complement-mediated immune mechanisms might partake in narcolepsy pathogenesis. The precise role of autoantibodies on complement level alterations needs to be investigated. Levels of complement factors and degradation products may potentially be utilised as biomarkers to predict the clinical severity of narcolepsy.


Myocardial perfusion imaging with thallium-201 to evaluate patients before and after percutaneous transluminal coronary angioplasty.
Thallium-201 imaging may be used to help determine the distribution and amount of myocardium in jeopardy and the success of revascularization after percutaneous transluminal coronary angioplasty. Single photon emission computed tomography is particularly advantageous because of its ability to differentiate vascular territories and thus evaluate patients with multivessel disease. Myocardial infarction resulting from complications such as distal embolization and side-branch occlusion can be detected. Thallium-201 imaging early after angioplasty may show abnormal results caused by transiently insufficient coronary flow reserve. However, studies performed 6 weeks or more after angioplasty accurately detect early restenosis and may identify those individuals likely to become symptomatic and eventually develop restenosis in the future.


Lineage-Specific Expansion of the Chalcone Synthase Gene Family in Rosids.
Rosids are a monophyletic group that includes approximately 70,000 species in 140 families, and they are found in a variety of habitats and life forms. Many important crops such as fruit trees and legumes are rosids. The evolutionary success of this group may have been influenced by their ability to produce flavonoids, secondary metabolites that are synthetized through a branch of the phenylpropanoid pathway where chalcone synthase is a key enzyme. In this work, we studied the evolution of the chalcone synthase gene family in 12 species belonging to the rosid clade. Our results show that the last common ancestor of the rosid clade possessed six chalcone synthase gene lineages that were differentially retained during the evolutionary history of the group. In fact, of the six gene lineages that were present in the last common ancestor, 7 species retained 2 of them, whereas the other 5 only retained one gene lineage. We also show that one of the gene lineages was disproportionately expanded in species that belonged to the order Fabales (soybean, barrel medic and Lotus japonicas). Based on the available literature, we suggest that this gene lineage possesses stress-related biological functions (e.g., response to UV light, pathogen defense). We propose that the observed expansion of this clade was a result of a selective pressure to increase the amount of enzymes involved in the production of phenylpropanoid pathway-derived secondary metabolites, which is consistent with the hypothesis that suggested that lineage-specific expansions fuel plant adaptation.


TAK1 is a pivotal therapeutic target for tumor progression and bone destruction in myeloma.
Along with the tumor progression, the bone marrow microenvironment is skewed in multiple myeloma (MM), which underlies the unique pathophysiology of MM and confers aggressiveness and drug resistance in MM cells. TGF-β-activated kinase-1 (TAK1) mediates a wide range of intracellular signaling pathways. We demonstrate here that TAK1 is constitutively overexpressed and phosphorylated in MM cells, and that TAK1 inhibition suppresses the activation of NF-κB, p38MAPK, ERK and STAT3 to decrease the expression of critical mediators for MM growth and survival, including PIM2, MYC, Mcl-1, IRF4, and Sp1, along with a substantial reduction in the angiogenic factor VEGF in MM cells. Intriguingly, TAK1 phosphorylation was also induced along with upregulation of vascular cell adhesion molecule-1 (VCAM-1) in bone marrow stromal cells (BMSCs) in cocultures with MM cells, which facilitated MM cell-BMSC adhesion while inducing IL-6 production and receptor activator of nuclear factor κ-Β ligand (RANKL) expression by BMSCs. TAK1 inhibition effectively impaired MM cell adhesion to BMSCs to disrupt the support of MM cell growth and survival by BMSCs. Furthermore, TAK1 inhibition suppressed osteoclastogenesis enhanced by RANKL in cocultures of bone marrow cells with MM cells, and restored osteoblastic differentiation suppressed by MM cells or inhibitory factors for osteoblastogenesis overproduced in MM. Finally, treatment with the TAK1 inhibitor LLZ1640-2 markedly suppressed MM tumor growth and prevented bone destruction and loss in mouse MM models. Therefore, TAK1 inhibition may be a promising therapeutic option targeting not only MM cells but also the skewed bone marrow microenvironment in MM.


Smartphone-Guided Needle Angle Selection During CT-Guided Procedures.
In CT-guided intervention, translation from a planned needle insertion angle to the actual insertion angle is estimated only with the physician's visuospatial abilities. An iPhone app was developed to reduce reliance on operator ability to estimate and reproduce angles. The iPhone app overlays the planned angle on the smartphone's camera display in real-time based on the smartphone's orientation. The needle's angle is selected by visually comparing the actual needle with the guideline in the display. If the smartphone's screen is perpendicular to the planned path, the smartphone shows the Bull's-Eye View mode, in which the angle is selected after the needle's hub overlaps the tip in the camera. In phantom studies, we evaluated the accuracies of the hardware, the Guideline mode, and the Bull's-Eye View mode and showed the app's clinical efficacy. A proof-of-concept clinical case was also performed. The hardware accuracy was 0.37° ± 0.27° (mean ± SD). The mean error and navigation time were 1.0° ± 0.9° and 8.7 ± 2.3 seconds for a senior radiologist with 25 years' experience and 1.5° ± 1.3° and 8.0 ± 1.6 seconds for a junior radiologist with 4 years' experience. The accuracy of the Bull's-Eye View mode was 2.9° ± 1.1°. Combined CT and smart-phone guidance was significantly more accurate than CT-only guidance for the first needle pass (p = 0.046), which led to a smaller final targeting error (mean distance from needle tip to target, 2.5 vs 7.9 mm). Mobile devices can be useful for guiding needle-based interventions. The hardware is low cost and widely available. The method is accurate, effective, and easy to implement.


Immune response to porin in cattle immunized with whole cell, outer membrane, and outer membrane protein antigens of Brucella abortus combined with trehalose dimycolate and muramyl dipeptide adjuvants.
The immune response of cattle to nonliving vaccines derived from Brucella abortus rough strain 45/20 was studied. Vaccines contained trehalose dimycolate and a derivative of muramyl dipeptide. N-acetylmuramyl-L-alpha-aminobutyryl-D-isoglutamine. A factorial experiment was designed to test the effects of type of antigen, quantity of antigen, and quantity of mineral oil on the immune response to porin. Muramyl dipeptide was kept constant at 5 mg per dose, and 1 part of trehalose dimycolate was incorporated for two parts of dry matter. Over a 10-week period, blastogenesis responses to porin were largest in cattle immunized with outer membranes; the highest antibody titers to the porin-lipopolysaccharide complex were achieved by immunization with detergent-extracted outer membrane proteins. There was no advantage in the use of 25, rather than 5, mg of any of the antigens, but antibody responses were improved by increasing the quantity of oil from 0.6 to 1.8 ml per dose. In other animals, blastogenesis and antibody responses were sustained at high levels longer than 3 months after two vaccinations with outer membrane proteins. Intradermal injection of porin evoked inflammatory reactions histologically consistent with delayed-type hypersensitivity. Cross-reactions in cases of delayed-type hypersensitivity occurred with porin derived from a smooth strain of B. abortus but were less extensive than in the blastogenesis test. The magnitude of the delayed-type hypersensitivity and blastogenesis responses induced by vaccination exceeded those observed after natural or experimental infections. No ill effects were observed after vaccination. These findings provide a basis for the use of trehalose dimycolate and muramyl dipeptide adjuvants in evaluating nonviable vaccines for bovine brucellosis.


Tissue-engineered flexible ear-shaped cartilage.
Previous attempts to engineer human ear-shaped constructs mimicked human shape but lacked the flexibility and size of a human ear. Recently, the authors engineered flexible cartilage by incorporating a perichondrium-like layer into the construct. In this study, they used lyophilized swine perichondrium as a pseudoperichondrium, examined its ability to confer flexibility to tissue-engineered cartilage, and used it to engineer flexible cartilage in the shape and size of a human ear. Auricular chondrocytes and perichondrium were isolated from swine. Chondrocytes were mixed with fibrin polymer and gelled to form 5 x 20-mm constructs. Constructs alone (control, n = 6) or constructs sandwiched between two layers of lyophilized swine perichondrium (experimental, n = 6) were implanted into athymic mice. Auricular chondrocytes in fibrin polymer and lyophilized perichondrium were also used to form a tri-layer, ear-shaped construct, which was implanted into an athymic rat and externally stented for 6 weeks (n = 1). At 12 weeks, constructs were analyzed with histology and gross mechanical testing. New cartilaginous tissue was engineered in both the experimental and control groups. In samples laminated with lyophilized swine perichondrium, the intimate integration of the laminate with the neocartilage closely resembled the histoarchitecture of the native swine ear. Experimental constructs had mechanical properties similar to those of the native swine ear, while control constructs fractured with similar testing. The engineered ear could not be fractured with gross mechanical testing, and its size, shape, and flexibility remained stable. This study demonstrates that it is possible to engineer a cartilage construct that resembles the human ear not only in shape but also in size and flexibility. This study also confirms that lamination is a reliable method to confer elastic-like flexibility to an engineered cartilage construct.


Epidemiological implications of the isolations and intratypic serodifferentiation of poliovirus strains in the Netherlands.
From the results of virus isolation and serological studies it can be concluded that poliovirus does not occur endemically in the Netherlands. On the other hand continual importation of poliovirus is demonstrated among children of foreign workers, refugees and adoptive children. A certain part of these imported poliovirus strains appear to be wild type as shown by intratypic serodifferentiation with monospecific antisera. These strains are a continuous danger to non-immunized persons lacking neutralizing antibodies. The intratypic serodifferentiation with monospecific antisera also appears to be applicable for the differentiation of wild poliovirus strains and may therefore be a useful tool in epidemiological studies.


Gene expression changes after exposure to six-mix in a mouse model.
The exposure of Libby MT residents to amphibole-contaminated vermiculite is well known. To explore the gene-environment interactions in the development of asbestos-related diseases (ARD), a mouse model of asbestos exposure using Six-mix (a combination of amphibole fibers gathered from six sites at the Libby vermiculite mine), crocidolite asbestos, or saline as a negative control was used to determine both gene expression responses by using mouse 10,000 oligonucleotide array and to visualize these changes histologically. Mice were sacrificed and whole lungs harvested for histology and microarray analysis six months following exposure via intratracheal instillation. Using an arbitrary cutoff of 1.25-fold change, genes whose RNA expression levels were specifically altered in response to the different amphibole exposures were grouped into categories by a gene ontology analysis program, GoMiner. Our hypothesis was that assessment of asbestos-responsive genes would provide a better understanding of response mechanisms. These experiments have provided new candidates for genes involved in the asbestos response pathways.


Volumetric feature extraction and visualization of tomographic molecular imaging.
Electron tomography is useful for studying large macromolecular complex within their cellular context. The associate problems include crowding and complexity. Data exploration and 3D visualization of complexes require rendering of tomograms as well as extraction of all features of interest. We present algorithms for fully automatic boundary segmentation and skeletonization, and demonstrate their applications in feature extraction and visualization of cell and molecular tomographic imaging. We also introduce an interactive volumetric exploration and visualization tool (Volume Rover), which encapsulates implementations of the above volumetric image processing algorithms, and additionally uses efficient multi-resolution interactive geometry and volume rendering techniques for interactive visualization.


Long-term clinical and biochemical residue after COVID-19 recovery.
The long-term health consequences of coronavirus disease 2019 (COVID-19) are still unclear. The majority of previous trials addressed the post-COVID-19 symptoms through comprehensive medical questionnaires for relatively short periods after recovery. We tried to detect the potential pathological clinical signs and biochemical residue which persist for more than 3 months after the negative real-time polymerase chain reaction (RT-PCR) test of SARS-CoV-2. Among 120 COVID-19 survivors of mean age 38.29 and 55.6% male proportion, systolic blood pressure was significantly elevated ( P =0.001). Erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), D-dimer showed higher values in COVID-19 survivors ( P < 0.001). Alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl trans-peptidase (GGT), and alkaline phosphatase (ALP) were significantly elevated in contrast to serum albumin that was reduced in COVID-19 survivors ( P ≤0.001). Serum lipase, amylase and albuminuria were higher in COVID-19 survivors ( P ≤0.001). Regression analysis (AOR, 95% CI) showed that ESR ( P = 0.014), haemoglobin concentration ( P = 0.039), serum lipase ( P = 0.018), blood urea nitrogen ( P = 0.003), albuminuria ( P = 0.046), 25(OH) vitamin D ( P = 0.002), and serum uric acid ( P = 0.005) were the significant predictors of COVID-19 survivors (94.8% an overall prediction). COVID-19 survivors experienced residual significant clinical and biochemical alterations that necessitate comprehensive medical care and close follow-up for longer periods.