Testing Prepulse Inhibition of Acoustic Startle in Rodents.
Prepulse inhibition (PPI) is a measure of sensorimotor gating which is widely used in rodents to study information processing and attention dysfunction. PPI is commonly measured in rats and mice using automated equipment. Here, we present details of a PPI testing protocol extensively used in previous studies. The protocol includes a set pulse-alone startle level and prepulse-pulse combinations with varying interval and intensity. Variations of this protocol can be used depending on the experimental aim or equipment and software version.


Shipping and Logistics Considerations for Regenerative Medicine Therapies.
Advances in regenerative medicine manufacturing continue to be a priority for achieving the full commercial potential of important breakthrough therapies. Equally important will be the establishment of distribution chains that support the transport of live cells and engineered tissues and organs resulting from these advanced biomanufacturing processes. The importance of a well-managed distribution chain for products requiring specialized handling procedures was highlighted during the COVID-19 pandemic and serves as a reminder of the critical role of logistics and distribution in the success of breakthrough therapies. This perspective article will provide insight into current practices and future considerations for creating global distribution chains that facilitate the successful deployment of regenerative medicine therapies to the vast number of patients that would benefit from them worldwide.


Multimodular Assessment of a Traumatic Bone Cyst Overlapped with Apical Periodontitis.
Traumatic bone cyst (TBC), a "pseudocyst" that usually affects long bones, is a rare lesion among cystic lesions in the jaws. The most commonly affected site is the posterior mandible. Most of the time, TBC is asymptomatic and discovered during routine radiographic examination. The treatment recommended for TBC is surgical exploration followed by curettage of the bony walls, which also serves as a diagnostic procedure. A 27-year-old Caucasian male with a noncontributory medical history was referred to our department for the endodontic evaluation of the mandibular right first and second molars, which were connected to an extensive asymptomatic osteolytic lesion. A multimodular diagnostic assessment involving CBCT imaging, ultrasound, and histopathologic examination led to a definite diagnosis of a TBC overlapping with apical periodontitis (AP). Subsequently, a multidisciplinary treatment approach was performed, including surgical excision and biopsy of the lesion, endodontic retreatment of the right mandibular first molar, and postsurgical root canal treatment of the second molar. During the follow-up period of five years, the patient was reassessed periodically once a year and showed, in the absence of signs and symptoms, progressive healing of the affected area. The present article reports a case following the CARE guidelines of a TBC combined with AP where a multimodular diagnostic assessment was performed and discusses the possible pathogenetic mechanisms involved in its generation.


Psychological Factors Are Associated with Pain at All Time Frames After Breast Cancer Surgery: A Systematic Review with Meta-Analyses.
This systematic review aimed to 1) assess associations between psychological factors and pain after breast cancer (BC) treatment and 2) determine which preoperative psychological factors predicted pain in the acute, subacute, and chronic time frames after BC surgery. A systematic review with meta-analysis. Women with early-stage BC. The Medline, EMBASE, CINAHL, and Web of Science databases were searched between 1990 and January 2019. Studies that evaluated psychological factors and pain after surgery for early-stage BC were included. Associations between psychological factors and pain, from early after surgery to >12 months after surgery, were extracted. Effect size correlations (r equivalents) were calculated and pooled by using random-effects meta-analysis models. Of 4,137 studies, 47 were included (n = 15,987 participants; 26 studies ≤12 months after surgery and 22 studies >12 months after surgery). The majority of the studies had low to moderate risk of bias. Higher preoperative anxiety and depression were weak but significant predictors of pain at all time points up to 12 months (r equivalent: 0.15-0.22). Higher preoperative pain catastrophizing and distress were also weak but significant predictors of pain during the acute (0-7 days) and chronic (3-12 months) periods (r equivalent: 0.10-0.20). For the period >12 months after surgery, weak but significant cross-sectional associations with pain were identified for anxiety, depression, pain catastrophizing, and distress (r equivalents: 0.15, 0.17, 0.25, 0.14, respectively). Significant pooled effect size correlations between psychological factors and pain were identified across all time frames. Though weak, these associations should encourage assessment of key psychological factors during preoperative screening and pain assessments at all postoperative time frames.


Puerarin Induces Hepatocellular Carcinoma Cell Apoptosis Modulated by MAPK Signaling Pathways in a Dose-dependent Manner.
Puerarin possesses a battery of therapeutic values in diverse disorders, including pro-apoptotic actions in multiple cancers. Herein, we investigated the effects of puerarin on hepatocellular carcinoma (HCC) in vitro. MTT and flow cytometry were carried out to evaluate the viability and apoptosis of SMMC-7721 HCC cells in the presence of different concentrations of puerarin. Moreover, expression levels, as well as phosphorylation status of several canonical components in mitogen-activated protein kinase (MAPK) pathways, including extracellular signal-regulated kinase 1/2 (ERK1/2), c- Jun N-terminal kinase (JNK), p38, were measured by reverse transcription and quantitative real-time polymerase chain reaction (RT-PCR) and western blot analysis at indicated time intervals. Puerarin inhibited proliferation of SMMC-7721 cells and promoted their apoptosis in a dose- and time-dependent fashion (p<0.05). Both the expression and phosphorylation levels of MAPK proteins were dramatically increased on puerarin treatment. Puerarin could be employed as a potential anti-carcinogen that exhibits pro-apoptotic effects on HCC cells, in a dose- and time-dependent manner, with emphasis on MAPK pathways whose initiation may contribute to this process.


Outbreak of Cryptocaryon irritans infection in silver pomfret Pampus argenteus cultured in China.
Silver pomfret Pampus argenteus is a major cultivated marine fish species with a high market value. In summer 2021, Cryptocaryon irritans, a ciliate parasite, infected the cultured silver pomfret in aquaculture ponds in Ningbo, Zhejiang Province, China. The symptoms of infected fish include white spots on the skin and fins, increased body surface mucus, loss of appetite, irritability, and shedding of scales. After collecting white spots from moribund fish, the 18S ribosomal RNA sequence of the pathogen on the fish skin was amplified by PCR; phylogenetic analysis showed that it was closely related to C. irritans strains from Ningde, Fujian, China. Four groups of silver pomfret were tested in an artificial infection experiment over the course of 72 h, consisting of 3 infected groups (1600, 4000, and 8000 theronts fish-1) and 1 healthy group. White spots were observed on the skin and fins of the infected fish, but not on their gills. Samples were taken from the gills, liver, kidney, and spleen of both infected and healthy fish and were compared to evaluate any significant histopathological differences. As the dose of infection increased, symptoms became more pronounced. At 72 h, mortality rates were 8.3, 50, and 66.7% for the 3 different concentrations, respectively. The median lethal concentration was calculated to be 366 theronts g-1 at 72 h, 298 theronts g-1 at 84 h, and 219 theronts g-1 at 96 h. This study emphasizes the importance of developing early diagnosis methods and appropriate prevention strategies to decrease the impact of C. irritans infection in the silver pomfret aquaculture industry.


Endoscopic management of upper tract urothelial cancer in a highly endemic area: A Taiwan nationwide collaborative study.
Endoscopic management of upper tract urothelial cancer (UTUC) is an important treatment option for low risk UTUC. Although Taiwan is an endemic area for UTUC, endoscopic treatment outcomes in Taiwan are frequently under- reported. This study retrospectively reviewed the treatment outcomes of endoscopic management for clinically localized UTUC. Patients with biopsy or washing cytology confirmed UTUC who underwent endoscopic or percutaneous management with a curative intent were retrospectively reviewed for eligibility of analysis. Those cases without pre-intervention confirmed UTUC, and metastatic or nodal disease at diagnosis were excluded. In total, 307 patients who underwent endoscopic management were reviewed and 279 cases were eligible for final analysis. With a median follow-up of 44.3 months (inter-quartile range (IQR): 23.4-76.4 months), 117 cases (46.4%) were endoscopic cured after median one session (range:1-8; IQR:1-2) of endoscopic treatment. Those endoscopic cured UTUC was associated with more small-sized tumor, more low-grade biopsied-histology, less concomitant bladder UC and less pre-operative hydronephrosis. In addition, 201(79.7%) cases among 252 cases with confirmed oncological outcome were free of UTUC at the end of follow-up and only 43 (17%) patients had a UTUC related mortality. Salvage RNU offered a better tumor free survival rate (92% vs. 77.5%) than those without salvage RNU in those UTUC refractory to endoscopic management. In multivariable analyses, pre-operative hydronephrosis was the independent risk factor for OS. Multiplicity and concomitant bladder UC were independent risk factors for DFS. We confirmed the consistent safety and efficacy of endoscopic management of clinical localized UTUC in a highly UTUC endemic area like Taiwan. Early salvage RNU is mandatory in those UTUC refractory to endoscopic management in prevention of UTUC related death.


Fe(II) complex with the octadentate btpa ligand: a DFT study on a spin-crossover system that reveals two distinct high-spin states.
Density functional theory calculations (DFT) were performed for the spin-crossover system [Fe(btpa)]2+ (btpa = N,N,N',N'-tetrakis(2-pyridylmethyl)-6,6'-bis(aminomethyl)-2,2'-bipyridine), and for the predominantly low-spin [Fe(b(bdpa))]2+ complex (in the solid state) (b(bdpa) = N,N'-bis(benzyl)-N,N'-bis(2-pyridylmethyl)-6,6'-bis(aminomethyl)-2,2'-bipyridine). The calculations confirmed that the former complex exhibits two high-spin isomers of the complexes, i.e. with C1 quasi hepta-coordinated (long-lived isomer) and C2 hexa-coordinated (short-lived isomer) structures that have been suggested previously based on time-resolved Raman and flash photolysis experiments. Application of B3LYP and B3LYP* functionals together with the CEP-31G basis yielded reasonable estimates of electronic energies (E(el) = E(el)(HS) - E(el)(LS)) for both isomers (calculated E(el) of ca. 24 and 31 kJ mol(-1) for long- and short-lived HS isomers, respectively, vs. the experimentally determined value of 27.5 kJ mol(-1)). Further calculations yielded the electronic structure of the low-spin isomer together with lowest lying singlet and triplet excited states of the [Fe(btpa)]2+ as well as the energy profile of the C2 <--> C1 isomerisation pathway for the high-spin [Fe(btpa)]2+ within the framework of the QST (quadratic synchronous transit) approach. The data obtained are discussed in relation to the observed ultrafast intersystem crossing in Fe(II) polypyridine complexes. The importance of ligand strain in relation to the destabilisation of the low-spin isomers is also discussed. In that context, calculations for a further 15 Fe(II) spin-crossover complexes of hexa-coordinating nitrogen-donor ligands have shown that the LS-HS conversion is associated with a release of ligand stress of 95 +/-16 kJ mol(-1), on average. On the basis of the calculations presented in this paper we propose that octahedral high-spin d-6 isomers are far more elastic regarding the angular distortions (equatorial and meridional strain, i.e. the declination of cis- and trans-L-M-L angle from the regular values of 90 and 180 degrees, respectively) than their low-spin counterparts.


Low-pressure hydrocephalus: indication for custom-made catheters? Technical report.
Low-pressure hydrocephalus (LPH) is characterized by ventriculomegaly with persistent low intracranial pressure (ICP). Sub-zero drainage is needed for its management and multiple solutions have been described. Our aim is to report our experience with custom-made peritoneal catheters with larger inner diameter as an alternative treatment option. We made a retrospective review of all patients diagnosed with LPH and treated with custom-made peritoneal catheters at the Virgen del Rocío Pediatric Neurosurgical Unit. Catheters were coated with antibiotic or silver. The inner diameter of ventricular catheters was 1.4 mm; peritoneal catheters were larger than usual (1.9 mm inner diameter). We identified four patients in whom five custom-made peritoneal catheters were used over a 3-year period. There were two males and the mean age was 10 years (6 months-17 years). In all patients, placement of an EVD was necessary for sub-zero drainage, with maximum negative pressure of -8 cm H20. The mean time of maintenance of EVD was 102 days (10 days-1 year). Finally, three ventriculoperitoneal (VP) valveless systems, one with antigravitation device, and one Pro-GAV VP shunt were placed, all of them with larger custom-made peritoneal catheters. After a mean follow-up period of 2.3 years (6 months-3 years), two patients are completely recovered, one patient is partially dependent for daily activities with good cognitive status, and the last one is a child who died due to his brain tumor. The custom-made peritoneal catheters with larger inner diameter could be a good option for the management of this complex pathology.


Refuting a misguided campaign against the goal of single-embryo transfer and singleton birth in assisted reproduction.
Much recent progress has been made by assisted reproductive technology (ART) professionals toward minimizing the incidence of multiple pregnancy following ART treatment. While a healthy singleton birth is widely considered to be the ideal outcome of such treatment, a vocal minority continues a campaign to advocate the benefits of multiple embryo transfer as treatment and twin pregnancy as outcome for most ART patients. Proponents of twinning argue four points: that patients prefer twins, that multiple embryo transfer maximizes success rates, that the costs per infant are lower with twins and that one twin pregnancy and birth is associated with no higher risk than two consecutive singleton pregnancies and births. We find fault with the reasoning and data behind each of these tenets. First, we respect the principle of patient autonomy to choose the number of embryos for transfer but counter that it has been shown that better patient education reduces their desire for twins. In addition, reasonable and evidentially supported limits may be placed on autonomy in exchange for public or private insurance coverage for ART treatment, and counterbalancing ethical principles to autonomy exist, especially beneficence (doing good) and non-maleficence (doing no harm). Second, comparisons between success rates following single-embryo transfer (SET) and double-embryo transfers favor double-embryo transfers only when embryo utilization is not comparable; cumulative pregnancy and birth rates that take into account utilization of cryopreserved embryos (and the additional cryopreserved embryo available with single fresh embryo transfer) consistently demonstrate no advantage to double-embryo transfer. Third, while comparisons of costs are system dependent and not easy to assess, several independent studies all suggest that short-term costs per child (through the neonatal period alone) are lower with transfers of one rather than two embryos. And, finally, abundant evidence conclusively demonstrates that the risks to both mother and especially to children are substantially greater with one twin birth compared with two singleton births. Thus, the arguments used by some to promote multiple embryo transfer and twinning are not supported by the facts. They should not detract from efforts to further promote SET and thus reduce ART-associated multiple pregnancy and its inherent risks.


HIV Lymphoma and Burkitts Lymphoma.
Despite widely available antiretroviral therapy, lymphoma remains the leading cause of death for human immunodeficiency virus (HIV)-infected persons in economically developed countries. Even a few months of drug interruptions can lead to drops in the CD4 cell count, HIV viremia, and an increased risk of lymphoma. Currently, good HIV control facilitates intensive therapies appropriate to the lymphoma, including autologous and even allogeneic hematopoietic stem cell transplantation. Nonetheless, HIV-related lymphomas have unique aspects, including pathogenetic differences driven by the presence of HIV and often coinfection with oncogenic viruses. Future therapies might exploit these differences. Lymphoma subtypes also differ in the HIV-infected population, and the disease has a higher propensity for advanced-stage, aggressive presentation and extranodal disease. Other unique aspects include the need to avoid potential interactions between antiretroviral therapy and chemotherapeutic agents and the need for HIV-specific supportive care such as infection prophylaxis. Overall, the care of these patients has progressed sufficiently that recent guidelines from the American Society of Clinical Oncology advocate the inclusion of HIV-infected patients alongside HIV-negative patients in cancer clinical trials when appropriate. This article examines HIV lymphoma and includes Burkitt lymphoma in the general population.


NK cell activity in asbestosis.
Natural killer (NK) cell activity was studied in 40 asbestos workers with chest roentgenographic signs of asbestosis and 54 sex- and age-matched healthy controls. NK cell activity in asbestosis was 32.3 +/- 18.2%, significantly higher than that in healthy controls (17.4 +/- 12.2%). We also examined the relationship between NK cell activity and the duration of exposure, and that between NK cell activity and the ILO U/C classification of radiographs. NK cell activity tended to decrease relatively in the presence of lung cancer.


On the matter of rules. Past-tense formation and its significance for cognitive neuroscience.
If neuronal models are successful, they will account for specifically human complex behaviours. Most of these behaviours can be described as governed by rules. In recent years, much effort has been spent to elucidate the neuronal basis of rules, and many researchers have focussed on modelling rules and regularities underlying language, particularly those relevant for past-tense formation. After introducing problems posed by past-tense formation, important aspects of recent controversies between connectionists and linguists concerning the nature of rules will be reviewed and analysed on the basis of elementary simulations. It is argued that modular networks with varying connection probabilities between their layers would be ideal for modelling learning and processing of past-tense formation. The motivation for postulating such networks comes from neurobiological models of language and from neuroanatomical data about cortico-cortical connectivity. Furthermore, such modular networks may explain double dissociations of regular and irregular past-tense formation in neurological patients, as reported in recent neuropsychological publications. It is concluded that past-tense formation does not pose problems to pattern associators, given that some structure is built into the network which approximates wirings in the human cortex.


Structure of DNA-porphyrin complex.
We report the 2.4 A resolution X-ray structure of a complex in which a small molecule flips a base out of a DNA helical stack. The small molecule is a metalloporphyrin, CuTMPyP4 [copper(II) meso-tetra(N-methyl-4-pyridyl)porphyrin], and the DNA is a hexamer duplex, [d(CGATCG)]2. The porphyrin system, with the copper atom near the helical axis, is located within the helical stack. The porphyrin binds by normal intercalation between the C and G of 5' TCG 3' and by extruding the C of 5' CGA 3'. The DNA forms a distorted right-handed helix with only four normal cross-strand Watson-Crick base pairs. Two pyridyl rings are located in each groove of the DNA. The complex appears to be extensively stabilized by electrostatic interactions between positively charged nitrogen atoms of the pyridyl rings and negatively charged phosphate oxygen atoms of the DNA. Favorable electrostatic interactions appear to draw the porphyrin into the duplex interior, offsetting unfavorable steric clashes between the pyridyl rings and the DNA backbone. These pyridyl-backbone clashes extend the DNA along its axis and preclude formation of van der Waals stacking contacts in the interior of the complex. Stacking contacts are the primary contributor to stability of DNA. The unusual lack of van der Waals stacking contacts in the porphyrin complex destabilizes the DNA duplex and decreases the energetic cost of local melting. Thus extrusion of a base appears to be facilitated by pyridyl-DNA steric clashes.


Bacterial formate hydrogenlyase complex.
Under anaerobic conditions, Escherichia coli can carry out a mixed-acid fermentation that ultimately produces molecular hydrogen. The enzyme directly responsible for hydrogen production is the membrane-bound formate hydrogenlyase (FHL) complex, which links formate oxidation to proton reduction and has evolutionary links to Complex I, the NADH:quinone oxidoreductase. Although the genetics, maturation, and some biochemistry of FHL are understood, the protein complex has never been isolated in an intact form to allow biochemical analysis. In this work, genetic tools are reported that allow the facile isolation of FHL in a single chromatographic step. The core complex is shown to comprise HycE (a [NiFe] hydrogenase component termed Hyd-3), FdhF (the molybdenum-dependent formate dehydrogenase-H), and three iron-sulfur proteins: HycB, HycF, and HycG. A proportion of this core complex remains associated with HycC and HycD, which are polytopic integral membrane proteins believed to anchor the core complex to the cytoplasmic side of the membrane. As isolated, the FHL complex retains formate hydrogenlyase activity in vitro. Protein film electrochemistry experiments on Hyd-3 demonstrate that it has a unique ability among [NiFe] hydrogenases to catalyze production of H2 even at high partial pressures of H2. Understanding and harnessing the activity of the FHL complex is critical to advancing future biohydrogen research efforts.


Delocalized Metal-Oxygen π-Redox Is the Origin of Anomalous Nonhysteretic Capacity in Li-Ion and Na-Ion Cathode Materials.
The anomalous capacity of Li-excess cathode materials has ignited a vigorous debate over the nature of the underlying redox mechanism, which promises to substantially increase the energy density of rechargeable batteries. Unfortunately, nearly all materials exhibiting this anomalous capacity suffer from irreversible structural changes and voltage hysteresis. Nonhysteretic excess capacity has been demonstrated in Na2Mn3O7 and Li2IrO3, making these materials key to understanding the electronic, chemical, and structural properties that are necessary to achieve reversible excess capacity. Here, we use high-fidelity random-phase-approximation (RPA) electronic structure calculations and group theory to derive the first fully consistent mechanism of nonhysteretic oxidation beyond the transition metal limit, explaining the electrochemical and structural evolution of the Na2Mn3O7 and Li2IrO3 model materials. We show that the source of anomalous nonhysteretic capacity is a network of π-bonded metal-d and O-p orbitals, whose activity is enabled by a unique resistance to transition metal migration. The π-network forms a collective, delocalized redox center. We show that the voltage, accessible capacity, and structural evolution upon oxidation are collective properties of the π-network rather than that of any local bonding environment. Our results establish the first rigorous framework linking anomalous capacity to transition metal chemistry and long-range structure, laying the groundwork for engineering materials that exhibit truly reversible capacity exceeding that of transition metal redox.


The Evaluation of the Potency of Newly Developed Oximes (K727, K733) and Trimedoxime to Counteract Acute Neurotoxic Effects of Tabun in Rats.
The ability of two newly developed oximes (K727, K733) to reduce tabun-induced acute neurotoxic signs and symptoms was evaluated and compared with currently available trimedoxime in rats. The neuroprotective effects of the oximes studied combined with atropine on Wistar rats poisoned with tabun at a lethal dose (380 µg/kg i.m.; 90% of LD50 value) were evaluated. Tabun-induced neurotoxicity was monitored by the functional observational battery consisting of 38 measurements of sensory, motor and autonomic nervous functions at 2 hours following tabun challenge. All tested oximes combined with atropine enable tabun-poisoned rats to survive till the end of experiment. Both newly developed oximes (K727, K733) combined with atropine were able to decrease tabun-induced neurotoxicity in the case of lethal poisoning although they did not eliminate all tabun-induced acute neurotoxic signs and symptoms. The ability of both novel bispyridinium oximes to decrease tabun-induced acute neurotoxicity was slightly lower than that of trimedoxime. Therefore, the newly developed oximes are not suitable for the replacement of commonly used oximes such as trimedoxime in the treatment of acute tabun poisonings.


High genetic diversity and low population structure in Porter's sunflower (Helianthus porteri).
Granite outcrops in the southeastern United States are rare and isolated habitats that support edaphically controlled communities dominated by herbaceous plants. They harbor rare and endemic species that are expected to have low genetic variability and high population structure due to small population sizes and their disjunct habitat. We test this expectation for an annual outcrop endemic, Helianthus porteri (Porter's sunflower). Contrary to expectation, H. porteri has relatively high genetic diversity (H e = 0.681) and relatively low genetic structure among the native populations (F ST = 0.077) when compared to 5 other Helianthus species (N = 288; 18 expressed sequence tag-SSR markers). These findings suggest greater gene flow than expected. The potential for gene flow is supported by the analysis of transplant populations established with propagules from a common source in 1959. One population established close to a native population (1.5 km) at the edge of the natural range is genetically similar to and shares rare alleles with the adjacent native population and is distinct from the central source population. In contrast, a transplant population established north of the native range has remained similar to the source population. The relatively high genetic diversity and low population structure of this species, combined with the long-term success of transplanted populations, bode well for its persistence as long as the habitat persists.


Mood variability predicts the course of suicidal ideation in individuals with first and second episode psychosis.
Suicide risk is high in early psychosis. Recent research has suggested that mood variability may be associated with levels of suicidal thoughts and behaviour. This has not been investigated in individuals during and following a first or second episode of non-affective psychosis. Repeated-measures data over 18 months from a large randomised controlled trial for cognitive behaviour therapy (N=309) were analysed using latent growth curve modelling, whereby both the variability and the level of depression, anxiety and guilt were entered as predictors of suicidality. The variability of depression, but not guilt and anxiety, predicted the course of suicidality even when controlling for a large range of potential confounders. The level of depression, anxiety and guilt for each participant also strongly predicted the development of suicidality. The findings support the theory that variability in depression may contribute to the formation of suicidal ideation and related behaviour. More variable depression may be harder to predict and intervene against, and therefore increase the likelihood that suicidality escalates. The levels of emotions may also be an important determinant. This has implications for the treatment and assessment of suicidality in early psychosis.


SET-CAN Fusion Gene in Acute Leukemia and Myeloid Neoplasms: Report of Three Cases and a Literature Review.
To investigate the characteristics of hematological malignancies in patients with the SET-CAN fusion gene and provide a literature review. We retrospectively analyzed the clinical data of three cases of acute leukemia and myeloid neoplasms harboring the SET-CAN fusion gene who were treated at our hospital. Their clinical manifestations, pathological results and treatment strategies were investigated. The three cases were diagnosed with T-cell acute lymphoblastic leukemia (T-ALL), acute myeloid leukemia (AML) and myeloid sarcoma (MS), respectively. Karyotype analyses identified a normal result in all three patients. Subsequently, we confirmed del(9q34) utilizing FISH analysis. Mutation of the BRAF gene was detected in case 1, while mutations in PHF6 and BCOR were detected in case 2, which have not been officially reported in patients with SET-CAN fusions. Finally, relevant literature focusing on adult patients with hematological malignancies harboring the SET-CAN fusion gene were summarized. Adult patients with the SET-CAN fusion gene were rare among cases of hematological malignancies. There was a large degree of heterogeneity between different patients. Notably, some patients remained sensitive to chemotherapy. Overall prognosis may be related to the type of disease and other cytogenetic abnormalities. Systemic cytogenetic and molecular studies are needed to make accurate diagnoses. Additional cases need to be accumulated and summarized to better understand these diseases.


Anaerobic regulation of pyruvate formate-lyase from Escherichia coli K-12.
The anaerobic regulation of the gene encoding pyruvate formate-lyase from Escherichia coli was investigated. Expression of a pfl'-'lacZ protein fusion demonstrated that the gene is subject to a 12-fold anaerobic induction which can be stimulated a further 2-fold by the addition of pyruvate to the growth medium. Construction of a strain deleted for pfl verified that either pyruvate or a metabolite of glycolysis functions as an inducer of pfl gene expression. Complete anaerobic induction required the presence of a functional fnr gene product. However, the dependence was not absolute since a two- to threefold anaerobic induction could still be observed in an fnr mutant. These results could be confirmed immunologically by analyzing the levels of pyruvate formate-lyase protein present in cells grown under various conditions. It was also shown that pfl'-'lacZ expression was partially repressed by nitrate and that this repression was mediated by the narL gene product.


CT and MR imaging of primary cardiac malignancies.
Primary cardiac malignancies are rare tumors that are difficult to diagnose clinically. Different primary cardiac malignancies may have different clinical, morphologic, and radiologic features and intracardiac locations. Angiosarcoma is the most common primary cardiac malignancy. It tends to occur in the right atrium and involve the pericardium. Because of its tendency to hemorrhage, angiosarcoma often demonstrates areas of increased signal intensity with T1-weighted sequences. Undifferentiated sarcomas typically occur in the left atrium and have variable epidemiologic and radiologic features. Rhabdomyosarcoma is the most common primary cardiac malignancy in children and is more likely than other primary cardiac sarcomas to involve the valves. Primary cardiac osteogenic sarcoma almost always occurs in the left atrium and frequently demonstrates calcification. Certain features (eg, broad base of attachment, origin at a site other than the atrial septum) help differentiate this tumor from left atrial myxoma. Leiomyosarcoma favors the left atrium and tends to invade the pulmonary veins and mitral valve. Fibrosarcoma also tends to occur in the left atrium and is often necrotic. Liposarcoma is very rare and usually manifests as a large, infiltrating mass. Foci of macroscopic fat are occasionally seen. Primary cardiac lymphoma occurs more commonly in immunocompromised patients, frequently involves the pericardium, and, unlike other primary cardiac malignancies, may respond to chemotherapy. The advent of cross-sectional imaging has allowed earlier detection of primary cardiac malignancies as well as more accurate diagnosis and characterization.


Identification of DNA binding proteins which may regulate phosphoenolpyruvate carboxykinase gene.
Transcription of phosphoenolpyruvate carboxykinase (PEPCK) gene is induced in response to cyclic AMP (cAMP) or cAMP elevating hormones. The role of transcription factors (DNA binding proteins) in the induction process has been studied. Two nuclear proteins, apparent mol. wt of 53 and 30 kDa, have been shown to bind to the 5'-flanking DNA of PEPCK gene which contains hormonal responsive elements as well as TATA box. DNA binding activity of 53 kDa protein increases by 3.5 fold in cells treated with 8-(4-chlorophenylthio)-cAMP (8-CPT-cAMP). The increased binding activity may be due to the phosphorylation of this protein by an activated cAMP-dependent protein kinase (cA kinase) in treated cells. Based on this observation, a hypothesis that 53 kDa may be specific transcription factor for PEPCK and therefore, play a major role in the regulation of this gene is proposed.


Resolution of chiasmata in oocytes requires separase-mediated proteolysis.
In yeast, resolution of chiasmata in meiosis I requires proteolytic cleavage along chromosome arms of cohesin's Rec8 subunit by separase. Since activation of separase by the anaphase-promoting complex (APC/C) is supposedly not required for meiosis I in Xenopus oocytes, it has been suggested that animal cells might resolve chiasmata by a separase-independent mechanism related to the so-called "prophase pathway" that removes cohesin from chromosome arms during mitosis. By expressing Cre recombinase from a zona pellucida promoter, we have deleted a floxed allele of separase specifically in mouse oocytes. This prevents removal of Rec8 from chromosome arms and resolution of chiasmata. It also hinders extrusion of the first polar body (PBE) and causes female sterility. mRNA encoding wild-type but not catalytically inactive separase restores chiasma resolution. Both types of mRNA restore PBE. Proteolytic activity of separase is therefore essential for Rec8's removal from chromosome arms and for chiasma resolution but not for PBE.


Monoclonal gammopathy in lymphoma.
Serum protein electrophoresis was performed in 68% of 1,682 consecutive patients with lymphoma. Of 400 patients with chronic lymphocytic leukemia and lymphocytic lymphoma, 2.3% had an IgG peak, a frequency significantly higher than that found in normal individuals of comparable age. IgM peaks occurred in 4.5% of patients with lymphomas characterized by diffuse histologic infiltration of lymph nodes. This frequency was about 100 times greater than predicted and indicated that most IgM peaks detected by electrophoresis are produced by lymphomas. Clinical responses to therapy were associated with reductions of tumor mass usually ranging from 10% to 50% of the pretreatment level. A much greater magnitude of tumor reduction is needed to improve remission duration for patients with chronic lymphocytic leukemia and lymphocytic lymphoma.


Pharmacokinetic profile that reduces nephrotoxicity of gentamicin in a perfused kidney-on-a-chip.
Nephrotoxicity is often underestimated because renal clearance in animals is higher compared to in humans. This paper aims to illustrate the potential to fill in such pharmacokinetic gaps between animals and humans using a microfluidic kidney model. As an initial demonstration, we compare nephrotoxicity of a drug, administered at the same total dosage, but using different pharmacokinetic regimens. Kidney epithelial cell, cultured under physiological shear stress conditions, are exposed to gentamicin using regimens that mimic the pharmacokinetics of bolus injection or continuous infusion in humans. The perfusion culture utilized is important both for controlling drug exposure and for providing cells with physiological shear stress (1.0 dyn cm(-2)). Compared to static cultures, perfusion culture improves epithelial barrier function. We tested two drug treatment regimens that give the same gentamycin dose over a 24 h period. In one regimen, we mimicked drug clearance profiles for human bolus injection by starting cell exposure at 19.2 mM of gentamicin and reducing the dosage level by half every 2 h over a 24 h period. In the other regimen, we continuously infused gentamicin (3 mM for 24 h). Although junctional protein immunoreactivity was decreased with both regimens, ZO-1 and occludin fluorescence decreased less with the bolus injection mimicking regimen. The bolus injection mimicking regimen also led to less cytotoxicity and allowed the epithelium to maintain low permeability, while continuous infusion led to an increase in cytotoxicity and permeability. These data show that gentamicin disrupts cell-cell junctions, increases membrane permeability, and decreases cell viability particularly with prolonged low-level exposure. Importantly a bolus injection mimicking regimen alleviates much of the nephrotoxicity compared to the continuous infused regimen. In addition to potential relevance to clinical gentamicin administration regimens, the results are important in demonstrating the general potential of using microfluidic cell culture models for pharmacokinetics and toxicity studies.


Development of an amplicon-based sequencing approach in response to the global emergence of mpox.
The 2022 multicountry mpox outbreak concurrent with the ongoing Coronavirus Disease 2019 (COVID-19) pandemic further highlighted the need for genomic surveillance and rapid pathogen whole-genome sequencing. While metagenomic sequencing approaches have been used to sequence many of the early mpox infections, these methods are resource intensive and require samples with high viral DNA concentrations. Given the atypical clinical presentation of cases associated with the outbreak and uncertainty regarding viral load across both the course of infection and anatomical body sites, there was an urgent need for a more sensitive and broadly applicable sequencing approach. Highly multiplexed amplicon-based sequencing (PrimalSeq) was initially developed for sequencing of Zika virus, and later adapted as the main sequencing approach for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Here, we used PrimalScheme to develop a primer scheme for human monkeypox virus that can be used with many sequencing and bioinformatics pipelines implemented in public health laboratories during the COVID-19 pandemic. We sequenced clinical specimens that tested presumptively positive for human monkeypox virus with amplicon-based and metagenomic sequencing approaches. We found notably higher genome coverage across the virus genome, with minimal amplicon drop-outs, in using the amplicon-based sequencing approach, particularly in higher PCR cycle threshold (Ct) (lower DNA titer) samples. Further testing demonstrated that Ct value correlated with the number of sequencing reads and influenced the percent genome coverage. To maximize genome coverage when resources are limited, we recommend selecting samples with a PCR Ct below 31 Ct and generating 1 million sequencing reads per sample. To support national and international public health genomic surveillance efforts, we sent out primer pool aliquots to 10 laboratories across the United States, United Kingdom, Brazil, and Portugal. These public health laboratories successfully implemented the human monkeypox virus primer scheme in various amplicon sequencing workflows and with different sample types across a range of Ct values. Thus, we show that amplicon-based sequencing can provide a rapidly deployable, cost-effective, and flexible approach to pathogen whole-genome sequencing in response to newly emerging pathogens. Importantly, through the implementation of our primer scheme into existing SARS-CoV-2 workflows and across a range of sample types and sequencing platforms, we further demonstrate the potential of this approach for rapid outbreak response.


In Vitro Evaluation of Direct and Indirect Effects of Sonic and Ultrasonic Instrumentations on the Shear Bond Strength of Orthodontic Brackets.
Sonic and ultrasonic instrumentations generate vibrations that may influence debonding characteristics. The objective of this in vitro study was to assess the direct and indirect effects of sonic and ultrasonic periodontal instrumentations on the shear bond strength (SBS) and the adhesive remnant index (ARI) scores of metallic orthodontic brackets. Metallic brackets were bonded to 75 extracted mandibular central incisors that were embedded in acrylic resin. Instrumentations around the bracket base performed with ultrasonic (UltrasonicB group, n=15) and sonic (SonicB group, n=15) scalers were used to evaluate the direct effects on the SBS of brackets. Lingual surface instrumentations with ultrasonic (UltrasonicL group, n=15) and sonic (SonicL group, n=15) scalers were performed to assess the indirect effects. The control group (n=15) did not have any treatment. Instrumentations were performed for 30 s with 0° scaler tip angulations with settings recommended by manufacturers. The SBS of the brackets tested with a universal testing machine and ARI scores were recorded. Data were analyzed by Kruskal-Wallis and Mann-Whitney U tests. The mean SBS of the control group was significantly higher than that of the UltrasonicB and SonicB groups (p=0.008). The UltrasonicL and SonicL group instrumentations also decreased the SBS, although the difference was statistically insignificant. UltrasonicB instrumentations caused significantly higher frequency of ARI scores than the control group. The decrease of the SBS of metallic brackets indicates the influence of ultrasonic and sonic instrumentations on the breakage behavior at the bracket-resin interface. Instrumentations around the bracket base should be conducted with caution to decrease the bond failure risk of metallic brackets.


Adipogenesis in thyroid eye disease.
Adipogenesis contributes to the pathogenesis of thyroid eye disease (TED). Thyrotropin receptor (TSHR) transcripts are present in orbital fat. This study was conducted to determine whether they are expressed as functional protein, and if so, whether this is restricted to TED orbits or to a particular stage in adipocyte differentiation. Samples of fat were obtained from 18 TED-affected orbits and 4 normal orbits, and 9 were obtained from nonorbital locations. Frozen sections were examined by immunocytochemistry using monoclonal antibodies specific for the human TSHR. Samples were disaggregated and the preadipocytes separated from the mature by differential centrifugation and cultured in serum-free or DM and examined for morphologic changes, oil red O and TSHR staining, and TSH-induced cyclic adenosine monophosphate (cAMP) production. Marked immunoreactivity was observed in frozen sections from all three TED samples and faint staining in both normal orbital fat samples. In vitro, 1% to 5% of preadipocytes displayed TSHR immunoreactivity in five of six TED and two of three normal orbital samples and in three of five nonorbital samples. Differentiation, was induced in all 14 orbital samples. Three of four nonorbital samples contained occasional differentiated cells. Fifty percent to 70% of differentiating cells demonstrated receptor immunoreactivity. Two of three TED and four of four nonorbital preadipocytes in DM and/or mature adipocytes displayed a TSH-mediated increase in cAMP. The results indicate that orbital fat TSHR transcripts are expressed as protein, which can be functional. This is not aberrant in TED orbits, although expression may be upregulated. The majority of preadipocytes undergoing differentiation express the receptor, indicating a key role for this population in one mechanism for increasing orbital volume.


Effective tumor therapy with plasmid-encoded cytokines combined with in vivo electroporation.
Plasmids may have unique advantages as a gene delivery system. However, a major obstacle is the low in vivo transduction efficiency. In this study, an electroporation-based gene transduction approach was taken to study the effect of interleukin (IL)-2 or IL-12 gene transduction on the growth of experimental murine tumors. Significant intratumoral gene transduction was achieved by electroporation of tumors that had been injected with naked plasmids encoding reporter genes and cytokine genes (IL-2 and IL-12) under the control of a constitutive cytomegalovirus promoter. In addition, significant tumor growth delay could be achieved in a murine melanoma line B16.F10 with the cytokine genes. Most importantly, systemic transgene levels were negligible when compared with intratumoral adenovirus-mediated IL-12 gene delivery, which leads to significantly higher systemic cytokine levels. Therefore, naked plasmid- and in vivo electroporation-mediated cancer gene therapy may be therapeutically efficacious while maintaining low systemic toxicity.


[Ultrasound anatomy of masticatory muscles. (Experimental and clinical study)].
To describe the ultrasound anatomy of the masticatory muscles based on a comparison of the results of macroscopic and ultrasound studies of the masticatory muscles in vitro. In the experimental part, a macropreparation of the masticatory muscle was studied; in a clinical example, an ultrasound of the masticatory muscles was performed on a patient with normal occlusion using the author's methods for analyzing echograms. The ultrasound anatomy of the masticatory muscles is described based on a comparison of data from the study of a macroscopic specimen and ultrasound images of the masticatory muscle of an experimental animal. Using a clinical example of a patient with normal occlusion, the results demonstrate the usage of the authors' developments in describing ultrasound images of the masticatory muscles. The study resulted in an algorithm for describing ultrasound images of masticatory muscles, including methods for qualitative and quantitative assessment of ultrasound images using proprietary developments involving elements of artificial intelligence technologies.


Effect of meat enhancement solutions with hydroxypropyl methylcellulose and konjac flour on texture and quality attributes of pale, soft, and exudative pork.
The objective of this study was to determine whether addition of hydrocolloids, buffer ingredients, salt, and sodium phosphate improve the color, texture, and pH of normal and pale, soft, and exudative (PSE) meat. Specific solutions include potassium bicarbonate (KHCO), ammonium bicarbonate (NHHCO), hydroxypropyl methylcellulose (HPMC), and konjac flour (KF). Three studies were carried out. First, the stability and viscoelastic properties of the different solutions was determined. Second, fresh normal (pH 5.6-5.9) and PSE (pHu < 5.4) Longissimus dorsi muscles were ground and assigned a treatment solution: control (fresh normal), water, NaCl/SP, KHCO, NHHCO, KF, HPMC, KHCO/KF, KHCO/KF/NaCl/SP (sodium phosphate), NHHCO/HPMC, and NHHCO/HPMC/NaCl/SP. Third, treatment solutions with suitable viscoelastic properties and stability, were added, homogenized, stuffed into tubes, and cooked to an internal temperature of 75C to form gels, and torsion and texture profile analysis performed. The PSE treated samples containing bicarbonate/hydrocolloid treatments had similar hardness-first bite, hardness-second bite, and gumminess values to the normal-control samples while the NHHCO treated samples had air pockets in the cooked gel. Third, consumers (n = 92) evaluated control and PSE beef/pork frankfurters produced with the following treatments: KHCO, KHCO/HPMC, and KHCO/KF. Treatment had no effect (p > .05) on overall like/dislike of flavor, but PSE frankfurters were preferred (p < .05) to the controls. The use of KHCO with hydrocolloids, salt and SP improved the color, pH and texture properties of PSE ground pork; further research to examine the effectiveness in whole PSE pork muscle systems is needed. Meat processors could use KHCO with HPMC or KF as ingredients to improve color, texture, and pH of PSE meat. The reduction of variation between PSE and normal pork muscle would improve pork quality and add value to PSE meat products.


The language of pain.
In a well-known advertising campaign, respondents asked to spell "relief" invariably spelled the name of a popular antacid. Three decades ago, an American nurse working in Sweden found less unanimity, however, when it came to the spelling of "pain." Intrigued by the clear distinction that her patients made between pain and ache, Fannie Gaston-Johansson wanted to know more. Did the same differentiation transcend cultural and linguistic divides? Was it possible to develop a simple, word-based tool to reliably quantify pain? Her search for these and other answers would result in outstanding scholarship, a patented invention and history-making achievement.


Effective DBS treatment improves neural information transmission of patients with disorders of consciousness: an fNIRS study.
Deep Brain Stimulation (DBS) is a potential treatment that promotes the recovery of patients with disorders of consciousness (DOC). This study quantified the changes in consciousness and the neuromodulation effect of DBS on patients with DOC. Eleven patients were recruited for this study which consists of three conditions: 'Pre' (two days before DBS surgery), 'Post-On' (one month after surgery with stimulation), and 'Post-Off' (one month after surgery without stimulation). Functional near-infrared spectroscopy (fNIRS) was recorded from the frontal lobe, parietal lobe, and occipital lobe of patients during the experiment of auditory stimuli paradigm, in parallel with the Coma Recovery Scale-Revised (CRS-R) assessment. The brain hemodynamic states were defined and state transition acceleration was taken to quantify the information transmission strength of the brain network. Linear regression analysis was conducted between the changes in regional and global indicators and the changes in the CRS-R index. Significant correlation was observed between the changes in the global transition acceleration indicator and the changes in the CRS-R index (slope=55.910,p< 0.001,R2= 0.732). For the regional indicators, similar correlations were found between the changes in the frontal lobe and parietal lobe indicators and the changes in the CRS-R index (slope=46.612,p< 0.01, R2 = 0.694; slope=47.491,p< 0.01,R2= 0.676). Our study suggests that fNIRS-based brain hemodynamics transition analysis can signify the neuromodulation effect of DBS treatment on patients with DOC, and the transition acceleration indicator is a promising brain functional marker for DOC.


Gene transfer of kringle 5 of plasminogen by electroporation inhibits corneal neovascularization.
To test the efficacy of naked plasmid that expresses human kringle 5 of plasminogen (K5) in suppressing experimental corneal neovascularization in a rat model. A eukaryotic expression plasmid encoding human K5 (pSecK5) was constructed. COS cells were transiently transfected with pSecK5 using a lipid-based transfection reagent. K5 secretion was confirmed by Western blot analysis. The effect of the secreted K5 on the proliferation of human umbilical vein endothelial cells (HUVECs) was investigated colorimetrically. Forty-three Sprague-Dawley rats were used for a corneal neovascularization suppression experiment. Corneal injury was induced by placing a disk of filter paper (immersed in 1 mol/l NaOH, 3.0 mm in diameter) on the corneal surface for 2 min. The cornea was immediately washed with saline. pSecK5 and empty plasmids were injected subconjunctivally, and square-wave electric pulses were immediately applied to the eyes. The expression of K5 was analyzed by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. The extent of corneal neovascularization was evaluated by scores. The constructed plasmid could express itself in COS cells. Conditioned medium from K5-transfected COS cells apparently inhibited HUVEC proliferation, compared with conditioned medium from COS cells transfected with empty plasmid or nontransfected cells. RT-PCR and immunohistochemistry confirmed the expression of K5 in the conjunctiva and cornea. Corneal neovascularization was significantly suppressed by K5 gene transfer in rats' eyes. In a rat model, K5 gene transfer by subconjunctival injection and electroporation can effectively inhibit corneal neovascularization induced by an alkali burn.


Minimum anesthetic concentration of isoflurane in captive thick-billed parrots (Rhynchopsitta pachyrhyncha).
To determine the minimum anesthetic concentration (MAC) of isoflurane in thick-billed parrots (Rhynchopsitta pachyrhyncha). 15 healthy thick-billed parrots. Anesthesia was induced and maintained with isoflurane in oxygen. In the first bird that was anesthetized, end-tidal isoflurane concentration was maintained at 1.0% for 15 minutes. After this period of anesthetic equilibration, an end-tidal gas sample was obtained for verification of isoflurane concentration. A toe was pinched to determine the bird's response to pain, and the bird was then allowed to recover from aesthesia. To determine MAC, a so-called up-and-down approach was subsequently used in all 15 birds. Compared with the isoflurane concentration used for MAC determination in the first bird, maintenance isoflurane concentration for the second bird was increased by approximately 10% if the first bird reacted and decreased by approximately 10% if the first bird did not react to a toe pinch. These steps were then followed until all 15 birds had been anesthetized. Crossover events occurred when birds in sequence had discordant results (ie, 1 reactor and 1 nonreactor). The MAC was defined as the mean of the isoflurane concentrations measured during these crossover events. Mean MAC of isoflurane in thick-billed parrots was estimated to be 1.07% (95% confidence interval, 0.97% to 1.16%). Isoflurane MAC appears to be lower in thick-billed parrots than the MAC determined for other bird species. Determination of the species-specific requirements of thick-billed parrots should allow isoflurane anesthesia to be performed more safely in this endangered species.


miRNA-30c can be used as a target in the diagnosis and treatment of osteosarcoma.
Osteosarcoma is a highly malignant osseous sarcoma with poor prognosis. Previous studies indicated that miRNA-30c may play an important role in the development of osteosarcoma, but its mechanism is not yet clear. The current research was carried out to explore the potential applications of miR-30c in the diagnosis and treatment of osteosarcoma. Real-time PCR and in situ hybridization were used to test the correlation between miR-30c and the onset of osteosarcoma. In vitro transfection of miR-30 mimic was used to test the effect of miR-30c on the development of osteosarcoma. Cell Counting Kit-8, formation of Petri dish clones, in vivo formation of tumor, flow cytometry tests and Transwell analysis were used to assess the effect of miR-330c on the metastatic potential and invasiveness of osteosarcoma. Reverse transcriptase-PCR analysis and in situ hybridization tests revealed that the expression of miR-30c was lower in the osteosarcoma tissue than in normal bone tissue (P<0.05). Low expression of miR-30c was associated with advanced osteosarcoma staging and low cellular differentiation. Multivariate analysis revealed that lower expression of miR-30c was associated with shorter survival of patients (P<0.01). U2OS cell growth was significantly inhibited when transfected with miR-30c mimic. Flow cytometry analysis revealed that overexpression of miR-30c could induce apoptosis of osteosarcoma cells. In vitro Petri dish cloning experiment showed that overexpression of miR-30c reduced the cloning ratio of U2OS cells from 21% to 7%. At the same time, overexpression of miR-30c inhibited the formation of sarcoma in nude mice. Transwell experiments indicated that overexpression of miR-30c could reduce the invasiveness of U2OS cells. Low expression of miR-30c was associated with high probability of onset and aggressiveness of osteosarcoma and shorter patient survival. Upregulation of miR-30c could downregulate the invasiveness of osteosarcoma. Therefore, miR-30 can be used in the development of future diagnostic and therapeutic techniques.


Enhancement of rAAV2-mediated transgene expression in retina cells in vitro and in vivo by coadministration of low-dose chemotherapeutic drugs.
Recombinant adeno-associated viral vector serotype 2 (rAAV2) has been used with success to deliver retina-targeted gene therapeutics in retinal degeneration. However, one of the major limitations of this approach is the vector's low transduction efficiency. This study is designed to increase AAV2 transduction efficiency in vitro and in vivo. Green fluorescence protein (GFP) or luciferase reporter gene-carried rAAV2 vectors were applied to cultured human RPE cells (ARPE-19) or animal eyes with or without chemotherapeutic agents. GFP transduction efficiency was evaluated by image, flow cytometry analysis, and Western blot. The ciliary neurotrophic factor (rAAV2-CNTF)-carried AAV2 vector was coinjected to subretinal space with or without chemotherapeutic agent. The therapeutic efficacy was evaluated by counting numbers of remaining photoreceptors in retina sections of treated or untreated eyes. Coadministration of 0.1 μg/mL doxorubicin (DXR), 0.14 μg/mL cytarabine (Ara-C), 1 μg/mL etoposide (VP-16), or 20 μg/mL cisplatin (DDP) significantly increased rAAV2-mediated GFP and/or luciferase expression in cultured hRPE cells without any detectable toxicity. Pretreatment with DXR for 24 h prior to infection was most effective in enhancing rAAV2 transgene expression in hRPE cells. In addition, subretinal coinjection of rAAV2-CMV-ciliary neurotrophic factor (rAAV2-CNTF) and DXR into the eyes of rats with inherited retinal degeneration resulted in an approximately 2-fold increase in photoreceptor layer thickness and cellular density of the outer nuclear layer (ONL) compared to rAAV2-CNTF alone, reflecting a pronounced protection effect mediated by the enhanced expression of CNTF. The method described here to improve rAAV2-based gene delivery is simple and feasible without any detectable toxicity. This strategy might be therapeutically exploited in the gene therapy of degenerative retinal diseases.


Central command generated prior to arbitrary motor execution induces muscle vasodilatation at the beginning of dynamic exercise.
The purpose of this study was to examine the role of central command, generated prior to arbitrary motor execution, in cardiovascular and muscle blood flow regulation during exercise. Thirty two subjects performed 30 s of two-legged cycling or 1 min of one-legged cycling (66 ± 4% and 35% of the maximal exercise intensity, respectively), which was started arbitrarily or abruptly by a verbal cue (arbitrary vs. cued start). We measured the cardiovascular variables during both exercises and the relative changes in oxygenated-hemoglobin concentration (Oxy-Hb) of noncontracting vastus lateralis muscles as index of tissue blood flow and femoral blood flow to nonexercising leg during one-legged cycling. Two-legged cycling with arbitrary start caused a decrease in total peripheral resistance (TPR), which was smaller during the exercise with cued start. The greater reduction of TPR with arbitrary start was also recognized at the beginning of one-legged cycling. Oxy-Hb of noncontracting muscle increased by 3.6 ± 1% (P < 0.05) during one-legged cycling with arbitrary start, whereas such increase in Oxy-Hb was absent with cued start. The increases in femoral blood flow and vascular conductance of nonexercising leg were evident (P < 0.05) at 10 s from the onset of one-legged cycling with arbitrary start, whereas those were smaller or absent with cued start. It is likely that when voluntary exercise is started arbitrarily, central command is generated prior to motor execution and then contributes to muscle vasodilatation at the beginning of exercise. Such centrally induced muscle vasodilatation may be weakened and/or masked in the case of exercise with cued start.


Decision making under time pressure: an independent test of sequential sampling models.
Choice probability and choice response time data from a risk-taking decision-making task were compared with predictions made by a sequential sampling model. The behavioral data, consistent with the model, showed that participants were less likely to take an action as risk levels increased, and that time pressure did not have a uniform effect on choice probability. Under time pressure, participants were more conservative at the lower risk levels but were more prone to take risks at the higher levels of risk. This crossover interaction reflected a reduction of the threshold within a single decision strategy rather than a switching of decision strategies. Response time data, as predicted by the model, showed that participants took more time to make decisions at the moderate risk levels and that time pressure reduced response time across all risk levels, but particularly at the those risk levels that took longer time with no pressure. Finally, response time data were used to rule out the hypothesis that time pressure effects could be explained by a fast-guess strategy.


Structural anisotropy and internal magnetic fields in trabecular bone: coupling solution and solid dipolar interactions.
We investigate the use of intermolecular multiple-quantum coherence to probe structural anisotropy in trabecular bone. Despite the low volume fraction of bone, the bone-water interface produces internal magnetic field gradients which modulate the dipolar field, depending on sample orientation, choice of dipolar correlation length, correlation gradient direction, and evolution time. For this system, the probing of internal magnetic field gradients in the liquid phase permits indirect measurements of the solid phase dipolar field. Our results suggest that measurements of volume-averaged signal intensity as a function of gradient strength and three orthogonal directions could be used to non-invasively measure the orientation of structures inside a sample or their degree of anisotropy. The system is modeled as having two phases, solid and liquid (bone and water), which differ in their magnetization density and magnetic susceptibility. A simple calculation using a priori knowledge of the material geometry and distribution of internal magnetic fields verifies the experimental measurements as a function of gradient strength, direction, and sample orientation.


Brief Report: Dapivirine Vaginal Ring Use Does Not Diminish the Effectiveness of Hormonal Contraception.
To evaluate the potential for a clinically relevant drug-drug interaction with concomitant use of a dapivirine vaginal ring, a novel antiretroviral-based HIV-1 prevention strategy, and hormonal contraception by examining contraceptive efficacies with and without dapivirine ring use. A secondary analysis of women participating in MTN-020/ASPIRE, a randomized, double-blind, placebo-controlled trial of the dapivirine vaginal ring for HIV-1 prevention. Use of a highly effective method of contraception was an eligibility criterion for study participation. Urine pregnancy tests were performed monthly. Pregnancy incidence by arm was calculated separately for each hormonal contraceptive method and compared using an Andersen-Gill proportional hazards model stratified by site and censored at HIV-1 infection. Of 2629 women enrolled, 2310 women returned for follow-up and reported using a hormonal contraceptive method at any point during study participation (1139 in the dapivirine arm and 1171 in the placebo arm). Pregnancy incidence in the dapivirine arm versus placebo among women using injectable depot medroxyprogesterone acetate was 0.43% vs. 0.54%, among women using injectable norethisterone enanthate was 1.15% vs. 0%, among women using hormonal implants was 0.22% vs. 0.69%, and among women using oral contraceptive pills was 32.26% vs. 28.01%. Pregnancy incidence did not differ by study arm for any of the hormonal contraceptive methods. Use of the dapivirine ring does not reduce the effectiveness of hormonal contraceptives for pregnancy prevention. Oral contraceptive pill use was associated with high pregnancy incidence, potentially because of poor pill adherence. Injectable and implantable methods were highly effective in preventing pregnancy.


Residual block fully connected DCNN with categorical generalized focal dice loss and its application to Alzheimer's disease severity detection.
Alzheimer's disease (AD) is a disease that manifests itself with a deterioration in all mental activities, daily activities, and behaviors, especially memory, due to the constantly increasing damage to some parts of the brain as people age. Detecting AD at an early stage is a significant challenge. Various diagnostic devices are used to diagnose AD. Magnetic Resonance Images (MRI) devices are widely used to analyze and classify the stages of AD. However, the time-consuming process of recording the affected areas of the brain in the images obtained from these devices is another challenge. Therefore, conventional techniques cannot detect the early stage of AD. In this study, we proposed a deep learning model supported by a fusion loss model that includes fully connected layers and residual blocks to solve the above-mentioned challenges. The proposed model has been trained and tested on the publicly available T1-weighted MRI-based KAGGLE dataset. Data augmentation techniques were used after various preliminary operations were applied to the data set. The proposed model effectively classified four AD classes in the KAGGLE dataset. The proposed model reached the test accuracy of 0.973 in binary classification and 0.982 in multi-class classification thanks to experimental studies and provided a superior classification performance than other studies in the literature. The proposed method can be used online to detect AD and has the feature of a system that will help doctors in the decision-making process.


Obesity and endothelial dysfunction.
Obesity is becoming more prevalent in the developed world because of the abundance of food and the decrease of physical activity. Obesity is a risk factor for a host of diseases from arthritis to cardiovascular disease. The precise mechanisms by which obesity promotes cardiovascular disease are not well understood but are likely to include metabolic and inflammatory responses to the increased amount of stored fat. The endothelium plays a pivotal role in maintaining vascular health. Impaired endothelial function is an independent predictor of cardiovascular disease. Most studies of vascular function in obese subjects have demonstrated impaired endothelial function. This impairment of endothelial function becomes obvious early on, long before any vascular abnormalities become clinically relevant and detectable. Better understanding of the mediators of obesity-induced endothelial dysfunction may lead to the identification of new targets for interventions that may prevent or postpone the development of obesity-related cardiovascular disease.


The neurotransmitter ATP triggers Ca2+ responses promoting coordination of pancreatic islet oscillations.
Pulsatile insulin release into the portal vein is critically dependent on entrainment of the islets in the pancreas into a common oscillatory phase. Because the pulses reflect periodic variations of the cytoplasmic Ca concentration ([Ca]i), we studied whether the neurotransmitters adenosine triphosphate (ATP) and acetylcholine promote synchronization of [Ca]i oscillations between islets lacking contact. Medium-sized and small mouse islets and cell aggregates were used for measuring [Ca]i with the indicator fura-2. Exposure to acetylcholine resulted in an initial [Ca]i peak followed by disappearance of the [Ca]i oscillations induced by 11-mmol/L glucose. The effect of ATP was often restricted to an elusive [Ca]i peak. The incidence of distinct [Ca]i responses to ATP increased under conditions (accelerated superfusion, small islets, or cell aggregates) intended to counteract purinoceptor desensitization owing to intercellular accumulation of ATP. Attempts to imitate neural activity by brief (15 seconds) exposure to ATP or acetylcholine resulted in temporary synchronization of the glucose-induced [Ca]i oscillations between islets lacking contact. The data support the idea that purinergic signaling has a key role for coordinating the oscillatory activity of the islets in the pancreas, reinforcing previous arguments for the involvement of nonadrenergic, noncholinergic neurons.


Dependence of groundnut rosette virus on its satellite RNA as well as on groundnut rosette assistor luteovirus for transmission by Aphis craccivora.
Transmission of groundnut rosette virus (GRV) by Aphis craccivora is known to depend on the additional presence in the source plants of a luteovirus, groundnut rosette assistor virus (GRAV). Naturally occurring isolates of GRV contain a satellite RNA which is the main cause of rosette symptoms in groundnut, different variants of the satellite being responsible for the green and chlorotic forms of rosette. In extensive glasshouse tests, GRAV-dependent transmission of GRV by A. craccivora occurred only from groundnut plants infected with satellite-containing isolates of GRV. This was true whether the GRV isolates were from groundnut plants from Nigeria or Malawi with either the green or chlorotic forms of rosette and whether they contained homologous or heterologous satellites. Aphid transmission of GRV therefore depends not only on the presence of GRAV but also on that of the GRV satellite RNA. This probably explains why satellite-free isolates of GRV have not been found in nature. This is the first report of satellite RNA mediating aphid transmission of a plant virus.


Co-liposomes comprising a lipidated multivalent RGD-peptide and a cationic gemini cholesterol induce selective gene transfection in αvβ3 and αvβ5 integrin receptor-rich cancer cells.
The αvβ3 and αvβ5 integrins, transmembrane glycoprotein receptors, are over-expressed in numerous tumors and in endothelial cells that constitute tumor blood vessels. As this protein selectively binds to the Arg-Gly-Asp (RGD) sequence containing peptides, it is an attractive way to target tumors. Herein we have developed novel formulations for integrin mediated selective gene delivery. These formulations are composed of a novel palmitoylated tetrameric RGD containing scaffold (named RAFT-RGD), cationic gemini cholesterol (GL5) and a natural helper lipid 1,2-dioleoyl-l-α-glycero-3-phosphatidylethanolamine (DOPE). We have optimized a co-liposomal formulation to introduce the multivalent RGD-containing macromolecule in GL5: DOPE (GL5D) mixture to produce GL5D-RGD. We have unambiguously shown the selectivity of these formulations towards cancer cells that over express αvβ3 and αvβ5 integrins. Two reporter plasmids, pEGFP-C3 and PGL-3, were employed for the transfection experiments and it was shown that GL5D-RGD liposomes increased exclusively the transfection in αvβ3 and αvβ5-overexpressing HeLa cells.


A new troodontid dinosaur from China with avian-like sleeping posture.
Discovering evidence of behaviour in fossilized vertebrates is rare. Even rarer is evidence of behaviour in non-avialan dinosaurs that directly relates to stereotypical behaviour seen in extant birds (avians) and not previously predicted in non-avialan dinosaurs. Here we report the discovery of a new troodontid taxon from the Early Cretaceous Yixian Formation of western Liaoning, China. Numerous other three-dimensionally preserved vertebrate fossils have been recovered recently at this locality, including some specimens preserving behavioural information. The new troodontid preserves several features that have been implicated in avialan origins. Notably, the specimen is preserved in the stereotypical sleeping or resting posture found in extant Aves. Evidence of this behaviour outside of the crown group Aves further demonstrates that many bird features occurred early in dinosaurian evolution.


Arterial complications following cardiac catheterization in children less than 10 kg.
We sought to determine if a higher dose of heparin would reduce arterial complications in patients weighing 10 kg or less undergoing cardiac catheterization to investigate congenital heart disease. Sixty patients were given either 100 (group A) or 150 (group B) IU x kg(-1) of heparin in a double-blinded randomized manner. Initial arterial access was established using a 4F cannula in all patients. Mean activated clotting time measured 20 minutes following heparin administration was significantly lower in group A than in group B (199 versus 251 seconds). Only 3 out of 60 patients (5%) required treatment for loss of femoral pulse. The age, weight, activated clotting time, length of catheterization procedure, time taken to establish arterial access, and the duration of arterial cannulation were comparable between the groups. Weight under 4 kg, age under 1 month, and cannula size larger than 4F were identified as independent risk factors for the development of arterial complications. Arterial access using a 4F cannula is a safe procedure in children weighing 10 kg or less. The incidence of significant arterial complications is low, and they do not appear to be preventable by a higher dose of heparin.


Discovery of flavone-derivatives as the new skeleton of transient receptor potential vanilloid 3 channel antagonists.
Transient receptor potential vanilloid 3 (TRPV3) channel is a temperature-sensitive and Ca2+-permeable nonselective cation channel, which is abundantly expressed in skin keratinocyte and plays an important role in skin homeostasis and repair. However, only a few TRPV3 inhibitors were reported. Few selective and potent modulators of the TRPV3 channel have hindered the progress of the investigation and clinical application. TRPV3 channel research still faces challenges and requires the new inhibitors. Flavonoids are a kind of natural compounds with various biological and pharmacological activities including anti-inflammatory and anti allergic effects, which is associated with some physiological effects mediated by TRPV3 channel. Herein, our group designed and synthesized a range of flavone derivatives, and investigated their inhibitory properties on the human TRPV3 channel by electrophysiology technique. Then, we identified a new potent TRPV3 antagonist 2d with IC50 of 0.62 μM. It also showed good selectivity on TRPV1, TRPV4, TRPA1 and TRPM8.


San fernando earthquake of 9 february 1971: pattern of faulting.
Mapping of the surface breaks that resulted from the San Fernando earthquake of 9 February 1971 reveals that the pattern of faulting was highly complex; it consisted of a number of segments that produced ground displacements and acceleration throughout the entire northern end of the San Fernando Valley. Instead of occurring on the frontal fault zone, as might have been expected, the faulting occurred on the valley side of the frontal fault system, which separates the crystalline rocks of the San Gabriel Mountains from the Tertiary sediments of the San Fernando Valley. However, the new fault system does, in many cases, follow breaks in slope and subtle escarpments that suggest faulting along these zones in the recent geologic past.


Tissue-specific silencing of homoeologs in natural populations of the recent allopolyploid Tragopogon mirus.
Recent years have seen rapid advances in our knowledge of the transcriptomic consequences of allopolyploidy, primarily through the study of polyploid crops and model systems. However, few studies have distinguished between homoeologs and between tissues, and still fewer have examined young natural allopolyploid populations of independent origin, whose parental species are still present in the same location. Here, we examined the expression of 13 homoeolog pairs in seven tissues of 10 plants of allotetraploid Tragopogon mirus from two natural populations formed by independent polyploidizations between Tragopogon dubius and Tragopogon porrifolius c. 40 generations ago. We compare these with patterns of expression in the diploid parental species from the same locality. Of the 910 assays in T. mirus, 576 (63%) showed expression of both homoeologs, 63 (7%) showed no expression of either homoeolog, 186 (20%) showed nonexpression of one homoeolog across all tissues of a plant, and 72 (8%) showed non-expression of a homoeolog in a particular tissue within a plant. We found two cases of reciprocal tissue-specific expression between homoeologs, potentially indicative of subfunctionalization. Our study shows that tissue-specific silencing, and even apparent subfunctionalization, can arise rapidly in the early generations of natural allopolyploidy.


Expression of the large T protein of polyoma virus promotes the establishment in culture of "normal" rodent fibroblast cell lines.
Transfer into mouse and rat embryo fibroblasts in primary culture of cloned polyoma virus genes encoding only the large T protein led to the establishment of flat colonies in sparse subcultures at a frequency equal to that of transformation by wild-type virus. Cell lines could be derived from such colonies and maintained in culture for large numbers of generations without entering crisis. They exhibited a normal phenotype, by the criteria of growth on plastic to a low saturation density and of anchorage dependency. However, they required a lower serum concentration for growth than spontaneously established 3T3 cells. Similar results were obtained after transfer of recombinant DNA molecules encoding only the amino-terminal 40% of the large T protein, suggesting that this "immortalization" function corresponds to the activity of an amino-terminal domain of the protein. Immunoprecipitation analysis of T antigens in cell lines established after transfer of the full-size and of the truncated large T genes demonstrated the expression of the full-size large T protein and of a Mr 40,000 antigen expressed from the amino-terminal part of the gene, respectively. After transfer of a "large T only" plasmid that carries a tsa mutation, cell lines were established at 33 degrees C with the same efficiency as with the wild-type large T gene, but their growth was arrested after a shift to 40 degrees C, with a progressive loss in cell viability. This result indicates a continuous requirement for a large T function in the maintenance of "immortality."


[Differences in anatomical area-specific wound healing in hidradenitis suppurativa/acne inversa after surgery].
Hidradenitis suppurativa/acne inversa is an inflammatory, debilitating disease for which wide local excision of the affected area with secondary wound healing is considered the treatment of first choice for the inactive scarring form or after adequate anti-inflammatory medical treatment. In this study, we aimed to assess the duration of complete secondary wound healing after surgical intervention for hidradenitis suppurativa/acne inversa. Twenty-three surgical procedures in 17 consecutive patients (eight female, nine male) were evaluated for duration of secondary wound healing at axillary or anogenital/inguinal sites. To investigate the contribution of hair follicle bulge progenitor cells in wound re-epithelialization, tissue samples of lesional and perilesional skin were analysed for expression of the stem cell marker, cytokeratin 15 (CK15), and CD200, a marker for human follicular stem cells that resides in the bulge area. Initial wound size did not differ significantly between surgical wounds in the axillary (mean: 30.0 cm2 ± 5.4) and anogenital/inguinal (mean: 35.3 cm2 ± 5.7) region. However, healing time to complete wound closure was almost twice as fast in the anogenital/inguinal (mean: 132 days ± 10.4) than axilla area (mean: 254 days ± 39.1; p < 0.01). The accelerated wound healing in the anogenital/inguinal region was accompanied by significantly enhanced CK15 and CD200 expression, compared to axillary wounds (p < 0.05). The anogenital/inguinal region showed significantly faster secondary wound healing after surgical intervention for hidradenitis suppurativa/acne inversa compared to axillary wounds. We suspect differences in pilosebaceous unit density and thus hair follicle progenitor cells (as mirrored by CK15 and CD200 expression) to be the main driver behind this finding.


Relationships among fitness, body composition, and physical activity.
This study was designed to examine the associations of physical activity and body composition with cardiorespiratory fitness in eighth grade girls. A random sample of 1440 eighth grade girls at 36 schools participated in this cross-sectional investigation, which represented an ethnically and geographically diverse group. Cardiorespiratory fitness was assessed using a modified physical work capacity test on a cycle ergometer that predicted workload at a heart rate of 170 beats.min. Physical activity was assessed over 6 d in each girl using an accelerometer and body composition was estimated from body mass index and triceps skinfolds using a previously validated equation. Pearson correlations and multiple regression analyses were used to determine the relationships among fitness, physical activity, and body composition. Significant linear relationships among cardiorespiratory fitness, body composition, and physical activity were found. The combination of fat and fat-free mass along with racial group and a race by fat-free-mass interaction accounted for 18% (R) of the variation in physical fitness. Adding moderate-to-vigorous physical activity to the regression model increased the R to 22%. Black girls had somewhat lower fitness levels (P < 0.05) especially at higher levels of fat and fat-free mass than other racial/ethnic groups. Physical activity, fat-free mass, and the interaction between fat-free mass and racial group are significantly associated with cardiorespiratory fitness in adolescent girls.


Histologic classification of the combined nevus. Analysis of the variable expression of melanocytic nevi.
The designation combined nevus gives recognition to mixed cytologic patterns. In the common variant, plump, pigmented spindle cells form fascicles among nests of ordinary nevus cells. In other variants, one or several cellular components that share cytologic features with either a blue nevus or a Spitz nevus are represented. Ninety-five cases, 49% of which were of the common type, were studied. Grossly, most of the lesions were darkly pigmented papules or nodules. The clinical diagnosis in three-fourths of the cases was nevus, blue nevus, or melanoma. Fifteen percent had concomitant histologic features of melanocytic dysplasia, and most of these lesions were of the common type. For the common variant, the cytologic features, pattern of apparent infiltration, and variable representation of the features of a premalignant melanocytic dysplasia often mislead a pathologist in interpreting and predicting biologic potential. In combined nevi, the phenotypic diversity and genetic lability of melanocytic nevus cells is manifested.


Has stroke mortality stopped declining in Spain?
To analyse the changes in stroke mortality trends in Spain by autonomous community and by sex during the period 1980-2016, using joinpoint regression models. Mortality data were obtained from the Spanish National Statistics Institute. Crude and standardised rates were calculated for each Spanish autonomous community, and for each sex. Joinpoint analysis was used to identify the best-fitting points showing a statistically significant change in the trend. Joinpoint analysis enabled us to differentiate between communities in which mortality rates showed a continuous decline throughout the study period in both sexes (Asturias, Cantabria, Castile and Leon, Ceuta, and Melilla) or in men only (Extremadura). In men, in all those communities in which changes in the trend were observed (all but Aragon, the Balearic Islands, and Murcia, where rates remained stable), we observed an initial period of decline (ranging from -3.4% in Catalonia and Extremadura, to -6.0% in Madrid) and a final period where the trends diverged: mortality rates continued to fall in Andalusia, Aragon, the Balearic Islands, and Madrid, but began to stabilise in Castile-La Mancha and Murcia and to increase in the Canary Islands. In women, in those communities where changes were observed (all but Aragon, Murcia, and the Basque Country, where rates remained stable), we observed an initial period of decline (ranging from -3.1% in Catalonia to -6.4% in Navarre) and a final period where divergent trends were observed: rates continued to decline in Andalusia, Aragon, Catalonia, Galicia, Madrid, and the Basque Country, but began to stabilise in Extremadura and Murcia and to increase in the Canary Islands. Current data show that stroke mortality rates have decreased (in women in Andalusia), stabilised (in both sexes in Murcia, in men in Castile-La Mancha, and in women in Extremadura), and have even reversed (in both sexes in the Canary Islands). Further study is needed to identify the causes of these trends.


Substance P given intrathecally at the spinal T9 level increases arterial pressure and heart rate in the rat.
Administration of 10 micrograms of substance P intrathecally at the spinal T9 level of the unanaesthetized and the anaesthetized rat provoked an increase in arterial pressure and an increase in heart rate. Both cardiovascular responses began within 1-2 min of administration, and the peak of each occurred at 4-10 min. In the anaesthetized rat, which gave rise to the bulk of the responses reported, peak arterial pressure was ca 20 mm Hg greater than pre-administration levels, and peak heart rate was greater by ca 50 beats per min. Similar administration of vehicle failed to alter either parameter. Arterial pressure and heart rate in substance P-treated rats were significantly different from those in vehicle-treated rats up to 15-20 min after administration. Pretreatment with the sympathetic ganglion blocker, hexamethonium (10 mg/kg, i.v.), prevented the responses to intrathecal administration of substance P. Pretreatment with [D-Pro2, D-Phe7, D-Trp9]-substance P, an analogue with antagonist properties in the central nervous system, blocked both responses to substance P but failed to alter similar responses provoked by intrathecal administration of angiotensin II. Pretreatment with vehicle had no effect on responses to substance P or to angiotensin II. The antagonist also had partial agonistic effects. Both arterial pressure and heart rate were transiently increased, but this effect was reversed within 6 min; in the case of heart rate, values returned to the pre-application level but arterial pressure fell to a ca 15 mm Hg below this level. These results demonstrate a pharmacologically specific excitatory effect of substance P on spinal mechanisms controlling sympathetic output to the vessels and the heart; this output can be either via the adrenal medullae or via nerve pathways to the vessels and the heart. Our results also support the possibility that dysfunction of substance P systems at the spinal level may underly some models of hypertension and may be involved in some cases of essential hypertension in man, as well as in autonomic dysfunction associated with some neurological disorders.


Topical or oral antibiotics for children with acute otitis media presenting with ear discharge: study protocol of a randomised controlled non-inferiority trial.
Around 15%-20% of children with acute otitis media present with ear discharge due to a spontaneous tear or perforation of the eardrum (AOMd). Current guidance recommends clinicians to consider oral antibiotics as first-line treatment in this condition. The opening in the eardrum however should allow topical antibiotics to enter the middle ear directly. Local administration of antibiotics does not expose children to systemic side effects and may put less selective resistance pressure on bacteria. Evidence on the effectiveness of this approach in children with AOMd is lacking. A primary care-based, open, individually randomised, controlled, non-inferiority trial. The trial aims to recruit 350 children aged 6 months to 12 years with AOMd and ear pain and/or fever. Participants will be randomised to 7 days of hydrocortisone-bacitracin-colistin eardrops five drops three times daily or amoxicillin oral suspension 50 mg/kg body weight per day, divided over three doses. Parents will keep a daily diary of AOM symptoms, adverse events and complications for 2 weeks. In addition, they will record AOM recurrences, healthcare utilisation and societal costs for 3 months. The primary outcome is the proportion of children without ear pain and fever at day 3. Secondary outcomes include ear pain and fever intensity/severity; days with ear discharge; eardrum perforation at 2 weeks; adverse events during first 2 weeks; costs; and cost effectiveness at 2 weeks and 3 months. The primary analyses will be intention-to-treat and per-protocol analyses will be conducted as well. The medical research ethics committee Utrecht, The Netherlands has given ethical approval (17-400/G-M). Parents/guardians of participants will provide written informed consent. Study results will be submitted for publication in peer-reviewed medical journals and presented at relevant (inter)national scientific meetings. The Netherlands National Trial Register; NTR6723. Date of registration: 27 November 2017.


Augmenter of liver regeneration enhances the success rate of fetal pancreas transplantation in rodents.
Treatment of fetal pancreas (FP) isografts with insulin-like growth factor-I greatly improves the rate of conversion to euglycemia in diabetic rats. Complete knowledge of other factors that may facilitate the engraftment and function of FP in vivo is still embryonic. Augmenter of liver regeneration (ALR) is a newly described polypeptide growth factor found in weanling rat livers. ALR has trophic effects on regenerating liver. We studied the effects of in situ administration of this agent on FP isografts in rats. Streptozotocin-diabetic Lewis rats (blood glucose > 300 mg/dl) received 16 FP isografts transplanted intramuscularly. ALR was delivered from day 1 through day 14, in doses of 40 or 400 ng/kg/d. Animals were followed for 3 months with serial weights and blood glucose monitoring. These animals were compared with those treated with vehicle alone. Of the group treated with ALR at 40 ng/kg/day for 14 days, 89% (eight of nine) were euglycemic (P=0.0003). Of the group treated with ALR at 400 ng/kg/day for 14 days, 88% (seven of eight) were euglycemic (P=0.0007). Of the group treated with vehicle alone, none of the six were euglycemic. Euglycemia is defined here as glucose < 200 mg/dl for 3 days. Pathology of the intramuscular transplant site showed patches of islet tissue embedded in fat. These patches demonstrated insulin immunoreactivity. Diabetes was reversed in a significantly greater proportion of FP + ALR-treated recipients than those animals treated with vehicle alone. Local delivery of growth factors may be used as an adjunct to FP transplantation to improve the rate of success. This in situ model may be useful to further evaluate other soluble factors.


Distribution patterns of metastases in recurrent laryngeal nerve lymph nodes in patients with squamous cell esophageal cancer.
Esophageal cancers frequently metastasize to recurrent laryngeal nerve lymph nodes (RLNNs). Knowledge of anatomic metastatic RLNN (mRLNN) distributions is needed for lymph node dissection during esophageal cancer surgery. Among 189 patients with esophageal cancer who underwent esophagectomy between 2008 and 2010, 51 (27.0%) had mRLNN. Sixty-four mRLNNs (right, 31; left, 33) were identified via preoperative computed tomography (CT); more than 2 unilateral metastases and/or nodes with unclear boundaries were excluded. Anatomical characteristics, including vertical distance from the sternal notch level, circumferential angle to the mid-sagittal tracheal plane, and short and long axes, were measured via CT. Respective mean right and left vertical distances from the sternal notch were +2.0 ± 13.1 mm and -14.5 ± 23.8 mm (p = 0.0006). Left mRLNNs existed in vertically wider and inferior areas along recurrent laryngeal nerve, compared to right mRLNNs. The respective mean right and left circumferential angles around the trachea (from the anterior mid-sagittal plane) were 137.2 ± 11.2° and 94.3 ± 31.6° (p < 0.0001). Left mRLNNs were distributed more widely around the trachea, especially anteriorly. The short axes were larger for right mRLNNs than for left mRLNNs (8.6 mm vs. 6.8 mm, p = 0.026). Compared with the right side, left mRLNNs were smaller and had a vertically longer and circumferentially wider distribution. Careful attention should be given to the left side during curative RLNN dissection.


Antigenic thymus-epidermis relationships. Reactivity of a panel of anti-thymic cell monoclonal antibodies on human keratinocytes and Langerhans cells.
We have investigated on human skin the reactivity of a panel of 42 anti-thymus monoclonal antibodies (MCA) supplied by the Third International Workshop and Conference on Human Leucocyte Differentiation Antigens, Oxford, 1986. MCA of the first cluster of differentiation (CD1) define a group of surface molecules expressed by cortical thymocytes. Some of them (OKT6, M241 and Na1/34) have been shown to react in normal human skin with the epidermal Langerhans cells (LC). Twenty-two CD1 MCA were investigated in the present study. On normal human skin, 13 MCA reacted with LC in situ. This result suggests and confirms the heterogeneity of CD1 MCA. Recently, some of them were shown to recognize biochemically different molecules and/or epitopes of thymocytes. In addition, 20 anti-thymic epithelium MCA were tested on human skin. The MCA which only reacted with the thymic epithelial cell network (except Hassall's corpuscles) decorated only the epidermal basal cell layer. The MCA which reacted with all the thymic epithelial cells (including Hassall's corpuscles) decorated all the epidermal cell layers. These results confirm the heterogeneity of the thymic epithelial microenvironment and underline the antigenic similarities between the thymic epithelial structures and the different epidermal cell layers. The existence of bone-marrow-derived CD1-positive cells (thymocytes or LC) in an epithelial cell network (the thymus and the epidermis) focus the speculation around the immunological role of the epidermal basal cell layer in the T cell education and the exact lineage of the epidermal LC.


A solution-processed molybdenum oxide treated silver nanowire network: a highly conductive transparent conducting electrode with superior mechanical and hole injection properties.
We demonstrate the fabrication of solution-processed MoOx-treated (s-MoOx) silver nanowire (AgNW) transparent conductive electrodes (TCEs) utilizing low-temperature (sub-100 °C) processes. The s-MoOx aggregates around the AgNW and forms gauze-like MoOx thin films between the mesh, which can effectively lower the sheet resistance by more than two orders of magnitude. Notably, these s-MoOx-treated AgNW TCEs exhibit a combination of several promising characteristics, such as a high and broad transmittance across a wavelength range of 400 to 1000 nm, transmission of up to 96.8%, a low sheet resistance of 29.8 ohm sq(-1), a low haze value of 0.90%, better mechanical properties against bending and adhesion tests, and preferable gap states for efficient hole injection in optoelectronic applications. By utilizing these s-MoOx-treated AgNW TCEs as the anode in ITO-free organic light emitting diodes, promising performance of 29.2 lm W(-1) and 10.3% external quantum efficiency are demonstrated. The versatile, multi-functional s-MoOx treatment presented here paves the way for the use of low-temperature, solution-processed MoOx as both a nanowire linker and a hole injection interfacial layer for future flexible optoelectronic devices.