Systemic immunization induces protective CD4+ and CD8+ T cell-mediated immune responses in murine Listeria monocytogenes meningoencephalitis.
The immune mechanisms underlying immunization-induced protection of mice from lethal central nervous system (CNS) listeriosis were evaluated by immunohistochemistry, flow cytometry of leukocytes isolated from the brain, reverse transcription-polymerase chain reaction analysis of intracerebral (i.c.) tumor-necrosis factor-alpha, interferon-gamma, interleukin (IL)-2, IL-1 beta, IL-10, granulocyte/macrophage colony-stimulating factor, and inducible nitric oxide synthase mRNA expression, and T cell depletion experiments. The data demonstrate that active immunization of mice prior to an i.c. infection with Listeria monocytogenes prevents the development of a fatal necrotizing encephalitis and accelerates the recruitment of an increased number of alpha beta T cell receptor (TcR)+ CD4+ and CD8+ T cells, gamma delta TcR+ T cells, B cells, granulocytes and macrophages to the brain compared to non-immunized animals. In addition, immunization induced a pronounced activation of i.c. macrophages and microglial cells as shown by an increased expression of MHC class II antigens. In parallel, transcript levels for all cytokine mRNA analyzed were higher in the brains of immunized mice. The protective effects of immunization were completely abolished by depletion of CD4+, CD8+, or both T cell subsets. All groups of T cell-depleted immunized mice developed a fatal necrotizing encephalitis with an increased i.c. bacterial load. In addition, cytokine mRNA synthesis was significantly impaired. The severity of disease was only slightly different between CD4+, CD8+ and CD4+/CD8+ T cell depleted mice, indicating that both subsets of T cells are required for an effective i.c. immune response to L. monocytogenes. This is in marked contrast to systemic listeriosis, and points to CNS-specific features of the immune response.


A comparison of orocaecal transit times assessed by the breath hydrogen test and the sulphasalazine/sulphapyridine method in healthy beagle dogs.
Orocaecal transit time (OCTT) was assessed in six healthy beagles by means of the breath hydrogen test (BH2T) and the sulphasalazine/sulphapyridine method (SLZ) after the administration of a test meal of canned food mixed with sulphasalazine. Orocaecal transit time was defined as the time taken from the oral administration of the test meal to the time when the first portion of the meal reached the colon. In five of the dogs the OCTTs assessed by the BH2T were shorter than those measured by the SLZ method by 30, 15, 45, 30 and 45 minutes. However, the median OCTT assessed by the BH2T (135 minutes, range 120 to 195 minutes) was not significantly different from that measured by the SLZ (180 minutes, range 150 to 210 minutes) and was highly correlated with it (r = 0.94, P = 0.016). The sixth dog maintained baseline hydrogen and plasma sulphapyridine readings throughout the monitoring period and the OCTT could not be measured.


Histopathologic examination for Helicobacter pylori as a possible etiopathogenic factor in laryngeal carcinoma.
We investigated the presence of Helicobacter pylori in laryngeal cancer specimens to reveal whether or not this is a risk factor in the development of squamous cell carcinoma of the larynx. Sixty-nine total laryngectomy specimens with the pathologic diagnosis of squamous cell carcinoma and 30 laryngeal tissue samples that had been taken for the investigation of nonneoplastic (polyp, nodule) diseases were studied. Specimens of both tumor and control groups were stained with hematoxylin-eosin and modified Giemsa stains, and then they were examined under a light microscope. In both groups, H. pylori could not be found in any of the cases. The histologic examination of our series did not reveal any clue related to the possible etiologic association between H. pylori and squamous cell carcinoma of the larynx.


RAD6B overexpression confers chemoresistance: RAD6 expression during cell cycle and its redistribution to chromatin during DNA damage-induced response.
The HR6A and HR6B genes, homologs of the yeast RAD6 gene, encode ubiquitin conjugating enzymes that are required for postreplication repair (PRR) of DNA and damage-induced mutagenesis. We show here that consistent with its role as a PRR protein, HR6 protein (referred as RAD6) expression is cell cycle regulated, with maximal levels expressed in late S/G2 phases of the cell cycle. Exposure of MCF10A cells to adriamycin (ADR) causes enhancement in the levels of RAD6B mRNA and protein. Inclusion of actinomycin D abolishes both basal and ADR-induced RAD6B transcription indicating that ADR-induced effects on RAD6B transcription result from an increase in transcriptional activity rather than from regulation of RAD6B mRNA stability. The increase in RAD6 protein expression observed in ADR-treated cells is dependent upon transcription and de novo protein synthesis, as addition of actinomycin D and cycloheximide eliminated the induction effects. Using in vivo crosslinking experiments, we demonstrate that only a small proportion of RAD6 is associated with chromatin in untreated MCF10A cells. However, treatment with ADR or cisplatin is accompanied by a significant increase and redistribution of RAD6 to DNA, and RAD6, RAD18, PCNA, phosphohistone H3, as well as p53 proteins are all found in the DNA fractions. These findings suggest that although RAD6 protein is present in the nucleus, its recruitment to the chromatin appears to be modulated by DNA damage. Whereas MCF10A cells engineered to overexpress ectopic RAD6B are significantly more resistant to ADR and cisplatin as compared to empty vector-transfected cells, MCF10A cells stably transfected with antisense RAD6B display hypersensitivity to these damage-inducing drugs. Analysis of PRR capacities in cisplatin-treated MCF10A cells stably transfected with empty vector, RAD6B or antisense RAD6B showed that whereas RAD6B-overexpressing and vector control MCF10A cells possessed the ability to convert newly synthesized DNA to higher molecular weight species, MCF10A cells depleted of RAD6B are PRR-compromised. Although no human diseases have been linked to mutations in the PRR pathway genes, these data suggest that RAD6 may play an essential role in DNA damage tolerance and recovery via modulation of PRR, and that imbalances in the levels of RAD6 could lead to changes in drug sensitivity and damage-induced mutagenesis.


Expression and purification of the mitochondrial serine protease LACTB as an N-terminal GST fusion protein in Escherichia coli.
LACTB is a mammalian mitochondrial protein sharing sequence similarity to the beta-lactamase/penicillin-binding protein family of serine proteases that are involved in bacterial cell wall metabolism. The physiological role of LACTB is unclear. In this study we have subcloned the cDNA of mouse LACTB (mLACTB) and produced recombinant mLACTB protein in Escherichia coli. When mLACTB was expressed as an N-terminal GST fusion protein (GST-mLACTB), full-length GST-mLACTB protein was recovered by glutathione-agarose affinity chromatography as determined by MALDI-TOF mass spectrometry and immunoblotting. Expression of mLACTB as a C-terminal GST fusion protein or with either an N- or C-terminal His6-tag resulted in proteolytic degradation of the protein and we were not able to detect full-length mLACTB. Analysis of GST-mLACTB by Fourier transform infrared spectrometry revealed the presence of alpha-helices, beta-sheets and turns, consistent with a well-defined secondary structure. These results show that mLACTB can be expressed as a GST fusion protein in E. coli and suggest that GST-mLACTB was properly folded.


Comparison between Amulet and Watchman left atrial appendage closure devices: A real-world, single center experience.
Data reporting a head-to-head comparison between Amulet and Watchman devices are scarce. The aim of this study was to compare the Watchman™ versus Amulet™ left atrial appendage closure (LAAC) devices in a consecutive, industry-independent registry. Patients who underwent LAAC using Watchman or Amulet devices from January 2014 to December 2019 at the University Heart Center Lübeck, Lübeck, Germany were included in the present analysis. Primary endpoints included periprocedural complications (in-hospital death, pericardial tamponade, device embolization, stroke, major bleeding and vascular access complications), and complications during long-term follow-up (ischemic stroke, hemorrhagic stroke, thromboembolism, device thrombus, bleeding and death). After matching the patients for age (±5 years), gender, CHA2DS2Vasc score (±1) and HASBLED score (±1), each of the Watchman and the Amulet groups included 113 patients. Patients in the Amulet group had significantly more periprocedural complications (2.7% vs 10.6%, p = 0.029; respectively) and more major bleeding complications (0% vs 5.3%, p = 0.029; respectively). During long-term follow-up, the rate of events was comparable between the Watchman and Amulet groups (18.3% versus 20.8%, p = 0.729; respectively). Amulet LAAC device was associated with increased periprocedural complications as compared to Watchman LAAC device. On long-term follow-up, both devices showed comparable efficacy and safety.


Designing a quality assurance process for quality control of nuclear magnetic resonance metabolomics studies of human blood.
High-resolution magic angle spinning (HRMAS) nuclear magnetic resonance (NMR)-based metabolomics has demonstrated its utility in studies of biofluids for various diseases. HRMAS NMR spectroscopy is uniquely well suited for analyzing human blood samples because of the small quantity of samples and minimal preparation required. To develop this methodology into standardized clinical protocols, establishment of the method's quality assurance (QA) and evaluations of its quality control (QC) are critical. This study aims to assess the QA/QC measured from human blood specimens in the form of serum and plasma through within-subject and between-subject comparisons, as well as stability and consistency comparisons over several freezing-thawing cycles of sample storage conditions, and most importantly, the agreement of pooled control samples against individual samples.


Hip fracture care during the COVID-19 pandemic: retrospective cohort and literature review.
The goal of this study was to investigate whether the COVID-19 pandemic has affected hip fracture care at a Level I Trauma hospital. The secondary goal was to summarize the published hip fracture reports during the pandemic. A retrospective cohort study. Level I Trauma Center. Eighty-six operatively treated hip fracture patients age ≥65 years, occurring from January 17 to July 2, 2020. N/A. We defined 3 phases of healthcare system response: pre-COVID-19 (period A), acute phase (period B), and subacute phase (period C). The primary outcome was 30-day mortality. Clinical outcomes including time to surgery (TTS) and length of stay (LOS) were extracted from the electronic medical record. Twenty-seven patients from Period A, 27 patients from Period B, and 32 patients from Period C were included. The 30-day mortality was not statistically different. The mean TTS was 20.0 +/- 14.3 hours and was the longest in Period C (22.1 +/- 9.8 hours), but the difference was not statistically significant. The mean LOS was 113.0 +/- 66.2 hours and was longest in Period B (120.9 +/- 100.6 hours). However, the difference was not statistically significant. The 30-day mortality, TTS, and LOS were not statistically different across multiple phases of pandemic at a level 1 trauma center. Our results suggest that we successfully adapted new protocol changes and continued to provide evidence-based care for hip fracture patients. Our results were comparable with that of other authors around the world.


Orexin receptors in the developing piglet hypothalamus, and effects of nicotine and intermittent hypercapnic hypoxia exposures.
Orexin and its receptors (OxR1 and OxR2) play a significant role in arousal and sleep regulation. Using developing piglets, we aimed to determine the effects of nicotine and Intermittent Hypercapnic Hypoxia (IHH), alone or in combination, on orexin receptor expression in the hypothalamus. Four piglet groups were studied: control (n=14), nicotine (n=14), IHH (n=10) and nic+IHH (n=14). Applying immunohistochemistry for OxR1 and OxR2 expression, eight nuclei/areas of the hypothalamus: dorsal medial nucleus (DMN), arcuate nucleus (ARC), perifornical area (PFA), paraventricular nucleus (PVN), lateral hypothalamic area (LHA), ventral medial nucleus (VMN), supraoptic nucleus, retrochiasmatic part (SONr) and tuberal mammillary nucleus (TMN), were studied. Compared to controls, OxR1 and OxR2 were increased due to exposures, however this was region dependent. Nicotine increased OxR1 in the DMN (P<0.001) and SONr (P=0.036), and OxR2 in the DMN (P<0.001), VMN (P=0.014) and the TMN (P=0.026). IHH increased OxR1 in the DMN, PVN, VMN and SONr (P<0.01 for all), and OxR2 in DMN (P<0.001), PFA (P=0.001), PVN (P=0.004), VMN (P=0.041) and the TMN (P<0.001). The nic+IHH exposure increased OxR1 expression in all nuclei (TMN excluded) however, the changes were not significantly different from IHH alone. For OxR2, the increased expression after nic+IHH was significant compared to IHH in the DMN, ARC and SONr. These results show that nicotine increases orexin receptor expression in a region dependent manner. IHH induced increases were specific to arousal and stress related regions and nic+IHH results suggest that for OxR1, nicotine has no additive effect whereas for OxR2 it does, and is region dependent.


Hippocampal substrate of sensory associations.
The role of the hippocampus in the association of two conditional stimuli was evaluated using the conditioned rabbit NM preparation and a sensory preconditioning paradigm. Multiple-unit activity was recorded from field CA1 during preconditioning. Paired presentations of CS1 and CS2 resulted in increased excitability which was highly correlated with later performance during CS2 test sessions. Unpaired presentations evoked a response to stimulus onset often followed by an inhibitory period. Bilateral kainic acid lesions of CA1 had no deleterious effect on traditional CS-US conditioning but abolished the association of CS1 and CS2. These data suggest that the hippocampus may be vital to the association of sensory events. Further, the SPC paradigm could provide an excellent model for evaluating simple cognitive deficits.


Studies on the diversity of the distinct phylogenetic lineage encompassing Glomus claroideum and Glomus etunicatum.
Morphological and molecular characters were analysed to investigate diversity within isolates of the Glomus claroideum/Glomus etunicatum species group in the genus Glomus. The inter- and intra-isolate sequence diversity of the large subunit (LSU) rRNA gene D2 region of eight isolates of G. claroideum and G. etunicatum was studied using PCR-single strand conformational polymorphism (SSCP)-sequencing. In addition, two isolates recently obtained from Southern China were included in the analysis to allow for a wider geographic screening. Single spore DNA isolation confirmed the magnitude of gene diversity found in multispore DNA extractions. An apparent overlap of spore morphological characters was found between G. claroideum and G. etunicatum in some isolates. Analysis of the sequence frequencies in all G. etunicatum and G. claroideum isolates (ten) showed that four LSU D2 sequences, representing 32.1% of the clones analysed for multispore extraction (564) were found to be common to both species, and those sequences were the most abundant in four of the ten isolates analysed. The frequency of these sequences ranged between 23.2% and 87.5% of the clones analysed in each isolate. The implications for the use of phenotypic characters to define species in arbuscular mycorrhizal fungi are discussed. The current position of G. claroideum/G.etunicatum in the taxonomy of the Glomeromycota is also discussed.


Use of isothermal titration calorimetry to study surfactant aggregation in colloidal systems.
Isothermal titration calorimetry (ITC) is a general technique that allows for precise and highly sensitive measurements. These measurements may provide a complete and accurate thermodynamic description of association processes in complex systems such as colloidal mixtures. This review will address uses of ITC for studies of surfactant aggregation to form micelles, with emphasis on the thermodynamic studies of homologous surfactant series. We will also review studies on surfactant association with polymers of different molecular characteristics and with colloidal particles. ITC studies on the association of different homologous series of surfactants provide quantitative information on independent contribution from their apolar hydrocarbon chains and polar headgroups to the different thermodynamic functions associated with micellization (Gibbs energy, enthalpy and entropy). Studies on surfactant association to polymers by ITC provide a comprehensive description of the association process, including examples in which particular features revealed by ITC were elucidated by using ancillary techniques such as light or X-ray scattering measurements. Examples of uses of ITC to follow surfactant association to biomolecules such as proteins or DNA, or nanoparticles are also highlighted. Finally, recent theoretical models that were proposed to analyze ITC data in terms of binding/association processes are discussed. This review stresses the importance of using direct calorimetric measurements to obtain and report accurate thermodynamic data, even in complex systems. These data, whenever possible, should be confirmed and associated with other ancillary techniques that allow elucidation of the nature of the transformations detected by calorimetric results, providing a complete description of the process under scrutiny.


Mucocutaneous lymph node syndrome (Kawasaki disease).
Mucocutaneous lymph node syndrome has rarely been reported in the dental literature despite the orofacial features characteristic of the disease. A case is reported in which the cardinal signs were present: erythema of the oral mucosa, cervical lymphadenopathy, conjunctivitis, pyrexia, and desquamation of the skin of the hands and feet. In addition, hydrops of the gallbladder and cardiac disorder were also found. The patient may first seek treatment by the dental practitioner, by whom the diagnosis may be made. Attention is drawn to the possibility of cardiac abnormalities that may influence dental treatment.


Prognostic value of the combined preoperative plasma fibrinogen and systemic inflammatory indexes in ESCC patients.
The prognostic indexes based on the combination of preoperative fibrinogen and systemic inflammatory indexes may have greater predictive value in esophageal squamous cell carcinoma (ESCC). It was found that the predictive ability of F-NLR was more valuable than other systemic inflammatory indexes. The preoperative F-NLR score was closely related to the TNM stage, and could be used as an important independent prognostic index for patients with ESCC. Then the nomogram model constructed by F-NLR and TNM stage had higher prognostic ability than that of AJCC stage for ESCC patients. Preoperative F-NLR is a new independent prognostic index and a potential marker for treatment response monitoring in patients with ESCC.


Attention Allocation During Exploration of Visual Arrays in ASD: Results from the ABC-CT Feasibility Study.
Visual exploration paradigms involving object arrays have been used to examine salience of social stimuli such as faces in ASD. Recent work suggests performance on these paradigms may associate with clinical features of ASD. We evaluate metrics from a visual exploration paradigm in 4-to-11-year-old children with ASD (n = 23; 18 males) and typical development (TD; n = 23; 13 males). Presented with arrays containing faces and nonsocial stimuli, children with ASD looked less at (p = 0.002) and showed fewer fixations to (p = 0.022) faces than TD children, and spent less time looking at each object on average (p = 0.004). Attention to the screen and faces correlated positively with social and cognitive skills in the ASD group (ps < .05). This work furthers our understanding of objective measures of visual exploration in ASD and its potential for quantifying features of ASD.


Impact of the intraoral location on the rate of biofilm growth.
The aims of the present study were: a) to assess the impact of the intraoral location on the rate of biofilm growth, and b) to establish an in vivo biofilm model to examine intraoral biofilm growth. Eight healthy volunteers wore acrylic splints with 15 glass slabs each in the upper and lower jaws to build up plaque. After 48 h, the specimens were removed and stained using the vital fluorescence technique. Biofilm thickness was evaluated by confocal laser scanning microscopy (CLSM). The mean plaque thickness amounted to 77.6 +/- 29.1 microm on the buccal sites of the upper jaw and 71.9 +/- 26.3 microm on the buccal sites of lower jaw. On the palatal site a biofilm of 52.1 +/- 26.2 microm thickness was grown, which was significantly less compared with the other locations evaluated (p < 0.001). The results demonstrate that the in situ biofilm thickness on the buccal sites was similar irrespective of the location in the oral cavity. The new splint system described may be a useful tool for further standardised experimental studies regarding influences on growth and structure of intraoral biofilms.


Indocyanine green guided near-infrared fluorescence (ICG-NIRF) enhances vascular anatomy in robotic assisted DIEP flap harvest.
ICG-NIRF imaging has gained clinical acceptance lately. This technology can be particularly advantageous in the case of robotic flap harvest. This paper presents a new approach to deep epigastric pedicle dissection using ICG-NIRF in the setting of robotic assisted DIEP flaps.


Inner ear implants for experimental electrical stimulation of auditory nerve arrays.
Electrode arrays chronically implanted in the inner ear are gaining increased use for experimental studies of the auditory nervous system, as well as for studies related to development of improved auditory prostheses. Commercially available electrode arrays are designed for human use and thus may be unsuitable for experimental studies, particularly in small animals. This paper describes a simple, inexpensive method for making custom electrode arrays in a variety of configurations, suitable for animals ranging from small rodents to non-human primates.


Representative volume elements of strain/stress fields measured by diffraction techniques.
Finite-element modelling has been used to simulate local strains and stresses within free-standing polycrystalline slabs of W, Cu and W-Cu, heated with free or constrained boundaries. The elastic strain values in crystallites that satisfied the diffraction condition were used to simulate the lattice strain data that would be obtained from diffraction analysis, from which the average stresses within diffracting domains were computed. Comparison of direct-space stresses in the model with the average stresses determined from diffraction analysis shows that the representative volume elements (RVEs) required to obtain equivalent stress/strain values depend on the deformation mode suffered by the material. Further, the direct-space and diffraction stress values agree only under strict sampling and strain/stress uniformity conditions. Consequently, in samples where measurements are conducted in volumes smaller than the RVE, or where the uniformity conditions are not satisfied, further experimental and numerical techniques might be needed for the accurate determination of applied or residual stress distributions.


Histopathologic features and molecular genetic landscape of HER2-amplified endometrial carcinomas.
HER2 is an established therapeutic biomarker in advanced or recurrent endometrial serous carcinoma. Current clinical guidelines recommend HER2 testing exclusively in this endometrial carcinoma (EC) subtype; however, the full spectrum of ECs harboring HER2 amplification remains ill-defined. The present study characterizes the clinicopathologic and molecular features of HER2-amplified ECs across all histologic subtypes. Retrospective analysis of our institutional cohort of 2,042 ECs subjected to targeted clinical massively parallel sequencing identified 77 (3.8%) cases with HER2 amplification, a group comprised of serous (n = 29), endometrioid (low-grade, n = 2, high-grade, n = 1) and clear cell (n = 4) carcinomas, carcinosarcomas (n = 18) and high-grade ECs with ambiguous features (HGEC, n = 23). A co-existing TP53 mutation was identified in 94% (72/77) of HER2-amplified ECs. Other recurrent genetic alterations included amplification of CCNE1 (22%) and ERBB3 (10%), FBXW7 mutations or deletions (13%), and mutations in PIK3CA (40%) and PPP2R1A (13%). The HER2 immunohistochemistry score was 2+ or 3+ for all evaluable cases (n = 61). Apart from carcinosarcomas, which often showed lower HER2 expression, particularly in the sarcomatous component, HER2 immunohistochemical staining pattern and intensity were similar across EC subtypes. Intratumor heterogeneity in HER2 expression was common and correlated with genetic heterogeneity as detected by fluorescence in-situ hybridization. These results demonstrate the frequent co-occurrence of HER2 amplification with TP53 mutation and high-grade histology, rather than being specific to serous carcinoma, per se. Overall, these findings suggest that HER2 targeted therapy may be more broadly applicable to all high-grade EC histotypes and consideration should be given to expanding therapeutic eligibility.


Effect of platelet activating factor antagonist treatment on gentamicin nephrotoxicity.
To assess whether PAF could be involved in the gentamicin-induced nephrotoxicity, we have studied the effect of PAF antagonist BN-52021 on renal function in rats after gentamicin (GENTA) treatment. Experiments were completed in 21 Wistar rats divided into three groups: group GENTA was injected with gentamicin 100 mg kg(-1) body wt/day s.c. for 6 days. Group GENTA + BN received gentamicin and BN-52021 i.p. 5 mg kg(-1) body wt/day. A third group served as control. Rats were placed in meta-bolic cages and plasma creatinine and creatinine clearance were measured daily. GENTA group showed a progressive increase in plasma creatinine, a drop in creatinine clearance and an increase in urinary excretion of N-acetyl-beta-D-glucosaminidase and alkaline phosphatase. GENTA + BN group showed a lesser change in plasma creatinine and a creatinine clearance, but no difference with GENTA group in urinary excretion of NAG and AP were observed. Histological examination revealed a massive cortical tubular necrosis in rats treated with gentamicin, whereas in BN-52021 injected animals tubular damage was markedly attenuated. The present results suggest a role for PAF in the gentamicininduced nephro-toxicity.


Frequency and association of mitochondrial genetic variants with neurological disorders.
Mitochondria are small cytosolic organelles and the main source of energy production for the cells, especially in the brain. This organelle has its own genome, the mitochondrial DNA (mtDNA), and genetic variants in this molecule can alter the normal energy metabolism in the brain, contributing to the development of a wide assortment of Neurological Disorders (ND), including neurodevelopmental syndromes, neurodegenerative diseases and neuropsychiatric disorders. These ND are comprised by a heterogeneous group of syndromes and diseases that encompass different cognitive phenotypes and behavioral disorders, such as autism, Asperger's syndrome, pervasive developmental disorder, attention deficit hyperactivity disorder, Huntington disease, Leigh Syndrome and bipolar disorder. In this work we carried out a Systematic Literature Review (SLR) to identify and describe the mitochondrial genetic variants associated with the occurrence of ND. Most of genetic variants found in mtDNA were associated with Single Nucleotide Polimorphisms (SNPs), ~79%, with ~15% corresponding to deletions, ~3% to Copy Number Variations (CNVs), ~2% to insertions and another 1% included mtDNA replication problems and genetic rearrangements. We also found that most of the variants were associated with coding regions of mitochondrial proteins but were also found in regulatory transcripts (tRNA and rRNA) and in the D-Loop replication region of the mtDNA. After analysis of mtDNA deletions and CNV, none of them occur in the D-Loop region. This SLR shows that all transcribed mtDNA molecules have mutations correlated with ND. Finally, we describe that all mtDNA variants found were associated with deterioration of cognitive (dementia) and intellectual functions, learning disabilities, developmental delays, and personality and behavior problems.


[ESC guidelines 2019 on diagnostics and management of acute pulmonary embolism].
This article on the new European Society of Cardiology (ESC) guidelines for diagnostics and management of acute pulmonary embolism (PE) focusses on new or changed recommendations compared to the previous version of the guidelines from 2014. The current risk-adjusted management algorithm for acute PE includes the clinical severity, aggravating comorbid conditions and right ventricular dysfunction. For low-risk patients early discharge and outpatient treatment are possible, whereas for high-risk patients reperfusion treatment and hemodynamic support have to be considered, depending on the hemodynamic situation and contraindications in the individual patient. Effective therapeutic anticoagulation for at least 3 months is recommended for all patients with PE. Potential indicators for extended anticoagulation are given in the guidelines (class I or class IIa recommendations). New oral anticoagulants (NOAC) are the first choice for anticoagulation in preference to vitamin K antagonists (VKA); however, they are not recommended in patients with severe renal dysfunction, during pregnancy or lactation and in patients with antiphospholipid antibody syndrome. Furthermore, a new algorithm for the follow-up after acute PE is proposed in the guidelines. In cases of symptomatic persistent pulmonary hypertension (PH) the transfer to a specialized center is recommended.


Chrysanthemum morifolium Extract Ameliorates Doxorubicin-Induced Cardiotoxicity by Decreasing Apoptosis.
It is well known that the anthracycline anticancer drug doxorubicin (DOX) induces cardiotoxicity. Recently, Chrysanthemum morifolium extract (CME), an extract of the purple chrysanthemum flower, has been reported to possess various physiological activities such as antioxidant and anti-inflammatory effects. However, its effect on DOX-induced cardiotoxicity is still unknown. An 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT)assay revealed that 1 mg/mL of CME reduced DOX-induced cytotoxicity in H9C2 cells but not in MDA-MB-231 cells. A TUNEL assay indicated that CME treatment improved DOX-induced apoptosis in H9C2 cells. Moreover, DOX-induced increases in the expression levels of p53, phosphorylated p53, and cleaved caspase-3,9 were significantly suppressed by CME treatment. Next, we investigated the effect of CME in vivo. The results showed that CME treatment substantially reversed the DOX-induced decrease in survival rate. Echocardiography indicated that CME treatment also reduced DOX-induced left ventricular systolic dysfunction, and a TUNEL assay showed that CME treatment also suppressed apoptosis in the mouse heart. These results reveal that CME treatment ameliorated DOX-induced cardiotoxicity by suppressing apoptosis. Further study is needed to clarify the effect of CME on DOX-induced heart failure in humans.


Treatment results and prognostic factors for advanced squamous cell carcinoma of the larynx treated with concurrent chemoradiotherapy.
To review our experience with concurrent chemoradiotherapy (CCRT) for patients with advanced resectable squamous cell carcinoma (SCC) of the larynx and to evaluate the factors affecting survival and larynx preservation. Retrospective study. The records of 102 patients with stage III or IV resectable SCC of the larynx treated with CCRT between February 1994 and March 2009 were reviewed. Of 102 patients, 59 were treated with high-dose regimens, including cisplatin, 5-fluorouracil (5-FU), methotrexate, and leucovorin or docetaxel, cisplatin, and 5-FU, and 43 were treated with low-dose regimens, including carboplatin and uracil-tegafur or S-1. Radiotherapy was delivered 5 days a week using a single daily fraction of 1.8–2.0 Gray (Gy), to a total dose of 66.0–70.2 Gy. Overall survival (OS), disease-specific survival (DSS), and DSS with larynx preservation were estimated using Kaplan–Meier methods. The log-rank test and Cox proportional hazards regression were used to identify significant prognostic factors for DSS and DSS with larynx preservation. The 5-year OS and DSS for all patients treated with CCRT were 63.9 and 70.7 %, respectively. The 5-year DSS with larynx preservation was 54.1 %. On multivariate analysis, N stage, synchronous multiple primary cancers, and the contents of chemotherapy were significant predictors of OS for patients undergoing CCRT; T stage, N stage, and the contents of chemotherapy were significant prognostic factors for larynx preservation. The treatment method including the indication for CCRT may be determined by the contents of the chemotherapy and the T and N stages of laryngeal SCC. It is important to diagnose multiple synchronous primary cancers before CCRT.


Microbial synthesis of chitinase in solid cultures and its potential as a biocontrol agent against phytopathogenic fungus Colletotrichum gloeosporioides.
Antifungal activity of chitinase can be effectively utilized in biologic pest control strategies. Because solid-state cultivation has been termed a cost-effective means for fungal growth and metabolite production, chitinase production by Trichoderma harzianum was studied using wheat bran-based solid medium containing 1% colloidal chitin. Chitinase synthesis was found to be growth associated because maximum enzyme (5.4 U/g of dry substrate) and biomass production occurred at 72 h. Substrate moisture had a critical impact on chitinase production; five grams of medium having an initial moisture content of 68.4% when incubated for 72 h increased the enzyme yield to 9.3 U/g of dry substrate. Optimization of colloidal chitin concentration showed that improvements in chitinase yield and maximum activity were attained with a 2% (w/w) concentration. Supplementation of additional nitrogen sources also influenced enzyme production, and the best yield was obtained with yeast extract. The effect of crude chitinase on hyphal morphology of the phytopathogenic fungus Colletotrichum gloeosporioides was swelling as well as lysis of hyphal wall, depending on the age of the mycelium. Studies of pH and thermal stability showed that crude culture filtrate was active over pH 4.0-6.0 and retained about 48.2% activity after 40 min of incubation at 40 degrees C.


Micronucleus induction in erythrocytes of the Hyla pulchella tadpoles (Amphibia: Hylidae) exposed to insecticide endosulfan.
Endosulfan is a synthetic chlorinated and environmental genotoxic pesticide used worldwide for crop production. We used the micronucleus test in erythrocytes of Hyla pulchella tadpoles in order to develop an experimental model for detecting genotoxic effects of the synthetic chlorinated cyclodiene endosulfan. The frequency of micronuclei was examined in blood smears obtained from tadpoles exposed in vivo to three different concentrations 2.5, 5, and 10 microg/l of the compound and fixed at two sampling times 48 and 96 h. As a positive control larvae were exposed to 40 mg/l of cyclophosphamide. Results obtained here demonstrated the genotoxic effects of the commercial formulation endosulfan in the experimental model assayed.


Bioprocess considerations for T-cell therapy: Investigating the impact of agitation, dissolved oxygen, and pH on T-cell expansion and differentiation.
Adoptive T-cell therapy (ACT) has emerged as a promising new way to treat systemic cancers such as acute lymphoblastic leukemia. However, the robustness and reproducibility of the manufacturing process remains a challenge. Here, a single-use 24-well microbioreactor (micro-Matrix) was assessed for its use as a high-throughput screening tool to investigate the effect and the interaction of different shaking speeds, dissolved oxygen (DO), and pH levels on the growth and differentiation of primary T cells in a perfusion-mimic process. The full factorial design allowed for the generation of predictive models, which were used to find optimal culture conditions. Agitation was shown to play a fundamental role in the proliferation of T cells. A shaking speed of 200 rpm drastically improved the final viable cell concentration (VCC), while the viability was maintained above 90% throughout the cultivation. VCCs reached a maximum of 9.22 × 106 cells/ml. The distribution of CD8+ central memory T cells (TCM ), was found to be largely unaffected by the shaking speed. A clear interaction between pH and DO (p < .001) was established for the cell growth and the optimal culture conditions were identified for a combination of 200 rpm, 25% DO, and pH of 7.4. The combination of microbioreactor technology and Design of Experiment methodology provides a powerful tool to rapidly gain an understanding of the design space of the T-cell manufacturing process.


Environmental and intrinsic modulations of venous differentiation.
Endothelial cells in veins differ in morphology, function and gene expression from those in arteries and lymphatics. Understanding how venous and arterial identities are induced during development is required to understand how arterio-venous malformations occur, and to improve the outcome of vein grafts in surgery by promoting arterialization of veins. To identify factors that promote venous endothelial cell fate in vivo, we isolated veins from quail embryos, at different developmental stages, that were grafted into the coelom of chick embryos. Endothelial cells migrated out from the grafted vein and their colonization of host veins and/or arteries was quantified. We show that venous fate is promoted by sympathetic vessel innervation at embryonic day 11. Removal of sympathetic innervation decreased vein colonization, while norepinephrine enhanced venous colonization. BMP treatment or inhibition of ERK enhanced venous fate, revealing environmental neurotransmitter and BMP signaling and intrinsic ERK inhibition as actors in venous fate acquisition. We also identify the BMP antagonist Noggin as a potent mediator of venous arterialization.


Building capacity for Public Health 3.0: introducing implementation science into an MPH curriculum.
Many public health programs fail because of an inability to implement tested interventions in diverse, complex settings. The field of implementation science is engaged in developing strategies for successful implementation, but current training is primarily researcher-focused. To tackle the challenges of the twenty-first century, public health leaders are promoting a new model titled Public Health 3.0 where public health practitioners become "chief health strategists" and develop interdisciplinary skills for multisector engagement to achieve impact. This requires broad training for public health practitioners in implementation science that includes the allied fields of systems and design thinking, quality improvement, and innovative evaluation methods. At UNC Chapel Hill's Gillings School of Global Public Health, we created an interdisciplinary set of courses in applied implementation science for Master of Public Health (MPH) students and public health practitioners. We describe our rationale, conceptual approach, pedagogy, courses, and initial results to assist other schools contemplating similar programs. Our conceptual approach recognized the vital relationship between implementation research and practice. We conducted a literature review of thought leaders in public health to identify skill areas related to implementation science that are priorities for the future workforce. We also reviewed currently available training programs in implementation science to understand their scope and objectives and to assess whether any of these would be a fit for these priorities. We used a design focused implementation framework to create four linked courses drawing from multiple fields such as engineering, management, and the social sciences and emphasizing application through case studies. We validated the course content by mapping them to implementation science competencies in the literature. To date, there is no other program that provides comprehensive interdisciplinary skills in applied implementation science for MPH students. As of April 2018, we have offered a total of eleven sections of the four courses, with a total enrollment of 142, of whom 127 have been master's-level students in the school of public health. Using Kirkpatrick's Model, we found positive student reaction, learning, and behavior. Many students have completed applied implementation science focused practicums, master's papers, and special studies. A systematically designed interdisciplinary curriculum in applied implementation science for MPH students has been found by students to be a useful set of skills. Students have demonstrated the capability to master this material and incorporate it into their practicums and master's papers.


P2Y12 inhibitor monotherapy after complex percutaneous coronary intervention: a systematic review and meta-analysis of randomized clinical trials.
It remains unknown whether the recent trend of short dual antiplatelet therapy (DAPT) followed by P2Y12 inhibitor monotherapy can simply be applied to patients undergoing complex percutaneous coronary intervention (PCI). We performed a systematic review and meta-analysis to evaluate P2Y12 inhibitor monotherapy vs. conventional DAPT in patients undergoing complex PCI and non-complex PCI (PROSPERO: CRD42022335723). Primary endpoint was the 1-year Net Adverse Clinical Event (NACE). Among 5,323 screened studies, six randomized trials fulfilled the eligibility criteria. A total of 10,588 complex PCI patients (5,269 vs. 5,319 patients) and 25,618 non-complex PCI patients (12,820 vs 12,798 patients) were randomly assigned to P2Y12 inhibitor monotherapy vs. conventional DAPT. In complex PCI patients, P2Y12 inhibitor monotherapy was associated with a lower risk of NACE than conventional DAPT [Odds ratio (OR) 0.76, 95% confidence interval (CI) 0.63-0.91, P = 0.003], whereas in non-complex PCI patients, P2Y12 inhibitor monotherapy was associated with a trend toward lowering the risk of NACE (OR 0.86, 95% CI 0.72-1.02, P = 0.09). This meta-analysis across randomized trials demonstrated that a strategy of short DAPT followed by P2Y12 inhibitor monotherapy reduces the risk of 1-year NACE in patients undergoing complex PCI.


Vaginitis in Turkish women: symptoms, epidemiologic - microbiologic association.
To determine the relationship between clinical-epidemiologic findings and microbiologic results in vaginitis. A prospective study of 212 women with vaginal symptoms was undertaken. Each patient underwent an evaluation that included standardized history, interview after pelvic examination and vaginal culture. Only 45% of patients received diagnosis. Only two symptoms were more frequently noted with diagnosis. Itching was more frequent in vulvovaginal candidiasis (VCC) whereas odor was more frequent in Bacterial vaginosis (BV). BV was associated with a history of spontaneous and induced abortion, increased number of pregnancy, vaginal douching practice and intrauterine contraceptive device usage. VVC was found to be associated with no contraceptive usage and previous vaginitis history. Presenting symptoms alone or with microbiologic studies has limited value, and that of the women with vaginal symptoms may lack a microbiologic diagnosis. Evidence-based approaches that produce highly accurate diagnostic and effective treatment regimens would be expected in the future.


PFOA and PFOS diffusion through LLDPE and LLDPE coextruded with EVOH at 22 °C, 35 °C, and 50 °C.
Diffusion of perfluorooctanoate (PFOA) and perfluorooctane sulfonate (PFOS) through 0.1 mm and 0.75 mm LLDPE and 0.1 mm and 0.75 mm LLDPE coextruded with ethyl vinyl alcohol (denoted as CoEx) at room temperature (23 °C), 35 °C, and 50 °C is examined. These tests had negligible source depletion throughout the monitoring period, indicating limited contaminant partitioning and diffusion through the LLDPE. At 483 days, 23 °C receptor PFOA and PFOS concentrations, cr, were <8 μg/L (cr/co < 3.2 × 10-4) for all tests, and at 399 days elevated temperature receptor concentrations were < 0.4 μg/L (cr/co < 1.6 × 10-5) at 35 °C and <0.5 μg/L (cr/co < 2.0 × 10-5) at 50 °C for both PFOA and PFOS. LLDPE partitioning coefficient, Sgf was 0.9-1.4 (PFOA) and 2.8-5.3 (PFOS) based on sorption tests at 23 °C. Based on the best estimates of permeation coefficient, PgCoEx, for CoEx was consistently lower than PgLLDPE. For PFOA, CoEx had PgCoEx < 0.26 × 10-16 m2/s at 23 °C, <11 × 10-16 m2/s (35 °C), and < 10 × 10-16 m2/s (50 °C) while LLDPE had PgLLDPE < 3.1 × 10-16 m2/s (23 °C), <13 × 10-16 m2/s (35 °C), and <19 × 10-16 m2/s (50 °C). For PFOS, CoEx and LLDPE had PgCoEx < 0.55 × 10-16 m2/s and PgLLDPE < 3.2 × 10-16 m2/s (23 °C), PgCoEx < 8.3 × 10-16 m2/s and PgLLDPE < 40 × 10-16 m2/s (35 °C), and PgCoEx < 8.2 × 10-16 m2/s and PgLLDPE < 52 × 10-16 m2/s (50 °C). These values are preliminary and may change (e.g., decrease) as more data comes available over time. The Pg values deduced for PFOA and PFOS are remarkably lower than those reported for other contaminants of concern, excepting BPA, which exhibits similar behaviour.


The Effects of Daily Prednisone and Tocilizumab on Hemoglobin A1c During the Treatment of Giant Cell Arteritis.
To study the longitudinal effects of both glucocorticoids and tocilizumab, an interleukin-6 receptor inhibitor, on hemoglobin A1c (HbA1c ) levels during glucocorticoid tapering. We analyzed patients with complete data from the Giant Cell Arteritis Clinical Research Study (GiACTA) to investigate the impact of both glycemic and nonglycemic factors on changes in HbA1c levels over the 52-week trial. Giant cell arteritis (GCA) patients were randomized to receive either tocilizumab or placebo in addition to glucocorticoids. We used a multivariable mixed-effects model to evaluate associations of HbA1c level with daily glucocorticoid dose, randomization to receive tocilizumab, and red blood cell count in patients with and those without diabetes mellitus at baseline, over 52 weeks. In 209 patients, the median HbA1c level decreased by 0.50% (P < 0.01) in the group that received both tocilizumab and glucocorticoids (tocilizumab/glucocorticoid) and by 0.10% (P < 0.01) in the glucocorticoid-only group. Randomization to tocilizumab/glucocorticoid was associated with lower HbA1c (β = -0.209% in those without diabetes, P < 0.01; β = -0.290% in those with diabetes, P = 0.23). These changes had a sizable impact on glucose tolerance classification: 42.5% of patients in the tocilizumab/glucocorticoid group improved from prediabetes status to normal, compared to only 12.5% of patients treated with glucocorticoids alone. Daily glucocorticoid dose was associated with HbA1c level in patients with baseline diabetes (β = 0.018%/mg, P < 0.01) and those without baseline diabetes (β = 0.005%/mg, P < 0.01). Tocilizumab treatment was associated with a substantial reduction in HbA1c level, independent of glucocorticoid exposure, which may be achieved through a combination of glycemic and nonglycemic effects.


Toll-like receptor 2 expression on c-kit+ cells tracks the emergence of embryonic definitive hematopoietic progenitors.
Hematopoiesis in mammalian embryos proceeds through three successive waves of hematopoietic progenitors. Since their emergence spatially and temporally overlap and phenotypic markers are often shared, the specifics regarding their origin, development, lineage restriction and mutual relationships have not been fully determined. The identification of wave-specific markers would aid to resolve these uncertainties. Here, we show that toll-like receptors (TLRs) are expressed during early mouse embryogenesis. We provide phenotypic and functional evidence that the expression of TLR2 on E7.5 c-kit+ cells marks the emergence of precursors of erythro-myeloid progenitors (EMPs) and provides resolution for separate tracking of EMPs from primitive progenitors. Using in vivo fate mapping, we show that at E8.5 the Tlr2 locus is already active in emerging EMPs and in progenitors of adult hematopoietic stem cells (HSC). Together, this data demonstrates that the activation of the Tlr2 locus tracks the earliest events in the process of EMP and HSC specification.


Single-session photodynamic therapy combined with intravitreal bevacizumab for neovascular age-related macular degeneration.
To evaluate the efficacy of combined single-session photodynamic therapy (PDT) and intravitreal bevacizumab (IVB) for treatment of neovascular age-related macular degeneration (AMD). In a prospective interventional case series, patients with subfoveal choroidal neovascularization (CNV) underwent PDT followed by 1.25 mg IVB injection. Best-corrected visual acuity (BCVA) and optical coherence tomography (OCT) measurements were repeated at 6-week intervals and fluorescein angiography was performed after 12 weeks and when considered necessary thereafter. Repeat injections of IVB were performed based on fluorescein angiographic evidence of CNV leakage. Fourteen eyes were included in this study. Mean follow-up was 52.4+/-15.2 weeks (range: 26-74 weeks). Initially, mean BCVA was 0.80+/-0.42 logMAR and mean central macular thickness (CMT) was 308+/88 microm. At week 12, BCVA improved to 0.62+/-0.47 logMAR (p=0.006) and CMT reduced to 186+/-53 microm (p=0.003). Corresponding results were 0.53+/-0.52 logMAR (p=0.02) and 193+/-78 microm (p=0.002) after 24 weeks. A second IVB injection was performed in 13 eyes with a mean interval of 16.3+/-5.9 weeks. Combination therapy with single-session PDT and IVB can improve vision and reduce CMT in neovascular AMD. Repeat IVB injections may maintain the visual gain from the initial combination therapy.


Comparing Behavioral and Psychological Symptoms of Dementia and Caregiver Distress Caused Between Older Adults With Dementia Living in the Community and in Nursing Homes.
To investigate differences in behavioral and psychological symptoms of dementia (BPSD) and caregiver distress caused between older adults with dementia living in the community and in nursing homes. A comparative cross-sectional study. Participants were recruited from outpatient clinics of a tertiary psychiatric hospital and dementia units of a nursing home in Guangzhou, China. Neuropsychiatric Inventory was used to assess symptoms and caregiver distress. Dementia severity was determined using the Clinical Dementia Rating. This study included 157 community and 112 nursing home residents with dementia. Clinically significant symptoms (item score ≥ 4) were found in 88.5% of the former and 75% of the latter. Caregivers of 79.6% of the former and 26.8% of the latter reported that at least one of these caused them moderate-to-severe distress (distress score ≥ 3). Among the community patients, anxiety was the most frequent "very severe" symptom, while sleep disorders and agitation caused the most frequent "very severe" caregiver distress. After controlling for dementia severity and medication use, family caregiving remained an independent risk predictor for clinically significant symptoms and moderate-to-severe caregiver distress. The prediction of caregiver distress based on symptom scores varied across caregiver types and individual symptoms (R2 0.36-0.82). Group differences in clinically significant symptoms and moderate-to-severe caregiver distress showed at the stage of moderate-to-severe dementia. Tailored management strategies to relieve family caregivers' BPSD-induced distress are needed, especially at the stage of moderate-to-severe dementia. An effective service system should be established for supporting family caregivers to cope with BPSD.


SHED promote angiogenesis in stem cell-mediated dental pulp regeneration.
Dental pulp, plays an indispensable role in maintaining homeostasis of the tooth. Pulp necrosis always causes tooth nutrition deficiency and abnormal root development, which leads to tooth discoloration, fracture or even loss. Our previous study showed implantation of autologous SHED could regenerate functional dental pulp. However, the detailed mechanism of the implanted SHED participating in dental pulp regeneration remains unknown. In this study, we implanted SHED in a porcine dental pulp regeneration model to evaluate the regenerative effect and identify whether SHED promoted angiogenesis in regenerated dental pulp. Firstly we verified that xenogenous SHED had the ability to regenerated pulp tissue of host in vivo. Then we found the vasculature in regenerated pulp originated from implanted SHED. In addition, stem cells were isolated from regenerated dental pulp, which exhibited good multi-differentiation properties and promoted angiogenesis in pulp regeneration process and these results demonstrated that SHED promoted angiogenesis in stem cell-mediated dental pulp regeneration.


11 beta-Hydroxysteroid dehydrogenase activity in mesenteric arteries of spontaneously hypertensive rats.
1. 11 beta-Hydroxysteroid dehydrogenase (11-HSD) activity in mesenteric arteries of spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats was determined and expressed as the percentage conversion of [3H]-corticosterone to [3H]-11-dehydrocorticosterone. 2. 11-HSD activity was significantly decreased in mesenteric arteries of both 4 and 9 week old SHR (8.4 +/- 0.8%, 5.0 +/- 1.5%, respectively) compared with WKY rats (12.4 +/- 0.6%, 15.8 +/- 0.7%, respectively; P < 0.05). 3. Total RNA from rat vascular smooth muscle cells (VSMC) and endothelial cells (EC) were prepared with selective precipitation in 3 mol/L LiCl/6 mol/L urea. The expression of 11-HSD mRNA was confirmed in the rat VSMC but its mRNA expression was not detected in EC, using northern blot analysis. 4. The results in this study indicate that 11-HSD in the vascular wall may play a role in the pathogenesis of hypertension in SHR.


Vinblastine-C4 alkyl maleoyl and amino acid maleoyl derivatives: I. Chemistry and physicochemical data.
Vinblastine-C4 acyl derivatives were synthesized by linking alkyl maleoyl or amino acid maleoyl compounds through an ester linkage at the C4 position of the vindoline moiety of vinblastine. The condensation of the above mentioned compounds was performed by the mixed anhydride method. Their synthesis, yields and physiochemical data are reported.


An examination of the effects of detergents on anaerobic digestion.
An anaerobic filter was used to examine the treatability of wastewater formulated to simulate that from the manufacture of detergents. The detergent element was a mixture (1:1:1 v/v) of concentrated washing up liquid, a non-biological hand washing detergent and a fabric softener and, thus contained a combination of cationic surfactants, anionic surfactants, non-ionic surfactants and amphoteric surfactants. A concentration of 2 ml/l caused a deterioration in the performance of the digester which was pronounced after 7 days. When the packing of the filter was modified to include two layers of granular activated carbon, 320 g in total, the reactor was capable of treating a feed containing 10 ml/l. The amount of detergent removed during this second trial was greater than the Langmuir monolayer capacity of the carbon, indicating that both adsorption and degradation were occurring. The results also showed that, at detergent concentrations greater than 1 ml/l, the theoretical COD removal was lower than the actual COD removal. This was compatible with there being an element of adsorption by the activated carbon.


Interactions of the metal tolerant heterotrophic microorganisms and iron oxidizing autotrophic bacteria from sulphidic mine environment during bioleaching experiments.
Iron and sulfur oxidizing chemolithoautotrophic acidophilic bacteria, such as Acidithiobacillus species, hold the dominant role in mine environments characterized by low pH values and high concentrations of reduced sulfur and iron compounds, such as ores, rocks and acid drainage waters from mines. On the other hand, heterotrophic microorganisms, especially their biofilms, from these specific niches are receiving increased attention, but their potential eco-physiological roles have not been fully understood. Biofilms are considered a threat to human health, but biofilms also have beneficial properties as they are deployed in waste recycling and bioremediation systems. We have analyzed interactions of the metal tolerant heterotrophic microorganisms in biofilms with iron oxidizing autotrophic bacteria both from the sulphidic mine environment (copper mine Bor, Serbia). High tolerance to Cu(2+), Cd(2+) and Cr(6+) and the presence of genetic determinants for the respective metal tolerance and biofilm-forming ability was shown for indigenous heterotrophic bacteria that included strains of Staphylococcus and Rhodococcus. Two well characterized bacteria- Pseudomonas aeruginosa PAO1 (known biofilm former) and Cupriavidus metallidurans CH34 (known metal resistant representative) were also included in the study. The interaction and survivability of autotrophic iron oxidizing Acidithiobacillus bacteria and biofilms of heterotrophic bacteria during co-cultivation was revealed. Finally, the effect of heterotrophic biofilms on bioleaching process with indigenous iron oxidizing Acidithiobacillus species was shown not to be inhibitory under in vitro conditions.


Mutation spectrum of and founder effects affecting the PTS gene in East Asian populations.
The enzyme 6-pyruvoyl-tetrahydropterin synthase (PTPS, gene symbol: PTS) is involved in the second step of the de novo biosynthesis of tetrahydrobiopterin (BH4), which is a vital cofactor of nitric oxide synthases and three types of aromatic amino acid hydroxylases; the latter are important enzymes in the production of neurotransmitters. We conducted a study of PTS mutations in East Asia, including Taiwan, Mainland China, Japan, South Korea, the Philippines, Thailand and Malaysia. A total of 43 mutations were identified, comprising 22 previously reported mutations and 21 new discovered mutations. Among these, the c.155A>G, c.259C>T, c. 272A>G, c.286G>A and c.84-291A>G mutations were the most common PTS mutations in East Asia, while the c.58T>C and c.243G>A mutations were, respectively, specific to Filipinos and Japanese originating from Okinawa. Further studies demonstrated that each of the mutations listed above was in linkage disequilibrium to a specific allele of polymorphic microsatellite marker, D11S1347. These results suggest the presence of founder effects that have affected these frequent mutations in East Asia populations. In this context, D11S1347 should become one of the most reliable polymorphic markers for use in prenatal diagnosis among PTPS deficient families, especially where mutations are yet to be identified.


Three-year follow-up of oral leukoplakia after neodymium:yttrium aluminum garnet (Nd:YAG) laser surgery.
Oral leukoplakia is a relatively common lesion with a significant proportion of cases changing into cancer. Since most leukoplakias are asymptomatic, the primary objective of treatment must aim at the prevention of such malignant transformation. The main objectives of the study are to observe (1) the efficacy, safety and acceptability of the neodymium:yttrium aluminum garnet (Nd:YAG) laser in the management of oral leukoplakia; (2) the nature of postoperative complications, if any, associated with laser ablation, and (3) the 3-year prognosis of oral leukoplakia treated with laser. Twenty-eight patients with histologically proven leukoplakia were treated with Nd:YAG laser. From this study, we observed that the patients treated with Nd:YAG laser had only mild to moderate pain, swelling and restricted mouth opening, which peaked between 72 h and 1 week. In a majority of the patients the healing was prolonged, to a maximum of 5 weeks, and there was no clinical evidence of scarring in 26 out of 28 cases. In this study we achieved a cure rate of 92.86% in a 6-month period. Further follow-up after 3 years yielded almost the same result, except that one patient was not available for follow-up. We concluded that Nd:YAG laser is an effective device in the management of oral leukoplakia, which is one of the major pre-cancerous lesions in our country.


Long-term results of helium ion irradiation of uveal melanoma.
Between 1978 and 1988, 307 patients with uveal melanoma were irradiated using helium ions at Lawrence Berkeley Laboratory. The length of follow-up ranged from 1-115 months (median 42 months). The 5-year actuarial treatment results were: local control rate, 96.8%, determinate survival rate, 81%, freedom from distant metastases, 76%, eye retention rate, 83%, and risk of developing neovascular glaucoma, 36%. Long-term vision outcome was analyzed in 81 patients with a minimum follow-up of 5 years. Forty-seven percent of patients retained vision of 20/200 or better. The median change in vision was a loss of four lines on the standard eye chart. Thirty-eight percent of patients had visual acuity either improve or remain within two lines of their pretreatment vision. A multivariate analysis identified tumor size as the only independently significant risk factor affecting survival, development of neovascular glaucoma, or the risk of enucleation; no risk factor correlated with local recurrence. Tumor size, tumor-fovea distance, and pretreatment visual acuity were independently significant risk factors influencing vision outcome. These results confirm that helium ion irradiation is an effective treatment for uveal melanoma which combines high rates of local control, survival, and eye retention with a substantial likelihood of long-term vision preservation.


Congenital abnormalities associated with extrahepatic portal hypertension.
Congenital abnormalities were present in 12 out of 30 (40%) children with extrahepatic portal hypertension of unknown cause, but in only 2 out of 17 (12%) children with extnahepatic portal hypertension secondary to umbilical vein catheterization or omphalitis. The most frequent abnormalities in this series and in published reports were atrial septal defect, malformation of the biliary tract, and anomalous inferior vena cava. These findings are consistent with the view that some cases with extrahepatic portal hypertension are congenital in origin.


Selective suppression of renal Na+/H+ exchanger isoform-3 by prolonged stimulation of rats with adrenocorticotropic hormone.
Prolonged stimulation with adrenocorticotropic hormone (ACTH) causes hypertension and increases Na+ intake and urine output in humans and animals. However, its biochemical basis remains to be established. Since renal Na+/H+ exchanger isoforms (NHE) and the sodium pump play an important role in this condition, their levels were examined in rats stimulated with ACTH. Male Wistar rats received daily sc injection of ACTH (30 microg/100 g of body wt) for 4 d. Half of the ACTH-stressed rats were kept for four additional days without injection of ACTH (poststimulation). In a third group, the animals were treated with dexamethasone (50 microg/100 g of body wt) daily for 4 d. A fourth group consisted of unstressed control animals. Levels of NHE proteins were measured by Western blot analysis. Sodium pump activity was assessed by the level of ouabain-sensitive K-stimulated p-nitrophenylphosphatase activity (PNP) in the renal cortex. ACTH caused a selective decrease in NHE-3, but not of NHE-1 or alpha-actin levels. Interestingly, this ACTH-induced change was not duplicated in the animals treated with dexamethasone. Immunofluorescence data demonstrated that NHE-3 is located in the renal proximal tubules. PNP activity, on the contrary, was increased in both the ACTH-stimulated and dexamethasone-treated animals. More important, these changes in NHE-3 and PNP activity returned to the control level poststimulation. In conclusion, while PNP upregulation may be mediated by adrenocortical glucocorticoid, a role for glucocorticoids in the suppression of NHE-3 is less clear. These changes might impair renal tubular Na+ reabsorption and hence increase Na+ and water excretion in ACTH stimulation, thus acting as a counterbalance to normalize blood pressure in ACTH-stimulated animals.


Efficacy and Safety of Tadalafil 5 mg Administered Once Daily in Korean Men with Erectile Dysfunction: A Prospective, Multicenter Study.
The aim of this study was to evaluate the efficacy of a daily dose of tadalafil 5 mg as well as its safety for the cardiovascular system in men with erectile dysfunction. This study included a total of 162 men who were administered a daily dose of tadalafil 5 mg between April and December of 2009. A total of 127 men completed the 8-week clinical trial. The International Index of Erectile Function (IIEF)-5, blood pressure, and heart rate were measured before treatment with tadalafil (V1) and 4 (V2) and 8 weeks (V3) after treatment with tadalafil. Adverse effects were assessed at V1, V2, and V3. In cases in which the International Prostate Symptom Score (IPSS) was ≥8 at V1, maximal flow rate (Qmax) and postvoid residual volume (PVR) were measured. The IIEF-5 values were 11.25±3.18, 14.56±3.79, and 16.91±3.56 at V1, V2, and V3, respectively, with significant improvement (V1 vs. V2, p<0.001; V1 vs. V3, p<0.001). The IPSS values were 10.59±5.56, 9.07±6.06, and 8.15±6.10 at V1, V2, and V3, respectively, and the differences were statistically significant (V1 vs. V2, p<0.001; V1 vs. V3, p<0.001). There were no significant differences in blood pressure or heart rate. Adverse effects were observed in 7 men (5.51%) at V2 and in 5 men (3.94%) at V3. Tadalafil 5 mg administered once-a-day may be effective in improving erectile function. Adverse effects on the cardiovascular system may be minimal. In addition, it is believed that this may also be effective in improving voiding symptoms.


Expression of the neuronal calcium sensor protein family in the rat brain.
The neuronal calcium sensor proteins are members of the calcium-binding protein superfamily. They control localized calcium signalling on membranes and may make G-protein cascades sensitive to cytosolic calcium. The family members are recoverin (visinin, S-modulin), neuronal calcium sensor-1 (frequenin), hippocalcin, neuronal visinin-like protein-1 (visinin-like protein, neurocalcin-alpha), neuronal visinin-like protein-2 and neuronal visinin-like protein-3. Recoverin is expressed only in the retina and pineal gland. Using in situ hybridization, we mapped the expression of the other neuronal calcium sensor protein genes in the adult rat brain. Neuronal visinin-like protein-1 messenger RNA has a widespread distribution and is abundant in all brain areas except the caudate-putamen. Neuronal calcium sensor-1 gene expression is pan-neuronal. Neuronal calcium sensor-1 messenger RNA is present in the dendrites of hippocampal pyramidal and granule cells, suggesting a specific role in dendritic function. Hippocalcin and neuronal visinin-like protein-2 are mainly expressed in the forebrain and have similar expression patterns (neocortex, hippocampus and caudate-putamen). Neuronal visinin-like protein-3 has the most restricted expression; its highest expression level is in the cerebellum (Purkinje and granule cells). However, the neuronal visinin-like protein-3 gene is also expressed in many ventral nuclei throughout the fore- and midbrain, in the medial habenulae, and in the superior and inferior colliculi. The neuronal calcium sensor proteins are a relatively unexplored family of Ca(2+)-binding proteins. They are likely to be involved in many diverse areas of neuronal signalling. In this paper, we describe their expression in the rat brain as determined by in situ hybridization. As all five neuronal calcium sensor protein genes have distinctive expression patterns, they probably perform specific functions.


Posterior fossa meningiomas: surgical experience in 52 cases.
Early reports of the surgical management of posterior cranial fossa meningiomas (PCFM) yielded poor results with high rates of mortality and morbidity. With the advent of modern neuroimaging and microsurgical techniques the results of surgery have improved markedly, but despite these advances removal of these lesions remains a challenge. The results of the surgical treatment of PCF meningiomas were examined with the aim to identify particular features associated with increased mortality and morbidity. Of 713 patients with meningioma, 52 patients were identified with PCFM. Total macroscopic excision was achieved in 44 patients (84%). Postoperative complications occurred in 28 patients (54%) with permanent sequelae in 18 (35%). There were no mortalities in the immediate postoperative period. Follow-up ranged from 14 to 174 months (mean 42), tumour has recurred in 11 patients (21%) with a long-term mortality of 11%. At their latest follow-up 41 (79%) of patients achieved Glasgow outcome scores of 4 or 5. Total excision of tumour should remain the goal of treatment in patients with PCFM. Despite the recent advances in preoperative planning and surgical techniques, the morbidity associated with surgery remains significant. Notwithstanding, the majority of patients achieve a good outcome with surgical treatment.


Increase in permeability of Escherichia coli outer membrane by local anesthetics and penetration of antibiotics.
The MICs of several antibiotics (both hydrophobic and hydrophilic) which penetrate very poorly into intact Escherichia coli cells were found to be 2- to 10-fold decreased in the presence of low doses of various local anesthetics (procaine, dibucaine, tetracaine, chlorpromazine, and quinine). The concentrations of anesthetics necessary for this effect have no adverse effect on cell growth and are markedly lower than those concentrations used clinically.


Transgene-host cell interactions mediate significant influences on the production, stability, and function of recombinant canine FVIII.
Recombinant FVIII manufacturing is characterized by poor product stability and low yields. Codon-optimization of transgenes accelerates translation by exploiting the synonymous codon usage bias of a species. However, this can alter the performance of the final product. Additionally, the effects of transgene design across diverse cell types are not well understood and are of interest for next-generation protein and gene therapies. To investigate the effects of transgene design across different host cells, B-domain-deleted (BDD) and modified codon-optimized (CO-N6) transgenes were inserted via lentiviral delivery into cBOECs, HEK293T, and MDCK cells. The CO-N6 cFVIII transgene produced threefold more protein per transgene in HEK293T cells, and sixfold more protein in the two canine cell lines. However, pharmacokinetic analysis in hemophilia A dogs demonstrated that cFVIII produced from cBOECs transduced with the CO-N6 transgene had significantly reduced in vivo recovery. Furthermore, this product showed reduced in vitro stability and activity on thrombin activation versus the BDD product. This trend was reversed in HEK293T lines. Overall, our results demonstrate the need for an integrated approach that not only assesses protein expression levels but also considers the influence that host-cells have on preserving the molecular and biochemical properties of the naturally occurring FVIII.


Impaired insulin biosynthetic capacity in a rat model for non-insulin-dependent diabetes. Studies with dexamethasone.
These studies of a rat model for non-insulin-dependent diabetes mellitus (NIDDM) were performed to determine whether hyperglycemia occurs when capacity to synthesize insulin is exceeded. The neonatal streptozocin (STZ)-treated rat has acute hyperglycemia with marked destruction of pancreatic beta-cells, followed by gradual regeneration to 50-70% normal beta-cell number. At age 4 wk, fed serum glucose concentration is only mildly elevated relative to controls. With age, the rats become progressively hyperglycemic, and by 12 wk they have marked impairment of glucose-stimulated insulin release. In these studies, dexamethasone (0.125 mg/kg/day for 4 days) was administered to control and to STZ-treated animals to produce insulin resistance. The relationship between insulin biosynthesis and serum glucose concentrations was assessed. In control rats, response to dexamethasone was similar at both 4 and 12 wk. Serum glucose levels and pancreatic insulin concentration remained unchanged. Both insulin biosynthetic rates (as measured by 3H-leucine incorporation into proinsulin) and proinsulin mRNA levels increased twofold. STZ-treated rats at age 4 wk demonstrated mild hyperglycemia. Dexamethasone injection resulted in an increase in insulin biosynthesis and proinsulin mRNA in these animals, while serum glucose did not increase. STZ-treated rats at 12 wk showed more profound hyperglycemia (serum glucose 315 +/- 38 mg/dl versus control, 187 +/- 12 mg/dl). A marked rise in serum glucose (to 519 +/- 42 mg/dl) was observed after 4 days of dexamethasone injection. Pancreatic insulin content became severely depleted relative to saline-injected, STZ-treated animals, and there was no response of levels of proinsulin mRNA.


Polymorphisms of cancer stem cell marker gene CD133 are associated with susceptibility and prognosis of gastric cancer.
This study was aimed to investigate the associations between single nucleotide polymorphisms of cancer stem cell marker genes, CD44 and CD133, and susceptibility and prognosis of gastric cancer. Five single nucleotide polymorphisms in CD44 and CD133 genes were genotyped in 898 gastric cancer cases and 992 controls. The A/C or C/C genotypes of CD133 rs2240688 were associated with decreased risk of gastric cancer comparing with the A/A genotype (odds ratio: 0.81; 95% CI: 0.67-0.97; p = 0.023). The T allele of CD133 rs3130 predicted a worse survival for gastric cancer patients receiving tumorectomy (hazard ratio: 1.28; 95% CI: 1.04-1.58; p = 0.020), independent from tumor node metastasis stage, vessel invasion and postoperational chemotherapy. CD133 polymorphisms are promising biomarkers for genetic susceptibility and prognosis prediction of gastric cancer.


Long non-coding RNAs and their biological roles in plants.
With the development of genomics and bioinformatics, especially the extensive applications of high-throughput sequencing technology, more transcriptional units with little or no protein-coding potential have been discovered. Such RNA molecules are called non-protein-coding RNAs (npcRNAs or ncRNAs). Among them, long npcRNAs or ncRNAs (lnpcRNAs or lncRNAs) represent diverse classes of transcripts longer than 200 nucleotides. In recent years, the lncRNAs have been considered as important regulators in many essential biological processes. In plants, although a large number of lncRNA transcripts have been predicted and identified in few species, our current knowledge of their biological functions is still limited. Here, we have summarized recent studies on their identification, characteristics, classification, bioinformatics, resources, and current exploration of their biological functions in plants.


Glutamic acid residues of bacteriorhodopsin at the extracellular surface as determinants for conformation and dynamics as revealed by site-directed solid-state 13C NMR.
We recorded (13)C NMR spectra of [3-(13)C]Ala- and [1-(13)C]Val-labeled bacteriorhodopsin (bR) and a variety of its mutants, E9Q, E74Q, E194Q/E204Q (2Glu), E9Q/E194Q/E204Q (3Glu), and E9Q/E74Q/E194Q/E204Q (4Glu), to clarify contributions of the extracellular (EC) Glu residues to the conformation and dynamics of bR. Replacement of Glu-9 or Glu-74 and Glu-194/204 at the EC surface by glutamine(s) induced significant conformational changes in the cytoplasmic (CP) surface structure. These changes occurred in the C-terminal alpha-helix and loops, and also those of the EC surface, as viewed from (13)C NMR spectra of [3-(13)C]Ala- and [1-(13)C]Val-labeled proteins. Additional conformational changes in the transmembrane alpha-helices were induced as modified retinal-protein interactions for multiple mutants involving the E194Q/E204Q pair. Significant dynamic changes were induced for the triple or quadruple mutants, as shown by broadened (13)C NMR peaks of [1-(13)C]Val-labeled proteins. These changes were due to acquired global fluctuation motions of the order of 10(-4)-10(-5) s as a result of disorganized trimeric form. In such mutants (13)C NMR signals from Val residues of [1-(13)C]Val-labeled triple and quadruple mutants near the CP and EC surfaces (including 8.7-A depth from the surface) were substantially suppressed, as shown by comparative (13)C NMR studies with and without 40 micro M Mn(2+) ion. We conclude that these Glu residues at the EC surface play an important role in maintaining the native secondary structure of bR in the purple membrane.


From systemic lupus erythematosus to lupus nephritis: The evolving road to targeted therapies.
Systemic lupus erythematosus is a chronic autoimmune disease characterized by loss of tolerance against nuclear and cytoplasmic self-antigens, induction of immunity and tissue inflammation. Lupus nephritis (LN), the most important predictor of morbidity in SLE, develops in almost 30% of SLE patients at disease onset and in up to 50-60% within the first 10 years. Firstly, in this review, we put the pathogenic mechanisms of the disease into a conceptual frame, giving emphasis to the role of the innate immune system in this loss of self-tolerance and the induction of the adaptive immune response. In this aspect, many mechanisms have been described such as dysregulation and acceleration of cell-death pathways, an aberrant clearance and overload of immunogenic acid-nucleic-containing debris and IC, and the involvement of antigen-presenting cells and other innate immune cells in the induction of this adaptive immune response. This result in a clonal expansion of autoreactive lymphocytes with generation of effector T-cells, memory B-cells and plasma cells that produce autoantibodies that will cause kidney damage. Secondly, we review the immunological pathways of damage in the kidney parenchyma, initiated by autoantibody binding and immune complex deposition, and followed by complement-mediated microvascular injury, activation of kidney stromal cells and the recruitment of leukocytes. Finally, we summarize the rationale for the treatment of LN, from conventional to new targeted therapies, focusing on their systemic immunologic effects and the minimization of podocytary damage.


Pre-implantation embryos of Chinese hamster. II incidence and type of karyotype anomalies after treatment of the paternal post-meiotic germ cells with an alkylating mutagen.
Ninety-two male Chinese hamsters were treated with a single, sub-lethal dose of the alkylating cytostatic drug Trenimon. After 3--23 days they were mated with untreated females. The great majority of the male germ cells had been exposed to the mutagen while they were in the highly sensitive post-meiotic spermatid stage. The karyotypes of the resulting embryos were studied in the 4--8-cell stage. Out of 221 analysable embryos, 24.4% had aberrant karyotypes. Ploidy and genome mutations were, at 0.9% each, within control limits. Structural aberrations, involving one or several chromosomes, were present in 23.6% of the embryos (control 1.8%). 51% had a single aberrant centric element. The most frequent aberration types were deletions (54%), dicentrics (16%), translocations inversions and complex rearrangements with 22% and rings with 7%. About one-third of the cells, in addition, contained acentric fragments.


Stability in clinical use and stress testing of meropenem antibiotic by direct infusion ESI-Q-TOF: Quantitative method and identification of degradation products.
An ESI-MS/MS method through direct infusion was validated for quantitative analysis of meropenem powder for injection. The validation parameters were established in a rapid analysis of 30 s. Drug stability was studied through the submission to stress testing, resulting on four degradation products. Under hydrolytic conditions, in acid, neutral and alkaline media, the major degradation product was formed through the cleavage of the β-lactam ring. Oxidation of the drug using H2O2 (3%) showed the formation of two degradation products from a decarboxylation reaction and N-oxide formation. Under high humidity conditions, there was detected a dimer product. The stability of meropenem after reconstitution was studied in conditions that simulate its clinical use. In samples reconstituted and diluted in infusion fluids, an extensive degradation was observed. At room temperature meropenem maintained its content > 90% for up to 4 h when prepared in 5% glucose and for up to 12 h when prepared in 0.9% NaCl. Through ESI-MS/MS analyzes it was observed a degradation product formed by β-lactam ring cleavage, detected in all conditions studied. It was also identified a degradation product formed only in 5% glucose, generated by the hydrolysis of β-lactam followed by the attachment of a glucose molecule to the nitrogen of the pyrrolidine ring. In general, all the results obtained in the stability studies contribute to the knowledge about this antibiotic and future candidates of this class.


Purification of a 74-kilodalton surface glycoprotein from heart myoblasts that inhibits binding and entry of Trypanosoma cruzi into heart cells.
We have identified and purified a 74 kDa surface glycoprotein from heart myoblasts that specifically binds to Trypanosoma cruzi trypomastigotes, and inhibits the attachment and internalization of trypomastigotes into these cells. The native form of the 74 kDa glycoprotein was purified to apparent homogeneity by preparative scale isoelectrofocusing and anion exchange chromatography. Pre-incubation of trypomastigotes with soluble 74 kDa glycoprotein strongly inhibited the binding and internalization of trypomastigotes into heart myoblast monolayers in a concentration dependent-manner. Pre-incubation of heart myoblast monolayers with antibodies specific to the purified 74 kDa glycoprotein also strongly inhibited trypomastigote binding and internalization into heart cells in a concentration dependent manner. These results support the notion that the surface 74 kDa glycoprotein is a target molecule on heart myoblast cells to which T. cruzi binds in order to invade them.


A close contact identification algorithm using kernel density estimation for the ship passenger health.
COVID-19 has been spread globally, with ships posing a significant challenge for virus containment due to their close-quartered environments. The most effective method for preventing the spread of the virus currently involves tracking and physically isolating close contacts. In this paper, we propose the Close Contact Identification Algorithm (CCIA). The probability density of user location points may be higher in a certain spatial range such as a cabin where there are more location points. The characteristics of CCIA include using Kernel Density Estimation (KDE) to calculate the probability density of each user location point and seeking the maximum Euclidean distance between location points in each cluster for merging clusters. CCIA is capable of calculating the probability density of each location point, a feature that other clustering algorithms, such as Kmeans, Hierarchical, and DBSCAN, cannot achieve. The contribution of CCIA is using the probability density of each location point to identify close contacts in ship environments. The performance of CCIA shows more accurate clustering compared to Kmeans, Hierarchical, and DBSCAN. CCIA can effectively identify close contacts and enhance the capabilities of user devices in mitigating the spread of COVID-19 within ship environments.


Magnetic resonance imaging-based validation of the 2018 FIGO staging system in patients treated with definitive radiotherapy for locally advanced cervix cancer.
To validate the revised 2018 International Federation of Gynecologic and Obstetrics (FIGO) staging system in patients who underwent diagnostic magnetic resonance imaging (MRI) and radiotherapy (RT) for locally advanced cervix cancer. We analyzed 677 patients who were diagnosed with pelvic MRI and treated with definitive (chemo-)RT for locally advanced cervix cancer (stage IB2/IIA2-IVA or N+) between 1992 and 2018. Patients were classified according to 2009 and 2018 FIGO staging, and survival outcomes were compared. We developed a nomogram to improve prediction of progression-free survival (PFS). Pelvic and paraaortic lymph nodes were positive in 331 (48.9%) and 78 (11.5%) patients, respectively. At a median follow-up of 77.9 months, the 5-year PFS was 83.5%, 65.2%, 71.0%, 60.6%, 37.6% and 38.9% for IB, IIA, IIB, IIIA, IIIB and IVA according to FIGO 2009 and 88.9%, 60.0%, 73.8%, 66.7%, 36.3%, 68.9%, 43.6%, and 38.9% for IB, IIA, IIB, IIIA, IIIB, IIIC1, IIIC2, and IVA according to FIGO 2018, respectively. Survival of stage IIIC cervix cancer depended on the local extent of the tumor: the 5-year PFS of T1, T2, and T3 stages were 80.3%, 73.9%, and 45.5% for IIIC1 and 100%, 44.9%, and 23.4% for IIIC2. Histology, tumor size, node metastasis, FIGO 2009, and treatment modality were independent prognostic factors in the Cox regression analysis, and the nomogram incorporating these factors outperformed FIGO 2009 and FIGO 2018 (AUC 0.718 vs. 0.616 vs. 0.594). FIGO 2018 revision was associated with heterogenous outcomes among stage III cervix cancer patients. Our nomogram can assist the FIGO system in predicting PFS after definitive RT.


Are cellular polarisation and mitotic frequency prognostic factors for local recurrence in patients with ductal carcinoma in situ of the breast?
There is still no generally accepted histopathological classification system for ductal carcinoma in situ (DCIS) of the breast. Nuclear grade, with or without other histopathological parameters (i.e. comedo-type necrosis and cellular polarisation), has been demonstrated to yield prognostic information. A detailed method for the evaluation of the mitotic frequency in DCIS, based on an approach by Contesso, was used in this study. We also investigated if cellular polarisation and mitotic frequency were important for the ipsilateral local recurrence-free interval (IL-RFI) in 121 DCIS patients who had been operated upon with breast-conserving treatment (BCT) without radiotherapy. Both cellular polarisation and the mitotic frequency were associated with histopathological and cellular biological factors (in previous evaluations), and were of borderline significance for IL-RFI in the univariate analyses. However, when nuclear grade was included in the multivariate analyses (with or without the growth pattern), neither cellular polarisation nor the mitotic frequency were of any independent prognostic value.


Invasive cervical root resorption: treatment rationale with an interdisciplinary approach.
Invasive cervical resorption (ICR) is a significant and often aggressive pathologic process that, unfortunately, might lead to tooth loss. The presence of such a lesion in the cervical area is always a clinical challenge. This article presents an ICR case in which successful treatment was achieved by combining 4 disciplines: endodontics, orthodontics, periodontics, and prosthetics. Forced eruption combined with fiberotomy was used in this case for pulling the root rapidly from within the alveolar socket, thus exposing sound and healthy tooth material beyond the affected zone suitable for crown preparation. The presented interdisciplinary technique is offered for the prudent clinician as a solution in ICR cases of severe destructive nature.


Pregnancy Outcomes in the Tofacitinib Safety Databases for Rheumatoid Arthritis and Psoriasis.
Tofacitinib is an oral Janus kinase inhibitor for the treatment of rheumatoid arthritis (RA), and is being investigated for the treatment of psoriasis. Both conditions can present in women of child-bearing potential, but pregnancy was an exclusion and discontinuation criterion in tofacitinib randomized controlled trials (RCTs) because of the unknown effects of tofacitinib on mother and child. Tofacitinib is a small molecule that has the potential to cross the placenta. The objective was to report outcomes of pregnancy cases identified through April 2014 from tofacitinib RA/psoriasis RCTs, RA post-approval non-interventional studies, and spontaneous adverse-event reporting. Pregnancy outcomes were categorized as follows: healthy newborn, medical termination, fetal death, congenital malformation, spontaneous abortion, or pending/lost to follow-up. Out of 9815 patients, 1821 female patients of child-bearing age were enrolled in the RA/psoriasis RCTs; 47 women became pregnant, including 33 who received tofacitinib monotherapy, 13 who received combination therapy with methotrexate (RA patients only), and one patient whose therapy was still blinded. No fetal deaths were reported. One congenital pulmonary valve stenosis (monotherapy, n = 1), seven spontaneous abortions (monotherapy, n = 4; combination therapy, n = 3), and eight medical terminations (monotherapy, n = 4; combination therapy, n = 3; blinded therapy, n = 1) were identified. Remaining cases reported healthy newborns (n = 25) or were pending/lost to follow-up (n = 6). Forty-four cases of paternal exposure to tofacitinib were reported (monotherapy, n = 43; combination therapy, n = 1), including five spontaneous abortions (monotherapy, n = 4; combination therapy, n = 1), 23 healthy newborns, and 16 pending/lost to follow-up. The pregnancy outcomes reported in this small number of RA/psoriasis patients appear similar to those observed in the general population and in patients treated with biologic therapies for inflammatory diseases. However, definitive conclusions cannot be drawn, and pregnancy outcomes in patients receiving tofacitinib will continue to be monitored.


Effects of statin therapy in hospitalized adult COVID-19 patients: a systematic review and meta-analysis of randomized controlled trials.
COVID-19 is associated with endothelial activation and systemic inflammation; consequently, statins can be used in its treatment as they have anti-inflammatory, antithrombotic, and profibrinolytic properties and may interfere with COVID-19 viral entry into cells through disruption of cell membrane lipid rafts. We performed a meta-analysis of randomized clinical trials that compared statin therapy to placebo or to standard care in adult patients hospitalized for COVID-19. We searched the MEDLINE, EMBASE, and Cochrane Library databases for all-cause mortality, hospitalization duration, and admission to the intensive care unit. Of the 228 studies reviewed, four studies were included, with a total of 1,231 patients, of whom 610 (49.5%) were treated with statins. There was no significant difference in all-cause mortality (odds ratio [OR] 0.96; 95% confidence interval [95%CI]: 0.61-1.51; p=0.86; I2=13%), duration of hospitalization (mean difference [MD] 0.21; 95%CI: -1.74-2.16; p=0.83; I2=92%), intensive care unit admission (OR= 3.31; 95%CI: 0.13-87.1; p=0.47; I2=84%), need for mechanical ventilation (OR= 1.03; 95%CI: 0.36-2.94; p=0.95; I2=0%), or increase in liver enzyme levels (OR= 0.58; 95%CI: 0.27-1.25; p=0.16; I2=0%) between patients treated with or without statin therapy. Our findings suggest that in adult patients hospitalized with COVID-19, statin therapy results in no difference in clinical outcomes when compared to outcomes by placebo or standard of care. Prospero database registration: (www.crd.york.ac.uk/prospero) under the number CRD42022338283.


A Randomized Controlled Trial Comparing Microthin Descemet Stripping Automated Endothelial Keratoplasty With Descemet Membrane Endothelial Keratoplasty: Two-Year Report.
The purpose of this study was to report the 2-year outcomes of a double-blinded randomized controlled trial comparing Descemet membrane endothelial keratoplasty (DMEK) and microthin Descemet stripping automated endothelial keratoplasty (MT-DSAEK). Fifty-six eyes of 56 patients were randomized to DMEK or microthin DSAEK (MT-DSAEK). The main outcome measure was best spectacle-corrected visual acuity (BSCVA) at 24 months. Other secondary outcomes included complications, endothelial cell density, and vision-related quality-of-life (vQoL) scores. There was no statistically significant difference in BSCVA between the DMEK and MT-DSAEK groups at the 2-year time point (mean ± SD; 0.04 ± 0.14 vs. 0.12± 0.19, P = 0.061) in contrast to the 1-year results (mean ± SD; 0.04 ± 0.13 vs. 0.11 ± 0.09, P = 0.002) previously reported. Endothelial cell density did not show a statistically significant difference at 24 months between the DMEK and MT-DSAEK groups (1522 ± 293 cell/mm2 vs. 1432 ± 327 cells/mm2, P = 0.27). There were 2 additional graft rejection episodes in the MT-DSAEK group between the 1- and 2-year follow-up periods, but this did not result in graft failure. The mean vQoL scores between DMEK and MT-DSAEK indicated similar patient satisfaction between the groups (97.1 ± 4.0 vs. 92.6 ± 10.2, P = 0.13). In summary, the trial showed no significant difference in BSCVA at 24 months between the DMEK and MT-DSAEK groups. Both techniques continued to demonstrate comparable outcomes for complication rates, endothelial cell loss, and patient-reported vQoL scores. ISRCTN10578843.


Metabolic control and auditory information processing at altered glucose levels in insulin-dependent diabetes.
Previous work with a visual reaction time (RT) paradigm showed rate of mental processing to be slowed during a state of brain energy depletion (i.e., hypoglycemia). The present study employed an analog auditory RT paradigm to determine if modality differences in processing information might occur in response to glucose alteration. A balanced crossover design was used in which men with insulin-dependent diabetes completed RT tasks of increasing complexity at each of the following glucose levels: hypoglycemia (60 mg/dL), normoglycemia/control (110 mg/dL), and hyperglycemia (300 mg/dL). Results revealed two performance groups. A convergent group displayed slower RT responding during hypoglycemia, consistent with the visual RT pattern of results. A divergent group displayed better responding on one RT measure during hypoglycemia, and generally poorer responding on all RT measures during the other glucose conditions. Subjects in the divergent group maintained more stringent metabolic control and tended to have experienced more episodes of hypoglycemic unconsciousness. The present results provide the first evidence that more stringent metabolic control may be related to generally slower rates of processing auditory information. Possible explanations for these findings include hypotheses of immutable structual derangement, state-dependent performance effects, and a U-shaped response curve.


The Prevalence of Müllerian Anomalies in Women with a Diagnosed Renal Anomaly.
To characterize the prevalence of Müllerian anomalies (MAs) among patients with renal anomalies (RAs). DESIGN, SETTING, PARTICIPANTS, INTERVENTIONS, AND MAIN OUTCOME MEASURES: A retrospective chart review of female patients with RAs who presented to an academic pediatric hospital between 2007 and 2019 was performed. Patients were identified using International Classification of Diseases 9th and 10th revision codes. Data collected included the type of RA, presence and type of MA, method of diagnosis, and associated anomalies. RA subtype analysis was performed. We identified 5590 cases of RA for the years 2007 through 2019. A random, retrospective chart review was performed resulting in a study population of 363 RA patients. The prevalence of any MA in the overall RA population was 104/363 (29%) (95% confidence interval, 24% - 33%). The prevalence of MA for patients with renal agenesis was 59/182 (32%) compared with 45/181 (25%) for patients with renal dysgenesis. The most common MA were failures of Müllerian duct fusion. Only 73/352 (21%) of patients received screening for a MA at the time of RA diagnosis. Of patients without a diagnosed MA 187/259 (72%) were unscreened and either not yet menarchal or had unknown menarchal status. Of all RA patients, 29% (n = 104/363) had an underlying MA. No difference was found in the prevalence of MA in patients with renal agenesis vs dysgenesis. Limitations noted are that some patients might be of an age at which assessment of the Müllerian structures is suboptimal or who might not have been screened. These results suggest the need for a prospective study to determine evidence-based guidelines for screening for MA among patients diagnosed with any RA to avoid complications from an unrecognized MA.


Anaerobic metabolism and behavior during predatory encounters between snakes (Thamnophis elegans) and salamanders (Plethodon jordani).
1. We staged predatory encounters between snakes and salamanders in the laboratory. 2. Both snakes and salamanders accumulated significant quantities of lactic acid (0.3-2.3 mg/g) during predatory encounters. In a typical 14.4 min encounter, lactate concentration reached 260% of resting levels in snake predators and 880% of resting levels in salamander prey. This is one of the first demonstrations of anaerobiosis during natural 'burst' activity in both predator and prey. 3. Salamander responses to snake attack were surprisingly variable, and included writhing, thrashing, tail wrapping, tail autotomy, and biting. Antipredator responses were effective in nearly one third of trials. Lactate concentration was significantly correlated with duration of predatory encounters (r=0.57), snake mass, and salamander mass (Table 3). 4. Our calculations suggest that attack and ingestion of prey may cost snakes less than 1% of energy assimilated from prey.