3D Printed Vascular Networks Enhance Viability in High-Volume Perfusion Bioreactor.
There is a significant clinical need for engineered bone graft substitutes that can quickly, effectively, and safely repair large segmental bone defects. One emerging field of interest involves the growth of engineered bone tissue in vitro within bioreactors, the most promising of which are perfusion bioreactors. Using bioreactor systems, tissue engineered bone constructs can be fabricated in vitro. However, these engineered constructs lack inherent vasculature and once implanted, quickly develop a necrotic core, where no nutrient exchange occurs. Here, we utilized COMSOL modeling to predict oxygen diffusion gradients throughout aggregated alginate constructs, which allowed for the computer-aided design of printable vascular networks, compatible with any large tissue engineered construct cultured in a perfusion bioreactor. We investigated the effect of 3D printed macroscale vascular networks with various porosities on the viability of human mesenchymal stem cells in vitro, using both gas-permeable, and non-gas permeable bioreactor growth chamber walls. Through the use of 3D printed vascular structures in conjunction with a tubular perfusion system bioreactor, cell viability was found to increase by as much as 50% in the core of these constructs, with in silico modeling predicting construct viability at steady state.


Tracking exocytosis of a GPI-anchored protein in Aspergillus nidulans.
Secretion of the glycosylphosphatidylinositol-anchored protein (GPI-AP) EglC was investigated in the filamentous fungus Aspergillus nidulans, exploiting a sucrose-inducible promoter to conditionally express the protein in cells blocked at different steps of exocytosis. EglC is delivered to the cell surface in a polarized fashion, but appears to redistribute rapidly toward apico-distal regions. Inactivation of SarASar1 mediating COPII vesicle biogenesis resulted in the accumulation of EglC in the endoplasmic reticulum (ER) but, rather than concentrating in ER-exit-sites, the reporter labeled the ER uniformly. Abnormal posttranslational modifications of EglC were detected in sarAts and sed5ts mutants, suggesting that blocking COPII biogenesis or traffic in the ER/Golgi interface might affect GPI remodeling. EglC delivery to the plasma membrane requires, besides Golgi function, the TRAPPII complex mediating the biogenesis of RAB11 secretory vesicles at the TGN, but is unaffected by the absence of RAB5, the key regulator of early endosome biogenesis/maturation. Thus, unlike the soluble extracellular enzyme inulinase, EglC is directly delivered from the TGN to the plasma membrane without involvement of endosomes. We conclude that in A. nidulans, GPI-APs follow a direct secretory pathway from the ER to the plasma membrane.


Resorbed co-twin as an explanation for discrepant chorionic villus results: non-mosaic 47,XX,+16 in villi (direct and culture) with normal (46,XX) amniotic fluid and neonatal blood.
Non-mosaic trisomy 16 was observed in chorionic villus cytotrophoblasts (direct) as well as cultured mesenchymal core cells derived from the pregnancy of a 38-year-old woman. Chromosome preparations from amniotic fluid and neonatal cultures (cord blood) were 46,XX. Normal fetal growth as determined by serial ultrasound examinations occurred throughout the pregnancy, which resulted in a healthy 2724 g female. Multiple biopsies taken from the umbilical cord, placental cotyledons, and fetal membranes were 46,XX. However, a placental nodule and three of six cultures initiated from membranes (amnion and chorion) showed 46,XX/47,XX,+16 mosaicism. We propose that the trisomy 16 cells arose from residual villi derived from a trisomic co-twin that never developed. This case further demonstrates that normal fetal growth may presage normal outcome irrespective of cytogenetic findings in cytotrophoblasts (direct) and cultured mesenchymal core cells.


Expanding insights of mitochondrial dysfunction in Parkinson's disease.
The quest to disentangle the aetiopathogenesis of Parkinson's disease has been heavily influenced by the genes associated with the disease. The alpha-synuclein-centric theory of protein aggregation with the adjunct of parkin-driven proteasome deregulation has, in recent years, been complemented by the discovery and increasing knowledge of the functions of DJ1, PINK1 and OMI/HTRA2, which are all associated with the mitochondria and have been implicated in cellular protection against oxidative damage. We critically review how these genes fit into and enhance our understanding of the role of mitochondrial dysfunction in Parkinson's disease, and consider how oxidative stress might be a potential unifying factor in the aetiopathogenesis of the disease.


Protective effects of vasoactive intestinal peptide (VIP) in ischemic retinal degeneration.
Vasoactive intestinal peptide (VIP) is a pleiotropic neuropeptide, acting as a neuromodulator and neuroprotective peptide in the CNS after injuries. We have previously described that pituitary adenylate cyclase-activating polypeptide (PACAP), another member of the same peptide family, is retinoprotective in ischemic lesions. The aim of this study was to investigate the protective potential of VIP in bilateral common carotid artery occlusion (BCCAO)-induced ischemic retinal lesion. Two-month-old rats were subjected to BCCAO and treated with intravitreal VIP injection. Their retinas were processed for histology after 2 weeks of survival. We measured the number of the cells/100 μm of the ganglion cell layer and the thickness of each layer such as the outer nuclear, outer plexiform, inner nuclear, and inner plexiform layers as well as that of the whole retina. We found that treatment with 1,000 pmol VIP, but not 100 pmol VIP, had significant protective effects in BCCAO-injured retina, as shown by the morphometric analysis. Comparing the neuroprotective effects of VIP and PACAP in BCCAO-operated retinas, PACAP was more effective, already protective at 100-pmol doses. Similar to other studies, we found that VIP must be given at least in 10 times more concentration than PACAP to achieve a similar degree of neuroprotection in the retina.


The roles of EBV-encoded microRNAs in EBV-associated tumors.
Epstein-Barr virus (EBV) is believed to be a pathogen causing a number of human cancers, but the pathogenic mechanisms remain unclear. An increasing number of studies have indicated that EBV-encoded microRNAs (EBV miRNAs) are expressed in a latency type- and tumor type-dependent manner, playing important roles in the development and progression of EBV-associated tumors. By targeting one or more genes of the virus and the host, EBV miRNAs are responsible for the deregulation of a variety of viral and host cell biological processes, including viral replication, latency maintenance, immune evasion, cell apoptosis and metabolism, and tumor proliferation and metastasis. In addition, some EBV miRNAs can be used as excellent diagnostic, prognostic and treatment efficacy predictive biomarkers for EBV-associated tumors. More importantly, EBV miRNA-targeting therapeutics have emerged and have been developing rapidly, which may open a new era in the treatment of EBV-associated tumors in the near future.


[Change in the selectivity of the sodium channels of a nerve fiber membrane under the action of veratrine].
The ionic currents of the nodal membrane were measured under voltage clamp conditions. The membrane being +40 mv. The replacing of the external Na+-ions to K+- and NH4+-ions have showed that the relative pearmeabilities of the veratrine-modified channels calculated from the constant field theory are arranged in the following row: PNa:PK:PNH4 = 1:0.29:0.61, which differs from the same row for the normal channels. The decreasing of the slope of current-voltage relations of the modified channels with the replacing of Na+-ions to K+- and NH4+-ions is the evidence of a more strong binding of these ions to external mouth of the modified channel compared to the binding of Na+-ions.


The effect of critical operational parameters on the circulation-enhanced electrokinetics.
Electrokinetics (EK) is a technique for soil remediation. However, the acid produced due to the water electrolysis at the anode will cause soil acidification, which may destroy the soil constituents, and reduce contaminant removal efficiency. The formation of a base front produced at the cathode will result in the precipitation of metal hydroxides and a concomitant clogging of pore space. In this study, a circulation-enhanced EK (CEEK) system is designed to neutralize the pH of the working solution and soils for avoiding the above problems. Experiments are conducted by controlling different voltage gradients, electrode materials, and electrode emplacement, respectively. According to the experimental results, the CEEK system could effectively stabilize the current and the pH of processing solution at a neutral range. The strength of voltage gradient is proportional to the current magnitude of the CEEK system. The graphite electrode for CEEK is the better choice than the metal electrodes because graphite electrodes can achieve the lower electricity consumption. The electrode installed in the reservoir without attachment on soils can decrease the pH deviation of the soil matrix.


How We Know What Not To Think.
Humans often represent and reason about unrealized possible actions - the vast infinity of things that were not (or have not yet been) chosen. This capacity is central to the most impressive of human abilities: causal reasoning, planning, linguistic communication, moral judgment, etc. Nevertheless, how do we select possible actions that are worth considering from the infinity of unrealized actions that are better left ignored? We review research across the cognitive sciences, and find that the possible actions considered by default are those that are both likely to occur and generally valuable. We then offer a unified theory of why. We propose that (i) across diverse cognitive tasks, the possible actions we consider are biased towards those of general practical utility, and (ii) a plausible primary function for this mechanism resides in decision making.


Subacute silica nanoparticle exposure induced oxidative stress and inflammation in rat hippocampus combined with disruption of cholinergic system and behavioral functions.
Increasing environmental exposure to silica nanoparticles (SiNPs) and limited neurotoxicity studies pose a challenge for safety evaluation and management of these materials. This study aimed to explore the adverse effects and underlying mechanisms of subacute exposure to SiNPs by the intraperitoneal route on hippocampus function in rats. Data showed that SiNPs induced a significant increase in oxidative/nitrosative stress markers including reactive oxygen species (ROS), malondialdehyde (MDA), protein oxidation (PCO) and nitrite (NO) production accompanied by reduced antioxidant enzyme activity (catalase, superoxide dismutase, and glutathione peroxidase) and decreased glutathione (GSH). Phenotypically, SiNPs exhibited spatial learning and memory impairment in the Morris water maze (MWM) test, a decrease of the discrimination index in the novel object recognition test (NORT) and higher anxiety-like behavior. SiNPs affected the cholinergic system as reflected by reduced acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activity. In addition, SiNPs significantly increased mRNA expression level of genes related to inflammation (TNF-α, IL-1β, IL-6, and COX-2) and decreased mRNA expression level of genes related to cholinergic system including choline acetyltransferase (ChAT), vesicular acetylcholine transporter (VAChT), AChE, muscarinic acetylcholine receptor M1 (m1AChR) and nicotinic acetylcholine receptors (nAChR). Histopathological results further showed an alteration in the hippocampus of treated animals associated with marked vacuolation in different hippocampus areas. These findings provide new insights into the molecular mechanism of SiNPs-induced hippocampal alterations leading to impairment of cognitive and behavioral functions, and implicating oxidative stress and inflammation in the hippocampus, as well as disruption of cholinergic system.


Escherichia coli RecBC helicase has two translocase activities controlled by a single ATPase motor.
E. coli RecBCD is a DNA helicase with two ATPase motors (RecB, a 3'→5' translocase, and RecD, a 5'→3' translocase) that function in repair of double-stranded DNA breaks. The RecBC heterodimer, with only the RecB motor, remains a processive helicase. Here we examined RecBC translocation along single-stranded DNA (ssDNA). Notably, we found RecBC to have two translocase activities: the primary translocase moves 3'→5', whereas the secondary translocase moves RecBC along the opposite strand of a forked DNA at a similar rate. The secondary translocase is insensitive to the ssDNA backbone polarity, and we propose that it may fuel RecBCD translocation along double-stranded DNA ahead of the unwinding fork and ensure that the unwound single strands move through RecBCD at the same rate after interaction with a crossover hot-spot indicator (Chi) sequence.


Carbon dot-based biosensors for the detection of communicable and non -communicable diseases.
Due to their fascinating chemical, optical, electrical, and biological properties carbon dots (CDs or CDots), carbon quantum dots (CQDs), and graphene quantum dots (GQDs) have attracted attention in biosensing as they can greatly improve the detection limit, sensitivity, and selectivity of biosensors. In general, CDs, CQDs, and GQDs are a class of carbon-based nanomaterials that are characterized by extraordinary fluorescence, a size less than 10 nm, high stability, low toxicity, and being easy to synthesize and presenting functional groups in their surface area that vary according to their synthesis source. In this review, a general description of the main methods and precursors reported in the scientific literature for the synthesis of CDs, CQDs, and GQDs are presented, as well as the chemical, optical, electrical, and biological properties that stand out the most from them; moreover, the main objective of this review is to summarize the application of these carbonaceous nanomaterials in biosensors for the detection of communicable and non-communicable diseases. The article summarizes the applications of CDs, CQDs, and GQDs according to the group of diseases they detected using the international classification of diseases in its 10th edition (ICD-10). To facilitate the reader's access to significant information from these biosensors, several tables summarize the information associated with the type of biomarker, the working ranges, and the biosensor assembly.


Experimental and Numerical Investigation of the Mechanical Properties of a Fiber-Reinforced Geopolymer Mortar Blast Resistant Panel.
Geopolymer materials have excellent properties such as high strength, low thermal conductivity, fire resistance, acid and alkali resistance, and low carbon emissions. They can be used as protective engineering materials in places with explosion risks. At present, the common composite blast resistant panel is in the form of a sandwich: the outer layer isgalvanized steel plate, and fiber cement board or calcium carbonate board is used as the inner layer material, as these boards have the advantages of easy installation, good fire resistance, and explosion resistance. This study investigates the effect of adding different types of fibers to geopolymer mortar on the mortar's basic mechanical properties, such as compression strength, bending strength, and impact resistance. The explosive resistance of the fiber-reinforced geopolymer mortar blast resistant panels was evaluated through free-air explosion. In this paper, experimental procedures and numerical simulation have been performed to study the failure modes, maximum deflection, and dynamic response of the fiber-reinforced geopolymer mortar blast resistant panel under free-air explosion. The research results can provide a reference for the design and production of blast resistant panels.


"When patients and families feel abandoned".
Patients with serious illness derive a sense of security by forming strong, healing relationships with their providers. These bonds are particularly strong in life-threatening illnesses, such as cancer, which carry the stigma of death and suffering. These strong relationships create expectations in patients that are not necessarily shared by their clinicians. Providers often focus on treating disease and emphasize technically excellent, "evidence-based" practice while failing to fully appreciate the power of the patient-provider relationship. In contrast, vulnerable patients expect much more than technical competence, including open and clear communication, security, continuity, and access. Patients are often left feeling abandoned when their providers do not meet their expectations, even when their care is technically sound. In this paper, we describe scenarios that can lead to feelings of abandonment and discuss strategies to avoid and respond to them. These strategies can help us maintain healing relationships with our patients by maintaining their trust, confidence, and satisfaction. Cultivating relational aspects of medical practice requires an interchange and takes time. Experienced doctors know this and continue to do so because being present and staying with the patient during difficult times is a pillar of moral and ethical training and a fundamental attribute of a good physician.


Association of dwarfism and floral induction with a grape 'green revolution' mutation.
The transition from vegetative to reproductive growth is an essential process in the life cycle of plants. Plant floral induction pathways respond to both environmental and endogenous cues and much has been learnt about these genetic pathways by studying mutants of Arabidopsis. Gibberellins (GAs) are plant growth regulators important in many aspects of plant growth and in Arabidopsis they promote flowering. Here we provide genetic evidence that GAs inhibit flowering in grapevine. A grapevine dwarf mutant derived from the L1 cell layer of the champagne cultivar Pinot Meunier produces inflorescences along the length of the shoot where tendrils are normally formed. The mutated gene associated with the phenotype is a homologue of the wheat 'green revolution' gene Reduced height-1 (ref. 6) and the Arabidopsis gene GA insensitive (GAI). The conversion of tendrils to inflorescences in the mutant demonstrates that the grapevine tendril is a modified inflorescence inhibited from completing floral development by GAs.


Unexpected findings in trauma patients dying in the intensive care unit: results of 153 consecutive autopsies.
The true incidence of missed injuries in trauma-related deaths is unknown, because in only about 60% of injury-related deaths nationwide is an autopsy performed. Few studies have documented the frequency of missed diagnoses leading to deaths specifically in the trauma ICU population. We attempted to evaluate the incidence and nature of missed injuries and complications in trauma- and burn-related deaths in our ICU given an autopsy rate of close to 100%. The medical records of all trauma- and burn-related deaths in the ICU over a 2-year period were reviewed retrospectively. Missed diagnoses were classified as class 1: major diagnosis that if recognized and treated appropriately might have changed outcomes; class II: major diagnosis that if recognized and treated appropriately would not have changed outcomes; and class III: minor diagnosis. Complete antemortem records were available for 158 patients, of which 153 (97%) underwent autopsy. Mean age was 50 years, and 72% were males. Mean ICU stay was 10 15 days. Four (3%) patients had class I missed diagnoses: bowel infarction, meningitis, retroperitoneal abscess, and bleeding gastric ulcer. Twenty-five (16%) patients had class II diagnoses, and 12 (8%) patients had class III diagnoses. Overall, 81% of 153 patients had either class III diagnoses or no missed injuries or complications. Pneumonia was the most common missed diagnosis. With an autopsy rate of 97%, 3% of deaths bad missed major diagnoses that might have affected outcomes if recognized antemortem. Autopsy findings can still provide valuable feedback in Improving the quality of care of critically ill trauma patients.


Blockade of autophagy reduces pancreatic cancer stem cell activity and potentiates the tumoricidal effect of gemcitabine.
Cancer stem cells (CSCs) are considered responsible for the recurrence and chemoresistance of cancer. Dysregulated autophagy is highly prevalent in many types of cancer including pancreatic cancer and has been implicated in cytoprotection and tumor promotion. This study aimed to investigate the role of autophagy in regulating cancer stemness and chemoresistance of pancreatic cancer. The correlation between autophagy and CSCs and its clinical significance were analyzed using pancreatic cancer tissue microarrays. Genetic and pharmacological approaches were applied to explore the function of autophagy on CSC activity and gemcitabine resistance of pancreatic cancer cells in vitro and in vivo. LC3 expression positively correlated with the expression of CSC markers aldehyde dehydrogenase 1 (ALDH1), CD44, and CD133 in pancreatic cancer tissues. High coexpression of LC3/ALDH1 was associated with both poor overall survival and progression-free survival. In pancreatic cancer cell lines, higher LC3-II expression was observed in the sphere-forming cells than in the bulk cells. Blockade of autophagy by silencing ATG5, ATG7, and BECN1 or the administration of autophagy inhibitor chloroquine markedly reduced the CSC populations, ALDH1 activity, sphere formation, and resistance to gemcitabine in vitro and in vivo. Furthermore, osteopontin (OPN) was found to stimulate LC3-II, ALDH1, CD44, and CD133 expression in PANC-1 cells, whereas this effect could be prevented by OPN knockdown and autophagy blockade. After treatment with various inhibitors against the major signaling pathways downstream of OPN, only the inhibitor of NF-κB activation, BAY 1170-82, could effectively counteract OPN-induced autophagy and CSC activity. According to the histochemical results, pancreatic cancer patients manifesting high levels of OPN/LC3/ALDH1 and OPN/CD44/CD133 had poor survival. Induction of autophagy mediated by OPN/NF-κB signaling is required for maintenance of pancreatic CSC activity. Combination of gemcitabine with pharmacological autophagy inhibitors is a promising therapeutic strategy for pancreatic cancer.


Serotonin decreases HIV-1 replication in primary cultures of human macrophages through 5-HT(1A) receptors.
5-HT (serotonin) is known to be involved in neuroinflammation and immunoregulation. The human immunodeficiency virus (HIV) targets cells such as monocytes/macrophages, which colocalize with 5-HT-releasing cell types, mostly platelets. In this study, we investigated the effects of 5-HT on HIV-1-infected macrophages in vitro. Human macrophages cultured in serum-free medium were treated over 7 days with 5-HT at three concentrations (0.01, 1 and 100 microM) with or without agonists and antagonists of 5-HT(1A) and 5-HT(2) receptors. After 7 days of treatment, macrophages were infected with HIV-1/Ba-L and virus replication was monitored over 16 days and expression of proviral HIV DNA was investigated by PCR after 24 h of infection. Cell surface expression of HIV-1/Ba-L receptor (CD4) and coreceptor (CCR5) was investigated by flow cytometry. The CCR5 ligand, macrophage inflammatory protein-1alpha (MIP-1alpha), was quantified by ELISA in cell culture supernatants and MIP-1alpha mRNA expression was assessed by reverse transcriptase-PCR. In vitro, 5-HT downregulated the membranous expression of CCR5 and led to a decrease of HIV-1 infection, probably through its action on 5-HT(1A) receptors. 5-HT (100 microM) was also able to induce overexpression of MIP-1alpha mRNA leading to an increase of MIP-1alpha secretion by human macrophages. The effects of 5-HT on HIV infection could be a consequence of the increase in MIP-1alpha concentrations and/or CCR5 receptor downregulation. These results suggest that 5-HT can inhibit the replication of HIV-1 in primary culture of human macrophages through its action on 5-HT(1A) receptors.


Simple topological task-based functional connectivity features predict longitudinal behavioral change of fluid reasoning in the RANN cohort.
Recent attention has been given to topological data analysis (TDA), and more specifically persistent homology (PH), to identify the underlying shape of brain network connectivity beyond simple edge pairings by computing connective components across different connectivity thresholds (see Sizemore et al., 2019). In the present study, we applied PH to task-based functional connectivity, computing 0-dimension Betti (B0) curves and calculating the area under these curves (AUC); AUC indicates how quickly a single connected component is formed across correlation filtration thresholds, with lower values interpreted as potentially analogous to lower whole-brain system segregation (e.g., Gracia-Tabuenca et al., 2020). One hundred sixty-three participants from the Reference Ability Neural Network (RANN) longitudinal lifespan cohort (age 20-80 years) were tested in-scanner at baseline and five-year follow-up on a battery of tests comprising four domains of cognition (i.e., Stern et al., 2014). We tested for 1.) age-related change in the AUC of the B0 curve over time, 2.) the predictive utility of AUC in accounting for longitudinal change in behavioral performance and 3.) compared system segregation to the PH approach. Results demonstrated longitudinal age-related decreases in AUC for Fluid Reasoning, with these decreases predicting longitudinal declines in cognition, even after controlling for demographic and brain integrity factors; moreover, change in AUC partially mediated the effect of age on change in cognitive performance. System segregation also significantly decreased with age in three of the four cognitive domains but did not predict change in cognition. These results argue for greater application of TDA to the study of aging.


Transgenic sheep designed for transplantation studies.
Lentiviral vectors have recently emerged as an efficient method of transgene delivery to the germline of animals. We now demonstrate that combining this efficiency with embryo splitting procedures enables the production of monozygotic twins, one of which is transgenic. We propose that this approach can be used to generate animals in which cell or tissue transplantation can be achieved without the use of immunosuppressive regimes.


Cognitive aspects of freezing of gait in Parkinson's disease: a challenge for rehabilitation.
Freezing of gait (FOG) is a very disabling symptom affecting up to half of the patients with Parkinson's disease (PD). Evidence is accumulating that FOG is caused by a complex interplay between motor, cognitive and affective factors, rather than being a pure motor phenomenon. In the current paper, we review the evidence on the specific role of cognitive factors in FOG. Results from behavioral studies show that patients with FOG experience impairments in executive functioning and response selection which predict that motor learning may be compromised. Brain imaging studies strengthen the neural basis of a potential association between FOG and cognitive impairment, but do not clarify whether it is a primary or secondary determinant of FOG. A FOG-related reduction of cognitive resources implies that adaptation of rehabilitation interventions is indicated for patients with FOG to promote the consolidation of learning.


Expression, purification and preliminary characterization of glucagon receptor extracellular domain.
Glucagon is a pancreatic hormone that plays pivotal roles in regulating glucose homeostasis and metabolism. Glucagon exerts its action by binding to its receptor, glucagon receptor (GCGR), one of class B G-protein coupled receptors (GPCRs). Diabetes is a bihormonal disease in which excessive glucagon secretion is a major contributor in the pathogenesis of this disease; elucidation of how glucagon binds to GCGR will facilitate the rational design of the GCGR antagonist for treating diabetic hyperglycemia. Here we report the successful expression and purification of the GCGR extracellular domain (GCGR-ECD) and its fusion protein with the glucagon peptide at its C-terminus (GCGR-ECD-Gc). We utilized the maltose binding protein (MBP) fusion method and disulfide bond isomerase DsbC co-expression approach for the success of the soluble expression of both GCGR-ECD and GCGR-ECD-Gc in Escherichia coli. We also obtained a high yield production of secreted GCGR-ECD with the baculovirus expression system by optimizing its N-terminal secreting signal. We first utilized isothermal titration calorimetry approach to determine the in vitro binding affinities of glucagon to the GCGR-ECD. No significant differences were found between the prokaryotic expressed GCGR-ECD (7.6μM) and the eukaryotic glycosylated one (6.6μM). The observation of the intra ligand-receptor binding within the fusion protein GCGR-ECD-Gc suggests it as a good candidate for further structural study.


Mammalian face as an evolutionary novelty.
The anterior end of the mammalian face is characteristically composed of a semimotile nose, not the upper jaw as in other tetrapods. Thus, the therian nose is covered ventrolaterally by the "premaxilla," and the osteocranium possesses only a single nasal aperture because of the absence of medial bony elements. This stands in contrast to those in other tetrapods in whom the premaxilla covers the rostral terminus of the snout, providing a key to understanding the evolution of the mammalian face. Here, we show that the premaxilla in therian mammals (placentals and marsupials) is not entirely homologous to those in other amniotes; the therian premaxilla is a composite of the septomaxilla and the palatine remnant of the premaxilla of nontherian amniotes (including monotremes). By comparing topographical relationships of craniofacial primordia and nerve supplies in various tetrapod embryos, we found that the therian premaxilla is predominantly of the maxillary prominence origin and associated with mandibular arch. The rostral-most part of the upper jaw in nonmammalian tetrapods corresponds to the motile nose in therian mammals. During development, experimental inhibition of primordial growth demonstrated that the entire mammalian upper jaw mostly originates from the maxillary prominence, unlike other amniotes. Consistently, cell lineage tracing in transgenic mice revealed a mammalian-specific rostral growth of the maxillary prominence. We conclude that the mammalian-specific face, the muzzle, is an evolutionary novelty obtained by overriding ancestral developmental constraints to establish a novel topographical framework in craniofacial mesenchyme.


Clinical, cytogenetic and molecular analyses of a rare case with ring chromosome 15 and review of the literature.
Ring chromosome 15 [r (15)], accompanied by a series of clinical symptoms, is a rare genetic disease. The genotype and phenotypic diversity of patients with r (15) still needed further enrichment. In this study we present a rare case of mosaic ring chromosome 15 with facial anomalies and extremities slenderness. This case involves a 30-year-old woman, unpregnancy within 6 years. Clinical examination of the patient only revealed facial anomalies and extremities slenderness. The result of routine G-band karyotyping was 46,XX,r(15)(p12q26.3)[53]/46,XX,r(15;15)(p11.2q26.3;p11.2q11.2)[28]/45,XX, -15[10]/46,XX,r(15;15)(p11q26.3;p11q26.3)[4]. SNP array was employed to investigate the genome copy number variations (CNVs). The result revealed that there was a micro-duplication of 2.0 Mb at 15q26.3(arr[ph19]15q26.3 (100,400,214- 102,429,112)×3). The duplicated chromosomal section encompassed genes including CHSY1, ALDHIA3, LRRK1, and INS1. We further compared to the cytogenetic characteristics and clinical symptoms of the patient with those already reported by reviewing the literature. This report is especially helpful to supplement the phenotypic diversity of patients with r (15).


Proinsulin-to-C-peptide ratio versus proinsulin-to-insulin ratio in the prediction of incident diabetes: the Insulin Resistance Atherosclerosis Study (IRAS).
Associations of proinsulin-to-insulin ratios with incident type 2 diabetes have been inconsistent. The use of C-peptide as the denominator in the ratio may allow for better prediction because C-peptide concentration is not affected by hepatic insulin clearance. The objective of this paper was to compare fasting intact and split proinsulin-to-insulin ratios (PI/I, SPI/I) with intact and split proinsulin-to-C-peptide ratios (PI/C-pep, SPI/C-pep) in the prediction of type 2 diabetes. Prospective data on 818 multi-ethnic adults without diabetes at baseline from the Insulin Resistance Atherosclerosis Study (IRAS) were used. Insulin sensitivity (S(I)) and acute insulin response (AIR) were determined from frequently sampled intravenous glucose tolerance tests, and fasting intact and split proinsulin were measured using specific two-site monoclonal antibody-based immunoradiometric assays. Associations of proinsulin ratios with type 2 diabetes were determined using logistic regression and differences in prediction were assessed by comparing areas under the receiver operating characteristic curve (AROCs). In logistic regression analyses, PI/C-pep and SPI/C-pep were more strongly associated with incident type 2 diabetes (n = 128) than PI/I and SPI/I, and were significantly better predictors of diabetes in AROC analyses (PI/C-pep = 0.662 vs PI/I = 0.603, p = 0.02; SPI/C-pep = 0.690 vs SPI/I = 0.631, p = 0.01). Both PI/C-pep and SPI/C-pep were associated with type 2 diabetes after adjustment for age, sex, ethnicity, waist circumference, impaired glucose tolerance, lipids and S(I). Both PI/C-pep and SPI/C-pep were significantly associated with incident type 2 diabetes in models that included AIR. Proinsulin-to-C-peptide ratios were stronger predictors of diabetes in comparison with proinsulin-to-insulin ratios. These findings support the use of C-peptide as the denominator for proinsulin ratios, to more accurately reflect the degree of disproportional hyperproinsulinaemia.


Proton potential-dependent polyamine transport by vacuolar membrane vesicles of Saccharomyces cerevisiae.
Vacuolar membrane vesicles of Saccharomyces cerevisiae accumulated spermine and spermidine in the presence of ATP, not in the presence of ADP. Spermine and spermidine transport at pH 7.4 showed saturation kinetics with Km values of 0.2 mM and 0.7 mM, respectively. Spermine uptake was competitively inhibited by spermidine and putrescine, but was not affected by seven amino acids, substrates of active transport systems of vacuolar membrane. Spermine transport was inhibited by the H(+)-ATPase-specific inhibitors bafilomycin A1 and N,N'-dicyclohexylcarbodiimide, but not by vanadate. It was also sensitive to Cu2+ or Zn2+ ions, inhibitors of vacuolar H(+)-ATPase. Both 3,5-di-tert-butyl-4-hydroxybenzilidenemalononitrile (SF6847) and nigericin blocked completely the spermine uptake, but valinomycin did not. [14C]Spermine accumulated in the vesicles was exchangeable with unlabeled spermine and spermidine. However, it was released by a protonophore only in the presence of a counterion such as Ca2+. These results indicate that a polyamine-specific transport system depending on a proton potential functions in the vacuolar membrane of this organism.


Cloning and expression of odorant receptors.
Myriads of odorous molecules that vary widely in structure are nevertheless readily detected and discriminated by the sense of smell, but how this is achieved by the olfactory system has been a long-standing puzzle. Several different models have been proposed, and previous observations indicate that the recognition sites for odorous molecules could be G-protein-coupled receptor proteins, an idea supported by the discovery of a new gene family that probably encodes a diversity of odorant receptors. Here we report the identification of new members of the gene family encoding putative odorant receptors and demonstrate that they are indeed transcribed in olfactory receptor neurons. Furthermore, the receptor-encoding complementary DNA is expressed in non-neuronal surrogate cells, which generate second messenger responses upon stimulation with appropriate odorants, indicating that the receptors recognize odorants and couple to G proteins of the host cells.


Aquaporin 1 promotes sensitivity of anthracycline chemotherapy in breast cancer by inhibiting β-catenin degradation to enhance TopoIIα activity.
Anthracyclines are a class of conventional and commonly used frontline chemotherapy drugs to treat breast cancer. However, the anthracycline-based regimens can only reduce breast cancer mortality by 20-30%. Furthermore, there is no appropriate biomarker for predicting responses to this kind of chemotherapy currently. Here we report our findings that may fill this gap by showing the AQP1 (Aquaporin1) protein as a potential response predictor in the anthracycline chemotherapy. We showed that breast cancer patients with a high level of AQP1 expression who underwent the anthracycline treatment had a better clinical outcome relative to those with a low level of AQP1 expression. In the exploration of the underlying mechanisms, we found that the AQP1 and glycogen synthase kinase-3β (GSK3β) competitively interacted with the 12 armadillo repeats of β-catenin, followed by the inhibition of the β-catenin degradation that led to β-catenin's accumulation in the cytoplasm and nuclear translocation. The nuclear β-catenin interacted with TopoIIα and enhanced TopoIIα's activity, which resulted in a high sensitivity of breast cancer cells to anthracyclines. We also found, the miR-320a-3p can attenuate the anthracycline's chemosensitivity by inhibiting the AQP1 expression. Taken together, our findings suggest the efficacy of AQP1 as a response predictor in the anthracycline chemotherapy. The application of our study includes, but is not limited to, facilitating screening of the most appropriate breast cancer patients (who have a high AQP1 expression) for better anthracycline chemotherapy and improved prognosis purposes.


Challenges in the Measurement and Interpretation of Serum Titanium Concentrations.
The measurement of circulating metal ion levels in total hip arthroplasty patients continues to be an area of clinical interest. National regulatory agencies have recommended measurement of circulating cobalt and chromium concentrations in metal-on-metal bearing symptomatic total hip arthroplasty patients. However, the clinical utility of serum titanium (Ti) measurements is less understood due to wide variations in reported values and methodology. Fine-scale instrumentation for detecting in situ Ti levels continues to improve and has transitioned from graphite furnace atomic absorption spectroscopy to inductively coupled plasma optical emission spectrometry or inductively coupled plasma mass spectrometry. Additionally, analytical interferences, variable sample types, and non-standardized sample collection methods complicate Ti measurement and underlie the wide variation in reported levels. Normal reference ranges and pathologic ranges for Ti levels remain to be established quantitatively. However, before these ranges can be recognized and implemented, methodological standardization is necessary. This paper aims to provide background and recommendations regarding the complexities of measurement and interpretation of circulating Ti levels in total hip arthroplasty patients.


Evaluation of regional cerebral blood flow with 99mTc-d, 1 HM-PAO and SPECT.
The imaging of regional cerebral blood flow (rCBF) by 99mTc-d, 1 HM-PAO and SPECT is described. Its relevance to clinical syndromes, such as stroke, transient ischemic attacks, various forms of dementia, epilepsy, and subarachnoid hemorrhage, is reported.


Cooperative coupling and role of heme a in the proton pump of heme-copper oxidases.
In the last few years, evidence has accumulated supporting the applicability of the cooperative model of proton pumps in cytochrome systems, vectorial Bohr mechanisms, to heme-copper oxidases. The vectorial Bohr mechanism is based on short- and long-range protonmotive cooperative effects linked to redox transitions of the metal centers. The crystal structure of oxidized and reduced bovine-heart cytochrome c oxidase reveals, upon reduction, the occurrence of long-range conformational changes in subunit I of the oxidase. Analysis of the crystal structure of cytochrome c oxidase shows the existence of hydrogen-bonded networks of amino acid residues which could undergo redox-linked pK shifts resulting in transmembrane proton translocation. Our group has identified four proteolytic groups undergoing reversible redox-linked pK shifts. Two groups result in being linked to redox transitions of heme a3. One group is apparently linked to CuB. The fourth group is linked to oxido-reduction of heme a. We have shown that the proton transfer resulting from the redox Bohr effects linked to heme a and CuB in the bovine oxidase displays membrane vectorial asymmetry, i.e., protons are taken up from the inner aqueous space (N), upon reduction, and released in the external space (P), upon oxidation of the metals. This direction of proton uptake and release is just what is expected from the vectorial Bohr mechanism. The group linked to heme a, which can transfer up to 0.9 H+/e- at pHs around neutrality, can provide the major contribution to the proton pump. It is proposed that translocation of pumped protons, linked to electron flow through heme a, utilizes a channel (channel D) which extends from a conserved aspartate at the N entrance to a conserved glutamate located between heme a and the binuclear center. The carboxylic group of this glutamic acid, after having delivered, upon electron flow through heme a, pumped protons towards the P phase, once reprotonated from the N phase, moves to deliver, subsequently, to the binuclear center chemical protons consumed in the conversion of the peroxy to ferryl and of the latter to the oxy intermediate in the redox cycle. Site-directed mutagenesis of protolytic residues in subunit I of the aa3-600 quinol oxidase of Bacillus subtilis to non-polar residues revealed that the conserved Lys 304 is critical for the proton pumping activity of the oxidase. Crystal structures of cytochrome c oxidase show that this lysine is at the N entrance of a channel which translocates the protons consumed for the production of the peroxy intermediate. Inhibition of this pathway, by replacement of the lysine, short-circuits protons from channel D to the binuclear center, where they are utilized in the chemistry of oxygen reduction.


Evolutionary convergence or homology? Comparative cytogenomics of Caesalpinia group species (Leguminosae) reveals diversification in the pericentromeric heterochromatic composition.
We demonstrated by cytogenomic analysis that the proximal heterochromatin of the Northeast Brazilian species of Caesalpinia group is enriched with phylogenetically conserved Ty3/Gypsy-Tekay RT, but diverge in the presence of Ty3/Gypsy-Athila RT and satDNA. The Caesalpinia Group includes 225 species and 27 monophyletic genera of which four occur in Northeastern Brazil: Erythrostemon (1 sp.), Cenostigma (7 spp.), Libidibia (1 sp.), and Paubrasilia (1 sp.). The last three genera are placed in different clades in the Caesalpinia Group phylogeny, and yet they are characterized by having a numerically stable karyotype 2n = 24 (16 M+8A) and GC-rich heterochromatic bands (chromomycin A3 positive/CMA+ bands) in the proximal chromosome regions. To characterize the composition of their heterochromatin and test for the homology of these chromosomal regions, genomic DNA was extracted from Cenostigma microphyllum, Libidibia ferrea, and Paubrasilia echinata, and sequenced at low coverage using the Illumina platform. The genomic repetitive fractions were characterized using a Galaxy/RepeatExplorer-Elixir platform. The most abundant elements of each genome were chromosomally located by fluorescent in situ hybridization (FISH) and compared to the CMA+ heterochromatin distribution. The repetitive fraction of the genomes of C. microphyllum, L. ferrea, and P. echinata were estimated to be 41.70%, 38.44%, and 72.51%, respectively. Ty3/Gypsy retrotransposons (RT), specifically the Tekay lineage, were the most abundant repeats in each of the three genomes. FISH mapping revealed species-specific patterns for the Tekay elements in the proximal regions of the chromosomes, co-localized with CMA+ bands. Other species-specific patterns were observed, e.g., for the Ty3/Gypsy RT Athila elements which were found in all the proximal heterochromatin of L. ferrea or restricted to the acrocentric chromosomes of C. microphyllum. This Athila labeling co-localized with satellite DNAs (satDNAs). Although the Caesalpinia Group diverged around 55 Mya, our results suggest an ancestral colonization of Tekay RT in the proximal heterochromatin. Thus, the present-day composition of the pericentromeric heterochromatin in these Northeast Brazilian species is a combination of the maintenance of an ancestral Tekay distribution with a species-specific accumulation of other repeats.


Use of contraceptives prior to and after conception and exposure to other fetal hazards.
In a large prospective study, set up to determine whether reproductive outcomes are affected by prior contraceptive use, 34,344 women were recruited at their first antenatal visit and followed until pregnancy termination. This paper summarizes the study methods and describes the demographic differences between subjects who used various contraceptives shortly before conception, and those who experienced contraceptive failures. Thirty percent of the women had used oral contraceptives (OCs) during the 5 months prior to conception and 2.4% had continued using them after their LMP. IUDs had been used by 5.8% of women before conception and 1.1% after. If these rates are also true for the whole population of the U.S., they indicate that approximately 70,000 babies are born each year following pill-failures and 30,000 following failures of IUD's. Exposure to other fetal hazards was common, with 48% of pregnant women drinking alcohol, 28% smoking, and 15% receiving diagnostic X-rays. OC use was more common among smokers and drinkers, and OC failures were significantly associated with the use of anti-epileptics, aspirin, and with exposure to radiation.


Significance of Aspergillus fumigatus isolation from respiratory specimens of nongranulocytopenic patients.
The aim of this study was to determine the significance of isolation of Aspergillus fumigatus from cultures of respiratory specimens in nongranulocytopenic patients. The medical records of patients with respiratory specimens positive for Aspergillus fumigatus who were admitted to an adult pneumology ward were reviewed during a 2-year period. A total of 80 respiratory specimens from 76 patients yielded Aspergillus fumigatus. Forty-eight patients were colonized with Aspergillus fumigatus, whereas the 28 (37%) remaining patients had pulmonary aspergillosis, manifest as aspergilloma ( n=19 patients), chronic necrotizing pulmonary aspergillosis ( n=7 patients), and bronchial aspergillosis ( n=2 patients). The presence of typical hyphae in direct examination of bronchoscopic specimens was more likely to be found in infected than in colonized patients ( P=0.04). No immunological test was positive in colonized patients, whereas anti- Aspergillus antibodies were detected in 55% of infected patients ( P<0.001). Pulmonary tuberculosis was the most common underlying lung disease in patients with aspergilloma, but it was not found in any patient with chronic necrotizing pulmonary aspergillosis ( P=0.006). Anti- Aspergillus antibodies were more likely to be detected in patients with aspergilloma (78%) than in patients with chronic necrotizing pulmonary aspergillosis (14%) ( P=0.007). The analysis of predisposing factors, in conjunction with immunological tests and examination of bronchoscopic specimens, is helpful in distinguishing between colonization and infection with Aspergillus fumigatus, as well as for differentiating between aspergilloma and chronic necrotizing pulmonary aspergillosis.


Pseudo-Ligandless Click Chemistry for Oligonucleotide Conjugation.
Particularly for its use in bioconjugations, the copper-catalyzed (or copper-promoted) azide-alkyne cycloaddition (CuAAC) reaction or 'click chemistry', has become an essential component of the modern chemical biologist's toolbox. Click chemistry has been applied to DNA, and more recently, RNA conjugations, and the protocols presented here can be used for either. The reaction can be carried out in aqueous buffer, and uses acetonitrile as a minor co-solvent that serves as a ligand to stabilize the copper. The method also includes details on the analysis of the reaction product. © 2016 by John Wiley & Sons, Inc.


Evaluation of mechanical strength and bone regeneration ability of 3D printed kagome-structure scaffold using rabbit calvarial defect model.
In clinical conditions, the reconstructions performed in the complex and three-dimensional bone defects in the craniomaxillofacial (CMF) area are often limited in facial esthetics and jaw function. Furthermore, to regenerate a bone defect in the CMF area, the used scaffold should have unique features such as different mechanical strength or physical property suitable for complex shape and function of the CMF bones. Therefore, a three-dimensional synthetic scaffold with a patient-customized structure and mechanical properties is more suitable for the regeneration. In this study, the customized kagome-structure scaffold with complex morphology was assessed in vivo. The customized 3D kagome-structure model for the defect region was designed according to data using 3D computed tomography. The kagome-structure scaffold and the conventional grid-structure scaffold (as a control group) were fabricated using a 3D printer with a precision extruding deposition head using poly(ε-caprolactone) (PCL). The two types of 3D printed scaffolds were implanted in the 8-shaped defect model on the rabbit calvarium. To evaluate the osteoconductivity of the implanted scaffolds, new bone formation, hematoxylin and eosin staining, immunohistochemistry, and Masson's trichrome staining were evaluated for 16 weeks after implantation of the scaffolds. To assess the mechanical robustness and stability of the kagome-structure scaffold, numerical analysis considering the 'elastic-perfectly plastic' material properties and deformation under self-contact condition was performed by finite element analysis. As a result, the kagome-structure scaffold fabricated using 3D printing technology showed excellent mechanical robustness and enhanced osteoconductivity than the control group. Therefore, the 3D printed kagome-structure scaffold can be a better option for bone regeneration in complex and large defects than the conventional grid-type 3D printed scaffold.


Cost-effectiveness of edoxaban vs low-molecular-weight heparin and warfarin for cancer-associated thrombosis in Brazil.
Venous thromboembolism (VTE) is the second leading cause of death in cancer patients. In Brazil, even though low-molecular-weight heparin (LMWH) is the gold standard of care for the management of cancer-associated thrombosis (CAT), its cost limits its use and therefore warfarin is commonly prescribed. Direct oral anticoagulants (DOACs), such as edoxaban, have been introduced as an alternative in this setting. The aim of this study was to compare the cost-effectiveness of edoxaban with LMWH (Model 1) and warfarin (Model 2) to support clinicians and hospitals when choosing an anticoagulant to manage CAT. Cost-effectiveness analyses were performed using Markov state-transition models over a timeframe of 5 years, in a hypothetical, 64 years-old patients cancer population with an index VTE event. Transition probabilities, costs, quality-adjusted life years (QALYs) and risk reductions were either derived from the literature, estimated or calculated. A willingness-to-pay limit of 3 Gross Domestic Product (GDP) per head was used. Deterministic and probabilistic sensitivity analyses were performed for robustness. The main outcome of this study was the incremental cost-effectiveness ratio (ICER), expressed as cost per QALY gained. Model 1 base case analysis demonstrated dominance of edoxaban compared to LMWH, with an ICER of $5204.46, representing cost saved per QALY lost. In Model 2, edoxaban was associated with a $736.90 cost increase vs. warfarin, with an ICER of $2541.03. Sensitivity analyses confirmed base-case results. Edoxaban represents a cost-saving alternative to LMWH for the management of CAT and is cost-effective vs. warfarin.


Labor and delivery outcomes among young adolescents.
We sought to determine whether young adolescents aged 11-14 years and teens aged 15-17 and 18-19 years have an increased risk of cesarean or operative delivery, as well as maternal or neonatal delivery-related morbidity, compared to young adults aged 20-24 years. We conducted a retrospective population-based cohort study using Washington State birth certificate data linked to hospital records from 1987 through 2009 for 26,091 nulliparas with singleton gestations between 24-43 weeks. We compared young adolescents aged 11-14 years, young teens aged 15-17 years, and older teens aged 18-19 years to young adults aged 20-24 years. The primary outcome was method of delivery. Secondary outcomes included postpartum hemorrhage, shoulder dystocia, third- and fourth-degree perineal lacerations, chorioamnionitis, prolonged maternal length of stay, gestational age at delivery, birthweight, respiratory distress syndrome, neonatal length of stay, and death. We used multivariate regression to assess associations between age and delivery outcomes. Young adolescents aged 11-14 years had a lower risk of cesarean (risk ratio [RR], 0.73; 95% confidence interval [CI], 0.65-0.83) and operative vaginal (RR, 0.87; 95% CI, 0.78-0.97) delivery compared to young adults aged 20-24 years. Compared to young adults, young adolescents had an increased risk of prolonged length of stay for both vaginal and cesarean delivery (RR, 1.34; 95% CI, 1.20-1.49, and RR, 1.71; 95% CI, 1.38-2.12, respectively), with no significant differences in indication for cesarean delivery or other measures of maternal morbidity. Young adolescents had an increased risk of preterm delivery (RR, 2.11; 95% CI, 1.79-2.48), low and very low birthweight (RR, 2.08; 95% CI, 1.73-2.50, and RR, 3.25; 95% CI, 2.22-4.77, respectively), and infant death (RR, 3.90; 95% CI, 2.36-6.44) compared to young adults. Young adolescents have a decreased risk of cesarean and operative vaginal delivery compared to young adults; however, their neonates face higher risks of preterm delivery, low and very low birthweight, and death. This information can be used to inform clinical care for this population.


Role of Instruction Adherence During Highly Structured Robotic Arm Training on Motor Outcomes for Individuals After Chronic Stroke.
The aim of this study was to examine the effects of instruction adherence on upper limb motor outcomes after highly structured intervention. A secondary data analysis was completed using mixed linear modeling design. Thirty chronic stroke survivors with moderate-to-severe arm impairment were assigned explicit movement instructions and completed 12 robotic therapy sessions for 4 wks. Instruction adherence was measured weekly using the Manipulation Check Questionnaire. Motor outcomes were measured at baseline, discharge, and at 4-wk follow-up using the Fugl-Meyer Assessment-Upper Extremity and Wolf Motor Function Test-Functional Ability Scale. There were no effects of adherence-related variables on upper limb motor outcomes (P > 0.2). Participant scores improved for both Fugl-Meyer Assessment-Upper Extremity and Wolf Motor Function Test-Functional Ability Scale from baseline to follow-up (P < 0.001). Participants improved motor function after 12 wks of robotic training; however, instruction adherence did not affect motor improvement. Adherence to explicit information may play a limited role in motor learning for stroke survivors with moderate-to-severe arm impairment during highly structured training protocols.


Presenile primary cognitive decline or Alzheimer's dementia: 7-year clinical and neuropsychological follow-up.
Early diagnosis of presenile Alzheimer's disease (AD), which would serve for prognosis and for guiding choices of treatment, is still an important, difficult task for the clinical neurologist. We studied 24 patients, 12 of whom had minor cognitive impairment or questionable dementia (PICD) and 12 who met NINCDS-ADRDA criteria for presenile AD (PAD). Using clinical, neuropsychological, neurophysiological and neuroradiological methods, we followed the patients up to two disease end-points: death or untestable condition. This paper concentrates on the main clinical and neuropsychological findings relative to these two end-points. All PAD patients evolved into clinically evident Alzheimer-type dementia, became untestable within 60 months and died within 72 months. Only 3 of the PICD patients became demented; 2 of them died during the follow-up and 1 died eight months later. The other 9 PICD patients showed only moderate cognitive decline, compatible with normal aging processes. Neurophysiological and neuroradiological findings might be an important tool for arriving at a correct early diagnosis, when they are assessed with clinical neuropsychological data.


Arthroscopic versus mini-open rotator cuff repair: A meta-analysis.
An all-arthroscopic rotator cuff repair (ASR) may result in less postoperative pain and better functional outcomes than the mini-open (MOR) approach. This meta-analysis provides an updated assessment of the current literature which compares the clinical outcomes of mini-open versus all arthroscopic rotator cuff repair techniques. The main online databases were accessed in October 2021. All the trials directly comparing primary ASR versus MOR for rotator cuff rupture were accessed. Studies concerning revision settings were not eligible, nor where those combining the surgical procedures with other adjuvants. A total of 21 articles were retrieved. Data from 1644 procedures (ASR = 995, MOR = 649) were collected. The mean follow-up was 26.7 (6.0-56.4) months. Comparability was found between ASR and MOR groups at baseline with regards to age (P = 0.3), gender (P = 0.7) and mean duration of the follow-up (P = 0.7). No difference was found between ASR and MOR with regard to surgical duration (P = 0.05), Constant score (P = 0.2), University of California at Los Angeles Shoulder (P = 0.3), American Shoulder and Elbow Surgeons Shoulder (P = 0.5), VAS (P = 0.2), forward flexion (P = 0.3), abduction (P = 0.3), external rotation (P = 0.2), internal rotation (P = 0.7), re-tear (P = 0.9), adhesive capsulitis (P = 0.5). Arthroscopic and mini-open rotator cuff repair result in similar clinical outcomes. Male gender and older age lead to greater rates of rotator cuff re-tears, while longer surgical duration was associated with a greater rate of adhesive capsulitis.


Respiratory arrest outside hospital.
Of 954 attempted resuscitations outside hospital performed by ambulance personnel, 48 patients (5%) had primary respiratory arrest. Comparing this group with those manifesting cardiorespiratory arrest, patients with primary respiratory arrest were significantly more likely to be female (25 of 48 vs 269 of 906-P < 0.005), were more likely to have a non-cardiac cause (67% vs. 22%-P < 0.00001) and more likely to have witnessed arrest. Of all arrests witnessed by ambulance crew, 35% were respiratory arrests. Basic and advanced life-support was delivered sooner. Outcome was significantly better, with 19 patients (40%) being discharged compared to only 49 patients (5.1%) discharged in cases of cardiorespiratory arrest (p < 0.00001). Considering that many respiratory arrests were witnessed by ambulance crew, the type of crew (EMT or paramedic) made no difference to outcome. Our findings suggest that patients manifesting respiratory arrest outside hospital are a heterogeneous group who have a relatively good prognosis regardless of the type of ambulance crew that attends.


The Effect of the Video Assistant Referee System Implementation on Match Physical Demands in the Spanish LaLiga.
The present study aimed to analyze the influence of the Video Assistant Referee (VAR) on match physical demands in the top Spanish professional football league. Match physical demand data from all the matches for two seasons (2017/2018 and 2018/2019) in the First Spanish Division (n = 1454) were recorded using an optical tracking system (ChyronHego®). Total distance, relative total distance covered per minute, distance covered between 14-21 km·h-1, distance covered between 21-24 km·h-1, and distance covered at more than 24 km·h-1 were analyzed; also, the number of sprints between 21-24 km·h-1 and more than 24 km·h-1 were taken into consideration. The times the VAR intervened in matches were also taken into account. Results showed that total distance and relative total distance significantly decreased in seasons with VAR compared to seasons without VAR. Finally, distance covered between 21-24 km·h-1, distance covered at more than 24 km·h-1, and the number of high-intensity efforts between 21-24 km·h-1 and more than 24 km·h-1 increased in seasons with VAR compared to seasons without VAR, but the differences were nonsignificant. Thus, these findings help practitioners to better understand the effects of the VAR system on professional football physical performance and to identify strategies to reproduce competition demands.


Enhancing the antitumor activity of ErbB blockade with histone deacetylase (HDAC) inhibition.
Molecular inhibition of the ErbB signaling pathway represents a promising cancer treatment strategy. Preclinical studies suggest that enhancement of antitumor activity can be achieved by maximizing ErbB signaling inhibition. Using cDNA microarrays, we identified histone deacetylase (HDAC) inhibitors as having strong potential to enhance the effects of anti-ErbB agents. Studies using a 20,000 element (20K) cDNA microarray demonstrate decreased transcript expression of ErbB1 (epidermal growth factor receptor) and ErbB2 in DU145 (prostate) and ErbB2 in SKBr3 (breast) cancer cell lines. Additional changes in the DU145 gene expression profile with potential interaction to ErbB signaling include down-regulation of caveolin-1 and hypoxia inducible factor 1-alpha (HIF1-alpha), and up-regulation of gelsolin, p19(INK4D) and Nur77. Findings were validated using real time RT-PCR and Western blot analysis. Enhanced proliferative inhibition, apoptosis induction and signaling inhibition were demonstrated when combining HDAC inhibition with ErbB blockade. These results suggest that used cooperatively, anti-ErbB agents and HDAC inhibitors may offer a promising strategy of dual targeted therapy. Additionally, microarray data suggest that the beneficial interaction of these agents may not derive solely from modulation of ErbB expression, but may result from effects on other oncogenic processes including angiogenesis, invasion and cell cycle kinetics.


Low conversion rate of ocular to generalized myasthenia gravis in Singapore.
Ocular myasthenia gravis (OMG) is a common condition of the neuromuscular junction that may convert to generalized myasthenia gravis (GMG). Our aim in this study was to determine the conversion rate and predictive factors for generalization in OMG, in an Asian population. The investigation consisted of a retrospective study of OMG patients with a minimum 2 years of follow-up. Among 191 patients with OMG, 155 had the minimum 2-year follow-up. The conversion rate at median follow-up (40.8 months) was 10.6% (95% confidence interval 7.9%-13.3%), and at the 2-year follow-up it was 7.7% (95% confidence interval 5.6%-9.8%). At baseline, the predictive factors for generalization were positive acetylcholine receptor antibodies (hazard ratio 3.71, P = 0.024), positive repetitive nerve stimulation (RNS) studies (hazard ratio 4.42, P = 0.003), and presence of radiologically presumed or pathologically confirmed thymoma (hazard ratio 3.10, P = 0.013). The conversion rate of OMG to GMG in Asian patients is low, as predicted by presence of acetylcholine receptor antibodies, presence of thymoma, and positive RNS studies. Muscle Nerve 57: 756-760, 2018.


Long-term trends and speciation of artificial radionuclides in two submerged macrophytes of the Yenisei River: A comparative study of Potamogeton lucens and Fontinalis antipyretica.
Long-term trends of artificial radionuclides, from 2003 to 2018, in two abundant species of macrophytes, shining pondweed, Potamogeton lucens, and water moss, Fontinalis antipyretica, have been analyzed to estimate the indicative reliability of these two species as biomonitors of radioactive contamination in a river system and to quantify the decrease in the content of artificial radionuclides in the Yenisei River. Time-dependent trends of artificial radionuclides in the biomass of these species were similar, resulting in estimates of effective half-lives for 54Mn, 58Co, 60Co, 65Zn, 137Cs, and 152Eu similar for both species. Concentrations of artificial radionuclides in biomass of shining pondweed and water moss correlated with annual discharges of the radionuclides to the Yenisei at different levels of significance, and the strongest (R2 > 0.7) positive correlation (p < 0.05) was obtained for 60Co, 65Zn, and 152Eu. Concentrations of 60Co, 137Cs, and 152Eu in water moss were 2-7.5 times higher than in shining pondweed, and considerable percentages of those isotopes were recorded in extracellular particulate matter, which was largely represented by epiphytic diatoms. Higher concentrations of artificial radionuclides in the biomass of water moss can be considered as an advantage of water moss as a monitor of radioactive contamination of the Yenisei, while shining pondweed is more useful for estimation of annual deposits of radionuclides in vegetation of the Yenisei and spatial transfer of radionuclides downstream of the discharge site. Despite differences in concentrations of artificial radionuclides, both species can be considered as reliable indicators of radioactive contamination of the river on a long-term scale.


The histone methyltransferase SMYD2 methylates PARP1 and promotes poly(ADP-ribosyl)ation activity in cancer cells.
Poly(ADP-ribose) polymerase-1 (PARP1) catalyzes the poly(ADP-ribosyl)ation of protein acceptors using NAD(+) as the substrate is now considered as an important target for development of anticancer therapy. PARP1 is known to be post-translationally modified in various ways including phosphorylation and ubiquitination, but the physiological role of PARP1 methylation is not well understood. Herein we demonstrated that the histone methyltransferase SMYD2, which plays critical roles in human carcinogenesis, mono-methylated PARP1. We confirmed lysine 528 to be a target of SMYD2-dependent PARP1 methylation by LC-MS/MS and Edman Degradation analyses. Importantly, methylated PARP1 revealed enhanced poly(ADP-ribose) formation after oxidative stress, and positively regulated the poly(ADP-ribosyl)ation activity of PARP1. Hence, our study unveils a novel mechanism of PARP1 in human cancer through its methylation by SMYD2.


A new bulking agent (polyacrylamide hydrogel) for treating stress urinary incontinence in women.
To evaluate the efficacy and safety of a transurethral injection with polyacrylamide hydrogel (PAHG) for treating stress urinary incontinence (SUI) in women. The study comprised 17 women with pure SUI and eight with mixed incontinence (age range 35-84 years). Between November 2001 and November 2003, the women were injected into the urethral wall with a PAHG (Aquamid, Contura International A/S, Denmark), a homogenous, highly biocompatible, atoxic, nonresorbable sterile watery gel consisting of approximately 2.5% cross-linked polyacrylamide and 97.5% nonpyrogenic water. The women were followed prospectively (assessed at 1, 3 and 12 months after treatment) and their clinical outcome documented. Four women did not complete the study, and of the remainder, eight (38%) were subjectively dry and a further nine (43%) improved. Objectively, urine leakage/24 h was decreased by 93% and the number of incontinence episodes by 87%. Quality-of-life measures improved significantly in all domains other than general health perception. There were no significant changes of urodynamic variables. Treatment-related adverse events were recorded in 16 women. Urinary tract infection (10 cases) and urinary retention (five cases) were most common. There were no injection site reactions. PAGH seems to be a promising new bulking agent for the treatment of SUI.


Slow and Sustained Release of Carbonate Ions from Amino Acids for Controlled Hydrothermal Growth of Alkaline-Earth Carbonate Single Crystals.
Alkaline-earth metal carbonate materials have attracted wide interest because of their high value in many applications. Various sources of carbonate ions (CO3 2-), such as CO2 gas, alkaline-metal carbonate salts, and urea, have been reported for the synthesis of metal carbonate crystals, yet a slow and sustained CO3 2- release approach for controlled crystal growth is much desired. In this paper, we demonstrate a new chemical approach toward slow and sustained CO3 2- release for hydrothermal growth of large alkaline-earth metal carbonate single crystals. Such an approach is enabled by the multiple hydrolysis of a small basic amino acid (arginine, Arg). Namely, the amino groups of Arg hydrolyze to form OH- ions, making the solution basic, and the hydrolysis of the guanidyl group of Arg is hydrothermally triggered to produce urea and ammonia, followed by the hydrolysis of urea to produce CO2 and ammonia and then the release of CO3 2- because of the reaction between CO2 and the OH- ions hydrolyzed from ammonia. Such a CO3 2- release behavior enables the slow and controlled growth of various carbonate single crystals over a wide range of pH values. The growth of uniform rhombohedron MgCO3 single crystals with variable morphologies and crystal sizes is studied in detail. The influences of reaction temperature, solution pH, precursor type, and concentration on the morphology and size of the resulting MgCO3 crystals are elucidated. The crystal evolution mechanism is also proposed and discussed with various supportive data.


Time-resolved fluorescence spectroscopy of lumazine protein from Photobacterium phosphoreum using synchrotron radiation.
Time-resolved fluorescence on lumazine protein from Photobacterium phosphoreum was performed with synchrotron radiation as a source of continuously tunable excitation. The experiments yielded structural and dynamic details from which two aspects became apparent. From fluorescence anisotropy decay monitoring of lumazine fluorescence with different excitation wavelengths, the average correlation times were shown to change, which must indicate the presence of anistropic motion of the protein. A similar study with 7-oxolumazine as the fluorescent ligand led to comparable results. The other remarkable observation dealt with the buildup of acceptor fluorescence, also observed with 7-oxolumazine although much less pronounced, which is caused by the finite energy transfer process between the single donor tryptophan and the energy accepting lumazine derivatives. Global analytical approaches in data analysis were used to yield realistic correlation times and reciprocal transfer rate constants. It was found that the tryptophan residue has a large motional freedom as also reported previously for this protein and for the related protein from P. leiognathi (Lee et al. 1985; Kulinski et al. 1987). The average distance between the tryptophan residue and the ligand donor-acceptor couple has been determined to be 2.7 nm for the same donor and two different acceptors.


TDP-43 inhibitory peptide alleviates neurodegeneration and memory loss in an APP transgenic mouse model for Alzheimer's disease.
Alzheimer's disease (AD) is the leading cause of dementia in the elderly, characterized clinically by progressive decline in cognitive function and neuropathologically by the presence of senile plaques and neuronal loss in the brain. While current drugs for AD are always employed as symptomatic therapies with variable benefits, there is no treatment to delay its progression or halt neurodegeneration. TAR DNA-binding protein 43 (TDP-43) proteinopathy has increasingly been implicated as a prominent histopathological feature of AD and related dementias. Our recent studies have implicated mitochondria as critical targets of TDP-43 neurotoxicity. Here, we demonstrate that the suppression of mitochondrial-associated TDP-43 protects against neuronal loss and behavioral deficits in 5XFAD transgenic mice recapitulating AD-related phenotypes. In AD patients and 5XFAD mice, the level of TDP-43 is increased in mitochondria, and TDP-43 highly co-localizes with mitochondria in brain neurons exhibiting TDP-43 proteinopathy. Chronic administration of a TDP-43 mitochondrial localization inhibitory peptide, PM1, significantly alleviates TDP-43 proteinopathy, mitochondrial abnormalities, microgliosis and even neuronal loss without effect on amyloid plaque load in 12-month-old 5XFAD mice well after the onset of symptoms. Additionally, PM1 also improves the cognitive and motor function in 12-month-old 5XFAD mice and completely prevents the onset of mild cognitive impairment in 6-month-old 5XFAD mice. These data indicate that mitochondria-associated TDP-43 is likely involved in AD pathogenesis and that the inhibitor of mitochondria-associated TDP-43 may be a valuable drug to treat underlying AD.


[Comparative study of artificial circulation for the liver after cardiogenic shock: pulsatile or nonpulsatile?].
Because of multiple organ failure (MOF), the survival rate of patients with mechanical circulatory support has not been satisfactory, The purpose of this study is to estimate the effects of pulsatile and nonpulsatile artificial circulation on hepatic microcirculation and function. Cardiogenic shock was induced experimentally by ligating of left anterior descending branch in pigs. For the right ventricular assist device, a nonpulsatile pump (Nikkiso HPM-15) was employed. The left ventricular function was supported by either a nonpulsatile pump (Nikkiso HPM-15: NP group) or a pulsatile pump (Zeon Medical: P group). NP group was further divided into 80% support (NP-1 group) and 100% support (NP-2 group) of the control cardiac output. All groups were maintained at an equivalent mean aortic pressure of 3 hours. We measured the hepatic artery blood flow, portal vein flow and hepatic regional blood flow. For the metabolic and hepatic oxygen metabolic data, GOT, GPT, arterial blood ketone body ratio (AKBR), lactate/pirubic acid (L/P), and hyaluronic acid were evaluated. The mean aortic pressure was higher in the NP-2 group than in the other groups. The hepatic arterial blood flow was significantly higher in the P group than in the others. The AKBR and hepatic oxygen metabolism showed significant improvement in the P group in comparison with others. The regional blood flow in the liver showed improvements in the P and NP-2 groups. These findings suggested that pulsatile circulation may be beneficial for microcirculation of the liver; and the augmented nonpulsatile flow had effects similar to those of pulsatile flow in hepatic circulation.


Neuropsychological correlates of N400 anomalies in patients with schizophrenia: A preliminary report.
We investigated N400 anomalies and their relationship with neuropsychological disturbance of schizophrenia. Twelve patients with schizophrenia and 12 normal controls, matched for age, sex, education and handedness underwent both the neuropsychological test and the electrophysiological recordings employing semantic violation sentence paradigm. The patients with schizophrenia showed a reduced N400 amplitude and worse performance in the frontal lobe function test compared to healthy participants. Furthermore, statistically significant positive correlations were found between N400 amplitude and neuropsychological performances on the Stroop Task and Wisconsin Card Sorting Test in patients with schizophrenia. Our results suggest the possibility that N400 anomalies reflect the disturbed integrity of the fronto-temporal network in schizophrenia evidenced by neuropsychological deficits. In addition, we concluded that the N400 amplitude is a candidate for an endophenotype marker of schizophrenia by revealing its relationship to neuropsychological deficits.


Children and violence.
In this "information era" it can no longer be said that children are being raised in Eden or in a gentle environment of kindness and love. However rural their home, children will undoubtedly see depictions of violence on television, in the movies, or in newspapers, or hear about it on the radio or while at school or on classroom computers. All children require safety education in order to learn how to protect themselves from harm at home, at school, or in the neighborhood. This article outlines how violence may impact today's children who seek medical care.


Inflammasome-mediated Inflammation by Malassezia in human keratinocytes: A comparative analysis with different strains.
Malassezia species are associated with several common dermatologic conditions including pityriasis versicolor, seborrhoeic dermatitis, folliculitis, and atopic dermatitis and dandruff. However, its causal role remains to be established. We intended to explore the role of inflammasome activation in human keratinocytes in response to three different Malassezia species. We compared the different activation patterns of inflammasomes and the expression of pro-inflammatory cytokines and antimicrobial peptides by three different Malassezia species-M. restricta, M. globosa and M. sympodialis-in human keratinocytes. We found that different Malassezia species, especially M. restricta and M. globosa could induce nucleotide-binding oligomerisation domain, leucine-rich repeat and pyrin-domain-containing protein (NLRP)3-apoptosis-associated speck-like protein containing CARD (ASC) inflammasome activation and subsequent interleukin (IL)-1β secretion in human keratinocytes. Malassezia species variably induced thymic stromal lymphopoietin, β-defensin 2, and LL-37. IL-8 mRNA and IL-22 protein significantly increased in the M. sympodialis-treated group, and Chemokine C-C motif ligand (CCL)17 and CCL22 mRNA were increased in response to M. globosa- and M. restricta- treated keratinocytes, respectively. Our data show that various species of Malassezia promote variable inflammatory responses in keratinocytes by activating NLRP3 inflammasomes, pro-inflammatory cytokines and chemokines, and antimicrobial peptides.


The extended farm effect: The milk protein β-lactoglobulin in stable dust protects against allergies.
The allergy- and asthma-protective farm effect is mediated by numerous factors. Especially dust from cattle stables and raw cow's milk show beneficial properties, suggesting a bovine protein to be involved. As a major milk protein and member of the lipocalin family, β-lactoglobulin (BLG) binds small, hydrophobic ligands and thereby modulates the immune response. Empty BLG promotes allergy development, whereas BLG in association with ligands shows allergy-preventive as well as allergy-reducing effects in vivo and in vitro. BLG has been identified as a major protein in stable dust (therein bound to zinc) as well as in the air around cattle stables. This association with zinc favors an allergy-protective immune profile. Its immune-modulating, allergy-protective characteristics together with its presence in raw cow's milk as well as in stable dust and ambient air render BLG an essential contributor to the farm effect.


Myelin basic protein and its antibodies in the cerebrospinal fluid in experimental allergic encephalomyelitis, multiple sclerosis and other diseases.
Our studies have revealed no essential differences between EAE in monkeys and MS and other CNS diseases in humans. High concentrations of BP occur early, especially if no anti-BP antibodies are also present. Lower concentrations of BP follow and may be associated with the presence of anti-BP antibodies. In EAE these antibodies come from the relatively strong peripheral sensitization to BP and enter the CSF through a damaged blood-brain barrier; in MS they come from the relatively weak immunologic stimulation probably evoked by previous attacks of the disease. Proteolytic enzymes also enter the CSF and produce peptide fragments of BP whose differing antigenic compositions permit antibodies to some fragments to coexist with other fragments unrelated antigenically but detected as "BP" in vitro. Since oligoclonal immunoglobulins (IgG) occur more often in MS than in other diseases, one can expect to find BP, anti-BP antibodies and oligoclonal IgG more often in MS, but even the combination is not specific for MS. Consideration of the temporal and immunochemical relationships as well as of the differential diagnosis provides a basis for the understanding of the significance of BP, its antibodies and other immunoglobulins in the CSF.


Optimizing and simplifying post-traumatic amnesia testing after moderate-severe traumatic brain injury despite common confounders in routine practice.
The duration of post-traumatic amnesia (PTA) following traumatic brain injury (TBI) is a key diagnostic and outcome indicator. However, concerningly, different PTA paradigms record different PTA durations: some over-estimate, others under-estimate, PTA. Thus, a compromise is implied. The potential effect of in-hospital confounders including opioids is unknown. Three clinical groups were prospectively recruited. Group-1: in-patients with moderate-severe-TBI (MS-TBI), considered likely 'in-PTA'. Group-2: patients rehabilitating after recent MS-TBI, considered 'out-of-PTA'. Group-3: orthopaedic in-patients without TBI undergoing elective surgery. Only Groups 1&3 were taking opioids. All were administered the Westmead Post-traumatic Amnesia Scale (WPTAS) and the Galveston Orientation and Amnesia Test (GOAT). Results were obtained in n = 56 (Group-1:n = 18, Group-2:n = 13 and Group-3:n = 25). On WPTAS, Groups 1&3 scored similarly, but significantly lower than, Group-2 (χ2 = 8.2, P = 0.017). Contrariwise, on GOAT, Group-1 scored significantly lower than Groups 2&3 (χ2 = 23.99, P < 0.001): however, no patient scored GOAT <75. WPTAS showed moderate sensitivity (72%) but poor specificity (40%) in distinguishing Group-1 from Groups 2&3. Contrariwise, GOAT showed 100% specificity but 0% sensitivity. WPTAS 'day of week' and 'pictures' combined with GOAT 'transport medium to hospital', 'anterograde amnesia' and 'retrograde amnesia' maximized sensitivity (100%), specificity (85-88%), PPV (77-83%) and NPV (100%) in distinguishing Group-1 from Groups 2&3. CONCLUSIONS: Confounders including opioids likely affected WPTAS overall, but not GOAT specificity. A merger, whereby WPTAS sensitivity augmented GOAT specificity, was therefore sought. Favourable items from WPTAS (4/12) and GOAT (3/10) together optimized, and yet simplified, PTA testing; despite prevalent clinical confounders. Less, not more, 'PTA' items would benefit both patients and staff alike.


Prenatal low dosage dioxin (TCDD) exposure impairs cochlear function resulting in auditory neuropathy.
2,3,7,8-tetrachorodibenzo-p-dioxin (TCDD), a ubiquitous and persistent environmental contaminant, is a potent teratogen. Whereas developmental TCDD toxicity is mediated by the aryl hydrocarbon receptor (AhR), the normal function of the AhR is poorly understood. We tested whether dioxin exposure during a critical period of hair cell development disrupts cochlear function in three mouse strains, (C57BL6, BalbC, and CBA) that contain high affinity AhR-b alleles. C57BL/6, BalbC, and CBA dams were exposed to 500 ng/kg TCDD or olive oil (vehicle) on embryonic day 12 by gavage. Cochlear function was analyzed at 1.5 months of age by measuring 1) auditory brainstem response (ABRs) to tone pips from 5.6 to 30 kHz, and 2) distortion-product otoacoustic emissions (DPOAEs) evoked by primaries with f2 at the same frequency values. Cochlear threshold sensitivity following TCDD exposure was significantly elevated in both female and male mice in the C57BL/6 strain, carrying the Ahb-1 allele, but not significantly elevated in the BalbC or CBA strains, carrying the Ahb-2 allele. These ABR threshold deficits in mice carrying the Ahb-1 allele parallels the cleft palate incidence to higher TCDD exposures, suggesting that ABR testing could serve as a sensitive indicator of TCDD toxicity in at-risk children. Moreover, DPOAEs were not affected following TCDD exposure in any of the mouse strains, suggesting that following TCDD exposure mice with the Ahb-1 allele exhibit a mild auditory neuropathy. The causes of many auditory neuropathies are unknown, yet a developmental exposure to dioxin may be a risk factor for this condition.


Micro-Computed Tomography Analysis of the Root Canal Morphology of Palatal Roots of Maxillary First Molars.
A thorough knowledge of root canal anatomy is critical for successful root canal treatments. This study evaluated the internal anatomy of the palatal roots of maxillary first molars with micro-computed tomography (microCT). The palatal roots of extracted maxillary first molars (n = 169) were scanned with microCT to determine several anatomic parameters, including main canal classification, lateral canal occurrence and location, degree of curvature, main foramen position, apical constriction presence, diameters 1 and 2 mm from the apex and 1 mm from the foramen, minor dentin thickness in those regions, canal volume, surface area, and convexity. All canals were classified as Vertucci type I. The cross sections were oval in 61% of the canals. Lateral canals were found in 25% of the samples. The main foramen did not coincide with the root apex in 95% of the cases. Only 8% of the canals were classified as straight. Apical constriction was identified in 38% of the roots. The minor and major canal diameters and minor dentin thickness were decreased near the apex. The minor dentin thickness 1 mm from the foramen was 0.82 mm. The palatal canals exhibited a volume of 6.91 mm(3) and surface area of 55.31 mm(2) and were rod-shaped. The root canals of the palatal roots were classified as type I. However, some factors need to be considered during the treatment of these roots, including the frequent ocurrence of moderate/severe curvatures, oval-shaped cross-sections, and lateral canals, noncoincidence of the apical foramen with the root apex, and absence of apical constriction in most cases.


Compensation claims for occupational noise induced hearing loss between 1998 and 2008: yearly incidence rates and trends in older workers.
To estimate yearly incidence rates for occupational noise induced hearing loss (ONIHL) claims and to describe occupational factors in relation to age for incident cases in Victoria, Australia, between 1998 and 2008. All compensation claims lodged for deafness between 1 July 1998 and 30 June 2008 in the working population covered by the Victorian compensation scheme were analysed. Denominators were provided from 1999-2000 and incidence rates were expressed per 100,000 workers for each financial year. Overall, 81.2% of the 4,518 claims lodged were accepted. Successful claimants were predominantly males (96.5%) and claimants aged 56 to 65 years formed half the overall claims. The number of accepted claims was almost five times higher in 2007-08 than 1998-99. The highest rise was in claimants aged 56 years and above, particularly in those after retirement age. The number of claims and yearly incidence rates (IR) more than doubled over the period (240 claims and IR of 15.1 per 100,000 workers in 1999-2000 versus 669 claims and IR of 34.2 in 2007-08) with a sharp increase from 2004-05 to almost double within one single year and remained at high levels afterwards. The dramatic increase in eligible claims may reflect an increase in awareness of entitlements among workers eligible to make a successful claim. This awareness may be the result of increased opportunities for screening coinciding with changes in regulations. Older workers who worked in smaller workplaces may also be targeted by services providers as they combine occupational noise induced hearing loss (ONIHL) and presbycusis (hearing loss due to age), but this hypothesis needs further evaluation.


Unique molecular and cellular features of acute myelogenous leukemia stem cells.
It is well known in the field of acute myelogenous leukemia (AML) that many different translocations and genetic aberrancies are found with the various forms of the disease. Indeed, specific translocations are often associated with disease subtypes that manifest themselves through the accumulation of immature myeloid cells at varying stages of differentiation. Moreover, the differentiation state of myeloid blast populations has been utilized as a means of categorizing different AML subtypes (French, American, British, or FAB classification system). Thus, the notion that AML is a family of related but distinct diseases is a common view. Interestingly, however, studies in recent years that have formalized the concept of a leukemic stem cell (LSC) have also begun to define shared developmental, cellular and molecular features amongst the malignant stem cells that give rise to different AML subtypes. Moreover, some of these conserved features appear to be unique to the leukemia stem/progenitor cell population, and are not found in normal hematopoietic stem cells (HSCs). This article will summarize data emerging from the study of LSCs and suggest how distinct molecular and cellular characteristics of the LSC population may provide new opportunities for AML therapy.


Hypothenar hammer syndrome: seventeen cases with long-term follow-up.
Hypothenar hammer syndrome is a rare and often incorrectly diagnosed form of secondary Raynaud's phenomenon in workers who use their hand as a hammer. We report 17 documented cases with long-term follow-up after diagnosis made on the basis of arteriography. Clinical findings include male sex, unilaterality, sudden onset, and severe Raynaud's phenomenon. Angiography indicated that the 17 patients had either ulnar thrombosis or ulnar aneurysm; most of them also had embolic occlusions of the digital arteries. Main pathologic findings were thrombosis on the intima and fibrosis in the media. We adopted a surgical procedure consisting of resection with end-to-end reconstruction for patent aneurysms to avoid downstream emboli and more conservative treatment when the ulnar artery was thrombosed. No patient required digital amputation and all except one improved and were able to live and work normally with only a moderate disability, consisting of Raynaud's phenomenon during the cold season only.


Identification of genes showing altered DNA methylation and gene expression in the renal proximal tubular cells of rats treated with ochratoxin A for 13 weeks.
Ochratoxin A (OTA) is a mycotoxin that causes renal carcinogenicity following the induction of karyomegaly in proximal tubular cells after repeated administration to rats. Here, we performed gene profiling regarding altered DNA methylation and gene expression in the renal tubules focusing on the mechanism of OTA-induced carcinogenesis. For this purpose, OTA or 3-chloro-1,2-propanediol (3-MCPD), a renal carcinogen not inducing karyomegaly, was administered to rats for 13 weeks, and DNA methylation array and RNA sequencing analyses were performed on proximal tubular cells. Genes for which OTA altered the methylation status and gene expression level, after excluding genes showing similar expression changes by 3-MCPD, were subjected to confirmation analysis of the transcript level by real-time reverse-transcription PCR. Gene Ontology (GO)-based functional annotation analysis of validated genes revealed a cluster of hypermethylated and downregulated genes enriched under the GO term "mitochondrion," such as those associated with metabolic reprogramming in carcinogenic process (Clpx, Mrpl54, Mrps34, and Slc25a23). GO terms enriched for hypomethylated and upregulated genes included "response to arsenic-containing substance," represented by Cdkn1a involved in cell cycle arrest, and "positive regulation of IL-17 production," represented by Osm potentiating cell proliferation promotion. Other genes that did not cluster under any GO term included Lrrc14 involved in NF-κB-mediated inflammation, Gen1 linked to DNA repair, Has1 related to chromosomal aberration, and Anxa3 involved in tumor development and progression. In conclusion, a variety of genes engaged in carcinogenic processes were obtained by epigenetic gene profiling in rat renal tubular cells specific to OTA treatment for 13 weeks.


Revisiting the adequacy of infection criteria recommended in VICH GL 7 for anthelmintic effectiveness studies: Retrospective simulations.
Studies conducted to support registration or approval of veterinary anthelmintics generally follow study design recommendations provided by the VICH (International Cooperation on Harmonisation of Technical Requirements for Registration of Veterinary Medicinal Products), "Efficacy of Anthelmintics: General Requirements" (VICH GL7). For dose confirmation studies, VICH GL7 provides recommendations for determining that the control animals had an adequate infection "to permit the appropriate standards of efficacy to be met with acceptable statistical and biological certitude/confidence." In the simulation studies described in this report, we investigated the performance of one method, the statistical criterion given in Section 4.5 of VICH GL7, for evaluating the adequacy of infection in anthelmintic studies, in combination with the conventional criterion of a minimum of six adequately infected animals. We conducted numerical simulations, based on parasite data from previously conducted dose confirmation studies in dogs and cattle, to investigate how the statistical criterion impacts adequacy of infection determinations when used with the conventional criterion at various sample sizes. Simulation studies in common nematode species in both dogs and cattle indicated that under certain circumstances the statistical criterion can guard against overinterpreting the evaluation of adequacy of infection as sample size is increased. However, the statistical criterion may be overly restrictive for samples with adequate infection but containing multiple zero parasite counts and adding it to the conventional criterion does not provide any additional benefit when the sample contains no zero counts. It is important for investigators designing efficacy studies to understand the potential impact this criterion may have when establishing adequacy of infection criteria in study protocols.


Origin and yields of acetic acid in pentose-based Maillard reaction systems.
The formation of acetic acid from pentoses was studied in aqueous buffered systems (90-120 degrees C, pH 6.0-8.0) containing equimolar concentrations of 13C-labeled xylose and glycine. Acetic acid was quantified by gas chromatography-mass spectroscopy using an isotope dilution assay. Acetic acid was mainly formed from the C-1/C-2 carbon atoms of xylose (77-87%), while small amounts were also formed from the C-4/C-5 atoms of the pentose sugar (9-15%). Temperature and pH had only a small effect on the relative contribution of the sugar carbon atoms to acetic acid. These results support beta-dicarbonyl cleavage of 1-deoxypento-2,4-diulose as a major pathway leading to acetic acid in pentose-based Maillard reaction systems under food processing conditions. Acetic acid was confirmed as a major degradation product of pentoses at the early stage of the Maillard reaction, yielding 16 mol% and 28 mol% at pH 6.0 and pH 8.0, respectively.


Patients' well-being during the transition period after psychiatric hospitalization to school: insights from an intensive longitudinal assessment of patient-parent-teacher triads.
The transition period after psychiatric hospitalization back to school is accompanied by various challenges, including a substantial risk for rehospitalization. Self-efficacy and self-control, as transdiagnostic variables and important predictors of coping with school demands, should be crucial factors for successful adaptation processes as well as an overall high well-being during school reentry. The present study therefore investigates how patients' well-being develops during this period, and how it is related to patients' self-control and academic self-efficacy, as well as parents' and teachers' self-efficacy in dealing with the patient. In an intensive longitudinal design, daily ambulatory assessment measures via smartphone were collected with self-reports from the triadic perspective of 25 patients (Mage = 10.58 years), 24 parents, and 20 teachers on 50 consecutive school days, starting 2 weeks before discharge from a psychiatric day hospital (mean compliance rate: 71% for patients, 72% for parents and 43% for teachers). Patients answered daily questions between five and nine o'clock in the evening about their well-being, self-control, academic self-efficacy and about positive and negative events at school, as well as parents and teachers about their self-efficacy in dealing with the patient. Multilevel modeling revealed that on average, patients' well-being and self-control decreased during the transition period, with trends over time differing significantly between patients. While patients' academic self-efficacy did not systematically decrease over time, it did show considerable intra-individual fluctuation. Importantly, patients experienced higher well-being on days with higher self-control and academic self-efficacy as well as with higher parental self-efficacy. Daily teacher self-efficacy did not show a significant within-person relationship to daily patients' well-being. Well-being in the transition period is related to self-control and self-efficacy of patients and their parents. Thus, addressing patients' self-control and academic self-efficacy, as well as parental self-efficacy, seems promising to enhance and stabilize well-being of patients during transition after psychiatric hospitalization. Trial registration Not applicable, as no health care intervention was conducted.