Genotoxicity in lead treated human lymphocytes evaluated by micronucleus and comet assays.
Lead (Pb) which plays a significant role in modern industry is related to a broad range of physiological, biochemical, behavioural and genetical dysfunctions. Its exposure leads to an increased frequency of genetic aberrations in humans. Hence, this study was designed to assess the genotoxic effect of lead acetate at three dosage levels (10, 25 and 50 µg/mL) by employing: the Cytokinesis Block Micronucleus (CBMN) assay and the Comet assay in Peripheral Blood Lymphocyte Cultures. The results of this study revealed an increased level of DNA damage among treated groups. A significant increase in the tail length of comets and other indices was observed at 25 and 50 µg/mL concentrations comparatively. Thus, lead acetate induced single-strand breaks (SSB) and double strand breaks (DSB) in DNA, alkali-labile sites (ALS), oxidative DNA damage as well as DNA-DNA/DNA-protein/DNA-metal cross linking as evidenced by the Comet assay. The chromosome breakage, DNA misrepair, chromosome loss and telomere end fusion were determined by the Micronucleus assay. Micronucleus frequency in treated lymphocytes was significantly higher as compared to controls. Nucleoplasmic bridges increased significantly and Nuclear buds increased at higher two doses only in exposed cultures. Thus, these assays are better indices for lead induced genotoxicity and metal-nucleus interactions.


[Partial nephrectomy for renal cell carcinoma using a microwave tissue coagulator--postoperative recurrence and renal function].
Nowadays, partial nephrectomy is often selected for treatment of small renal cell carcinoma (RCC) because T1a RCC shows a favorable prognosis with a 5-year overall-survival rate exceeding 90%. To determine the methodological usefulness and treatment outcome of partial nephrectomy using microwave tissue coagulator (MTC), we evaluated our series of 121 patients (124 kidneys) with T1 RCC who underwent this procedure. Ninety-nine patients were elective and 22 patients were imperative cases. This procedure was indicated for asymptomatic solitary RCC; < or =4 cm in diameter in elective cases or < or =5 cm in imperative cases, expanding outward with a clear capsule, and showing neither renal pelvis nor renal sinus invasion. The median operation time was 150 minutes and the median blood loss was 217 ml. Eight patients needed transfusion, and conservative therapy cured 4 patients of postoperative urine leakage successfully. Three patients underwent early secondary nephrectomy due to multiple RCCs, invasive spindle cell carcinoma, or prolongation of urine leakage. No significant deterioration of renal function by creatinine clearance (Ccr) was observed, and postoperative Ccr could be precisely predicted from preoperative serum creatinine value and the renal parenchymal volume estimated with 3-dimensinal image reconstruction. Five-year overall-survival rate was 91% at the mean follow-up of up to 37.9 months and 107 patients were alive without disease. One patient showed local recurrence 2 years later and underwent radical nephrectomy. We believe that non-ischemic partial nephrectmy using MTC is a simple and reliable nephron-sparing surgery for small RCC in respect of postoperative recurrence and preservation of renal function.


[Clinical analysis of distal radius core decompression for chronic wrist pain].
To investigate the effectiveness of distal radius core decompression in the treatment of chronic wrist pain caused by various etiologies. A retrospective analysis was performed for the clinical data of 10 patients with chronic wrist pain treated with distal radial core decompression between January 2018 and December 2021. There were 6 males and 4 females with an average age of 37.4 years (range, 21-55 years). The disease duration ranged from 7 to 72 months, with an average of 26.5 months. Preoperative MRI examination showed that 10 cases had bone marrow edema at the distal radius on the affected side, and 8 cases had bone marrow edema in the carpal bones such as scaphoid and lunate bone. Among them, 3 patients had a history of wrist fracture, and 2 patients had Kienböck diseases (1 case each in stage ⅡB and stage ⅢA). Three cases were combined with triangular fibrocartilage complex (TFCC) type 1A injury. Two cases were combined with osteoarthritis, 1 of them was complicated with severe traumatic arthritis, the wrist arthroscopy showed that the TFCC was completely lost and could not be repaired, and the cartilage of the lunate bone and the ulnar head were severely worn.Visual analogue scale (VAS) score was used to evaluate the relief of wrist pain before operation, at 6 months after operation, and at last follow-up, and the range of motion of the affected wrist in dorsiflexion, palmar flexion, ulnar deviation, and radial deviation was measured. The degree of bone marrow edema was evaluated according to T1WI, T2WI, and STIR sequences of MRI. All the patients were followed up 12-22 months, with an average of 16.4 months. Except for 1 patient who experienced persistent wrist joint pain and limited mobility after operation, the remaining 9 patients showed significant improvement in pain symptoms and wrist joint mobility. The VAS score and range of motion of wrist dorsiflexion, palmar flexion, ulnar deviation, and radial deviation at 6 months after operation and at last follow-up were significantly improved when compared with those before operation, the VAS score and the range of motion of wrist ulnar deviation and radial deviation at last follow-up were further improved when compared with those at 6 months after operation, all showing significant differences ( P<0.05). There was no significant difference in wrist dorsiflexion and palmar flexion between at 6 months after operation and at last follow-up ( P>0.05). Bone marrow edema was improved in 6 patients on MRI at 6 months after operation, and was also improved in other patients at last follow-up. For chronic wrist pain caused by a variety of causes, distal radius core decompression can directly reduce the pressure of the medullary cavity of the distal radius, improve the blood supply of the corresponding distal structure, significantly alleviate chronic wrist pain, and provide an option for clinical treatment.


Genetic Modification of T Cells with Chimeric Antigen Receptors: A Laboratory Manual.
Redirected T cells genetically modified with a chimeric antigen receptor (CAR) have induced spectacular remissions of refractory leukemia/lymphoma in early phase trials, attracting interest to use CAR T cells in a variety of other applications including solid cancer and nonmalignant diseases. However, extensive preclinical explorations demand highly effective and robust procedures for the genetic modification of blood T cells; the same applies for engineering with a recombinant T cell receptor. We present laboratory procedures in a step-by-step protocol to engineer human and mouse T cells with a CAR by γ-retro- or lentiviral transduction for further preclinical testing.


Study protocol for a randomised double-blinded, sham-controlled, prospective, cross-over clinical trial of vagal neuromodulation for pain treatment in patients with chronic pancreatitis.
The management of chronic pancreatitis (CP) is challenging and requires a personalised approach focused on the individual patient's main symptoms. Abdominal pain is the most prominent symptom in CP, where central pain mechanisms, including sensitisation and impaired pain modulation, often are involved. Recent clinical studies suggest that vagal nerve stimulation (VNS) induces analgesic effects through the modulation of central pain pathways. This study aims to investigate the effect of 2 weeks transcutaneous VNS (t-VNS) on clinical pain in patients with CP, in comparison to the effect of sham treatment. Twenty-one patients with CP will be enrolled in this randomised, double-blinded, single-centre, sham-controlled, cross-over study. The study has two treatment periods: A 2-week active t-VNS using GammaCore device and a 2-week treatment with a sham device. During both treatment periods, the patients are instructed to self-administer VNS bilaterally to the cervical vagal area, three times per day. Treatment periods will be separated by 2 weeks. During the study period, patients will record their daily pain experience in a diary (primary clinical endpoint). In addition, all subjects will undergo testing which will include MRI, quantitative sensory testing, cardiac vagal tone assessment and collecting blood samples, before and after the two treatments to investigate mechanisms underlying VNS effects. The data will be analysed using the principle of intention to treat. The regional ethics committee has approved the study: N-20170023. Results of the trial will be submitted for publication in peer-reviewed journals. The study is registered at www.clinicaltrials.gov: NCT03357029.


A Multifunctional Pincer Ligand Supports Unsaturated Cobalt: Five Functionalities in One Pincer.
A pyridyl pincer ligand was developed to incorporate steric bulk, through a PtBu2 arm, and proton responsivity, through a pyrazole pincer ligand arm, together with reactivity at benzylic hydrogen and redox activity within a 1,4 diazabutadiene moiety. Binding it to CoCl2 yielded square-pyramidal [(PNNH)CoCl2 ], which was deprotonated by Li[N(SiMe3 )2 ] to form [{Li(THF)2 PNN}CoCl2 ]. Reduction of this LiCl adduct with KC8 under CO atmosphere led to formation of CoI mono- and dicarbonyl complexes, which can be protonated but also further deprotonated at the benzylic CH group to give a dearomatized pyridyl group. The ligand was characterized in its neutral, monoanionic, and dianionic forms, and the anions were shown to exist as intimate ion pairs with Li+ bound to pyrazolate N and chloride bound to Lewis acidic cobalt. X-ray photoelectron spectroscopy was used to assay both Li content and cobalt oxidation states. The general character of binding of LiCl to a metal complex acidic at metal and nucleophilic at ligand (pyrazolate Nβ) is discussed, as are potential catalytic applications of the concept.


Near-Infrared Fluorescent Probe with Large Stokes Shift for Imaging of Hydrogen Sulfide in Tumor-Bearing Mice.
Hydrogen sulfide (H2S) is an important endogenous gas signal molecule in living system, which participates in a variety of physiological processes. Very recent evidence has accumulated to show that endogenous H2S is closely associated with various cancers and can be regarded as a biomarker of cancer. Herein, we have constructed a new near-infrared fluorescent probe (DCP-H2S) based on isophorone-xanthene dye for sensing hydrogen sulfide (H2S). The probe shows remarkable NIR turn-on signal at 770 nm with a large Stokes shift of 200 nm, together with high sensitivity (15-fold) and rapid detection ability for H2S (4 min). The probe also possesses excellent selectivity for H2S over various other analytes including biothiols containing sulfhydryl (-SH). Moreover, DCP-H2S has been successfully applied to visualize endogenous and exogenous H2S in living cells (293T, Caco-2 and CT-26 cells). In particular, the excellent ability of DCP-H2S to distinguish normal mice and tumor mice is shown, and it is expected to be a powerful tool for detection of H2S in cancer diagnosis.


Polymorphisms in key pulmonary inflammatory pathways and the development of acute respiratory distress syndrome.
Acute Respiratory Distress Syndrome (ARDS) is an important clinical and public health problem. Why some at-risk individuals develop ARDS and others do not is unclear but may be related to differences in inflammatory and cell signaling systems. The Receptor for Advanced Glycation Endproducts (RAGE) and Granulocyte-Monocyte Stimulating Factor (GM-CSF) pathways have recently been implicated in pulmonary pathophysiology; whether genetic variation within these pathways contributes to ARDS risk or outcome is unknown. We studied 842 patients from three centers in Utah and 14 non-Utah ARDS Network centers. We studied patients at risk for ARDS and patients with ARDS to determine whether Single Nucleotide Polymorphisms (SNPs) in the RAGE and GM-CSF pathways were associated with development of ARDS. We studied 29 SNPs in 5 genes within the two pathways and controlled for age, sepsis as ARDS risk factor, and severity of illness, while targeting a false discovery rate of ≤ 5%. In a secondary analysis we evaluated associations with mortality. Of 842 patients, 690 had ARDS, and 152 were at-risk. Sepsis was the risk factor for ARDS in 250 (30%) patients. When controlling for age, APACHE III score, sepsis as risk factor, and multiple comparisons, no SNPs were significantly associated with ARDS. In a secondary analysis, only rs743564 in CSF2 approached significance with regard to mortality (OR 2.17, unadjusted p = 0.005, adjusted p = 0.15). Candidate SNPs within 5 genes in the RAGE and GM-CSF pathways were not significantly associated with development of ARDS in this multi-centric cohort.


Genetic and environmental correlational structure among metabolic syndrome endophenotypes.
Metabolic syndrome (MetS) is diagnosed by the presence of high scores on three or more metabolic traits, including systolic and diastolic blood pressure (SBP, DBP), glucose and insulin levels, cholesterol and triglyceride (TG) levels, and central obesity. A diagnosis of MetS is associated with increased risk of cardiovascular disease and type 2 diabetes. The components of MetS have long been demonstrated to have substantial genetic components, but their genetic overlap is less well understood. The present paper takes a multi-prong approach to examining the extent of this genetic overlap. This includes the quantitative genetic and additive Bayesian network modeling of the large TwinsUK project and examination of the results of genome-wide association study (GWAS) of UK Biobank data through use of LD score regression and examination of the number of genes and pathways identified in the GWASes which overlap across MetS traits. Results demonstrate a modest genetic overlap, and the genetic correlations obtained from TwinsUK and UK Biobank are nearly identical. However, these correlations imply more genetic dissimilarity than similarity. Furthermore, examination of the extent of overlap in significant GWAS hits, both at the gene and pathway level, again demonstrates only modest but significant genetic overlap. This lends support to the idea that in clinical treatment of MetS, treating each of the components individually may be an important way to address MetS.


The use of histology in 638 coronial post-mortem examinations of adults: an audit.
An audit was performed to determine the effectiveness of histological sampling of forensic post-mortem cases based on a review of three years' data, which comprised 638 adult autopsy cases. During the study period organs and tissues that appeared macroscopically normal and abnormal were extensively sampled. Histology was regarded as in some way contributory (providing, altering or confirming a cause of death) 53% of the time. The use of histology provided the cause of death in 49 (24%) of the 203 cases not given a cause of death after the completion of the macroscopic examination. When an interim cause of death had been supplied following the completion of the gross examination it was changed in 4.8% of cases, but there were no changes of the manner of death. The majority of the histological diagnoses or discrepancies involved the lungs and the heart. All diagnoses relevant to determining the cause of death would have been made if samples had been taken only from the left ventricle, right ventricle, coronary arteries, lungs, kidneys and brain with any tissue or organ that appeared abnormal macroscopically. A macroscopically identified abnormality that appeared to have been responsible for death was not sampled in 20 cases; consequently, more attention will be paid to sampling macroscopically abnormal tissues. As a result of this audit histology sampling practice has been revised and will be re-audited in the future.


[How I explore ... an orphan metastasis from melanoma].
A presumed melanoma metastasis is sometimes found without any evidence for the primary neoplasm. Such an orphan metastasis merits a complementary exploration. Searching for the primary melanoma must then be performed using non invasive and clinically oriented methods. Beside a thorough skin examination, otorhinolaryngological, ophtalmic and gynecologic searches are recommended. In addition, paraclinical investigations are conducted according to the metastatic location, the gender and the functional complaints of the patient. The prognosis of these orphan metastases is not more dramatic than when the primary lesion is identified. Therapies are similar to those proposed in the classical presentation of the disease.


Carotid sinus nerve transection abolishes the facilitation of breathing that occurs upon cessation of a hypercapnic gas challenge in male mice.
Arterial pCO2 elevations increase minute ventilation via activation of chemosensors within the carotid body (CB) and brainstem. Although the roles of CB chemoafferents in the hypercapnic (HC) ventilatory response have been investigated, there are no studies reporting the role of these chemoafferents in the ventilatory responses to a HC challenge or the responses that occur upon return to room air, in freely moving mice. This study found that an HC challenge (5% CO2, 21% O2, 74% N2 for 15 min) elicited an array of responses, including increases in frequency of breathing (accompanied by decreases in inspiratory and expiratory times), and increases in tidal volume, minute ventilation, peak inspiratory and expiratory flows, and inspiratory and expiratory drives in sham-operated (SHAM) adult male C57BL6 mice, and that return to room air elicited a brief excitatory phase followed by gradual recovery of all parameters toward baseline values over a 15-min period. The array of ventilatory responses to the HC challenge in mice with bilateral carotid sinus nerve transection (CSNX) performed 7 days previously occurred more slowly but reached similar maxima as SHAM mice. A major finding was responses upon return to room air were dramatically lower in CSNX mice than SHAM mice, and the parameters returned to baseline values within 1-2 min in CSNX mice, whereas it took much longer in SHAM mice. These findings are the first evidence that CB chemoafferents play a key role in initiating the ventilatory responses to HC challenge in C57BL6 mice and are essential for the expression of post-HC ventilatory responses.NEW & NOTEWORTHY This study presents the first evidence that carotid body chemoafferents play a key role in initiating the ventilatory responses, such as increases in frequency of breathing, tidal volume, and minute ventilation that occur in response to a hypercapnic gas challenge in freely moving C57BL6 mice. Our study also demonstrates for the first time that these chemoafferents are essential for the expression of the ventilatory responses that occur upon return to room air in these mice.


Secretory immunoglobulin A and cardiovascular reactions to mental arithmetic and cold pressor.
Secretory immunoglobulin A (sIgA) in saliva and cardiovascular reactions to mental arithmetic and cold pressor tasks were recorded in 16 healthy young men on two sessions, 4 weeks apart. Both tasks elicited significant increases in sIgA secretion rate, reflecting increases in both salivary volume and sIgA concentration. Whereas mental arithmetic elicited a mixed pattern of alpha- and beta-adrenergic cardiovascular reactions, the pattern of reactions to cold pressor was predominantly alpha-adrenergic. Task levels of sIgA secretion rate, sIgA concentration, and saliva volume showed moderate to high test-retest reliability (r = .52-.83), although test-retest correlations were less impressive for change scores (r = -.19-.53). The pattern of correlations between change in sIgA secretion rate and cardiovascular reactivity variables was inconsistent.


Patient-reported outcome measures in community-acquired pneumonia: a systematic review of application and content validity.
Patient-reported outcome measures (PROMs) are a vital component of patient-centred care. Community-acquired pneumonia (CAP) is a significant contributor to morbidity, mortality and health service costs globally, but there is a lack of consensus regarding PROMs for this condition. We searched MEDLINE, EMBASE and Cochrane Collaboration for studies, both interventional and observational, of adult recovery from CAP that applied at least one validated PROM instrument and were published before 31 December 2017. The full text of included studies was examined and data collected on study design, PROM instruments applied, constructs examined and the demographic characteristics of the populations measured. For all CAP-specific PROM instruments identified, content validity was assessed using the COnsensus based Standards for selection of health Measurement INstruments guidelines (COSMIN). Forty-two articles met the inclusion criteria and applied a total of 17 different PROM instruments including five (30%) classified as CAP specific, six (35%) as generic and six (35%) that measured functional performance or were specific to another disease. The 36-Item Short Form Survey (SF-36) was the most commonly used instrument (15 articles). Only one of 11 (9%) patient cohorts assessed using a CAP-specific instrument had a mean age ≥70 years. The CAP-Sym and CAP-BIQ questionnaires had sufficient content validity, though the quality of evidence for all CAP-specific instruments was rated as very low to low. PROM instruments used to measure recovery from CAP are inconsistent in constructs measured and have frequently been developed and validated in highly selective patient samples that are not fully representative of the hospitalised CAP population. The overall content validity of all available CAP-specific instruments is unclear, particularly in the context of elderly hospitalised populations. Based on current evidence, generic health instruments are likely to be of greater value for measuring recovery from CAP in this group.


Real-time Visualization and Analysis of Chondrocyte Injury Due to Mechanical Loading in Fully Intact Murine Cartilage Explants.
Homeostasis of articular cartilage depends on the viability of resident cells (chondrocytes). Unfortunately, mechanical trauma can induce widespread chondrocyte death, potentially leading to irreversible breakdown of the joint and the onset of osteoarthritis. Additionally, maintenance of chondrocyte viability is important in osteochondral graft procedures for optimal surgical outcomes. We present a method to assess the spatial extent of cell injury/death on the articular surface of intact murine synovial joints after application of controlled mechanical loads or impacts. This method can be used in comparative studies to investigate the effects of different mechanical loading regimens, different environmental conditions or genetic manipulations, as well as different stages of cartilage degeneration on short- and/or long-term vulnerability of in situ articular chondrocytes. The goal of the protocol introduced in the manuscript is to assess the spatial extent of cell injury/death on the articular surface of murine synovial joints. Importantly, this method enables testing on fully intact cartilage without compromising native boundary conditions. Moreover, it allows for real-time visualization of vitally stained articular chondrocytes and single image-based analysis of cell injury induced by application of controlled static and impact loading regimens. Our representative results demonstrate that in healthy cartilage explants, the spatial extent of cell injury depends sensitively on load magnitude and impact intensity. Our method can be easily adapted to investigate the effects of different mechanical loading regimens, different environmental conditions or different genetic manipulations on the mechanical vulnerability of in situ articular chondrocytes.


DNA packaging by L1 and L2 capsid proteins of bovine papillomavirus type 1.
Encapsidation of circular DNA by papillomavirus capsid protein was investigated in Cos-1 cells. Plasmids carrying both an SV40 origin of replication (ori) and an E. coli ori were introduced into Cos-1 cells by DNA transfection PV capsid proteins were supplied in trans by recombinant vaccinia viruses. Pseudovirions were purified from infected cells and their packaged DNA was extracted and used to transform E. coli as an indication of packaging efficacy. VLPs assembled from BPV-1 L1 alone packaged little plasmid DNA, whereas VLPs assembled from BPV-1 L1 + L2 packaged plasmid DNA at least 50 times more effectively. BPV-1 L1 + L2 VLPs packaged a plasmid containing BPV-1 sequence 8.2 +/- 3.1 times more effectively than a plasmid without BPV sequences. Using a series of plasmid constructs comprising a core BPV-1 sequence and spacer DNA it was demonstrated that BPV VLPs could accommodate a maximum of about 10.2 kb of plasmid DNA, and that longer closed circular DNA was truncated to produce less dense virions with shorter plasmid sequences. The present study suggests that packaging of genome within PV virions involves interaction of L2 protein with specific DNA sequences, and demonstrates that PV pseudovirions have the potential to be used as DNA delivery vectors for plasmids of up to 10.2 kb.


Surgical treatment of hilar strictures in hepatolithiasis.
The management of biliary strictures in hepatolithiasis, particularly at the hilus of the liver including the right and left hepatic bile ducts and their confluence, presents a challenge for surgeons because of their difficult access for surgical procedures and the life-threatening consequences, such as unrelenting cholangitis, recurrent biliary calculi, liver abscess and septicemia they can cause. This study analyzed the response of a group of patients with hilar strictures undergoing different surgical operations to assess methods dealing with this condition. A total of 545 patients with hepatolithiasis were admitted to our hospital. Among them, 256 patients were found to have an association of high bile duct strictures. These were classified into three types based on the sites of the stricture, and operated on with different combined surgical procedures in accordance with the type of the strictures. After the operation, further assessment was made during a follow-up period of between 12 months to 6 years. Postoperative morbidity and mortality were 15.7% (40/256) and 1.2% (3/256), respectively. Before discharge from the hospital, all patients underwent imaging investigation, and the retained stone rate was 20.3% (53/256). Of the 70% of patients (179/256) who were available for further assessment, 85% had good treatment results, were asymptomatic and required no medical treatment. A few patients got a poor result when symptoms of cholangitis occurred several times a year and required hospitalization. Successful treatment of hilar strictures in hepatolithiasis depends on the correct localization of the strictures, and complete exposure of the biliary tract proximal to strictures as well as the appropriate selection of the surgical procedures.


How to select a nanosimilar.
Nanomedicines in the class of nonbiological complex drugs (NBCDs) are becoming increasingly available. Up to 23 nanomedicines have been approved, and approximately 50 are in clinical development. Meanwhile, the first nanosimilars have entered the market through the generic approval pathway, but clinical differences have been observed. Many healthcare professionals may be unaware of this issue and must be informed of these clinically relevant variances. This article provides a tool for rational decision making for the inclusion of nanomedicines into the hospital formulary, including defined criteria for evaluation of substitutability or interchangeability. The tool was generated by conducting a roundtable with an international panel of experts and follows the same thought process that was developed and published earlier for the selection of biologicals/biosimilars. In addition to the existing criteria for biosimilars, a set of seven criteria was identified that specifically apply to nanosimilars. These include (1) particle size and size distribution, (2) particle surface characteristics, (3) fraction of uncaptured bioactive moiety, (4) stability on storage, (5) bioactive moiety uptake and (6) distribution, and (7) stability for ready-to-use preparation. Pharmacists should utilize their pharmaceutical expertise to use the appropriate criteria to evaluate the comparability of the drug to decide on interchangeability or substitutability.


Disinfection and removal of human pathogenic bacteria in arctic waste stabilization ponds.
Wastewater stabilization ponds (WSPs) are commonly used to treat municipal wastewater in Arctic Canada. The biological treatment in the WSPs is strongly influenced by climatic conditions. Currently, there is limited information about the removal of fecal and pathogenic bacteria during the short cool summer treatment season. With relevance to public health, the objectives of this paper were to determine if treatment in arctic WSPs resulted in the disinfection (i.e., removal of fecal indicator bacteria, Escherichia coli) and removal of selected human bacterial pathogens from the treated effluent. The treatment performance, with focus on microbial removal, was assessed for the one-cell WSP in Pond Inlet (Nunavut [NU]) and two-cell WSP in Clyde River (NU) over three consecutive (2012-2014) summer treatment seasons (late June-early September). The WSPs provided a primary disinfection treatment of the wastewater with a 2-3 Log removal of generic indicator E. coli. The bacterial pathogens Salmonella spp., pathogenic E. coli, and Listeria monocytogenes, but not Campylobacter spp. and Helicobacter pylori, were detected in the untreated and treated wastewater, indicating that human pathogens were not reliably removed. Seasonal and annual variations in temperature significantly (p < 0.05) affected the disinfection efficiency. Improved disinfection and pathogen removal was observed for the two-cell system in Clyde River as compared to the one-cell system in Pond Inlet. A quantitative microbial risk assessment should be performed to determine if the release of low levels of human pathogens into the arctic environment poses a human health risk.


The detection of myocardial viability and its clinical implications.
The mortality, morbidity and economic burden imposed by congestive heart failure are considerable. Despite a prevalence and prognosis comparable to the more commonly encountered cancers like breast and lung cancer, there has been relatively less emphasis in the western world on the screening, diagnosis and treatment of heart failure, at least until recently. This may be partly attributed to the fact that the complex pathophysiology of heart failure was ill understood and the heterogeneity of the syndrome not recognised, so that even among specialist cardiologists it was considered pretty much an irreversible condition amenable only to palliative therapy. However, our understanding of heart failure has grown significantly in the past decade. From the early days when all dysfunctional myocardium was considered necrosed, and revascularisation often dismissed on this basis, we have now progressed to the understanding that dysfunctional myocardium is a heterogeneous entity comprising necrotic, stunned, hibernating and myopathic myocardium, and that the therapeutic objectives in each of these categories is specific and should be addressed individually. In this article we briefly review the pathophysiology, and diagnostic and therapeutic implications of reversible ischaemic left ventricular dysfunction.


Monolayer-by-monolayer stacked pyramid-like MoS2 nanodots on monolayered MoS2 flakes with enhanced photoluminescence.
The precise control of the morphology and crystal shape of MoS2 nanostructures is of particular importance for their application in nanoelectronic and optoelectronic devices. Here, we describe a single step route for the synthesis of monolayer-by-monolayer stacked pyramid-like MoS2 nanodots on monolayered MoS2 flakes using a chemical vapor deposition method. First-principles calculations demonstrated that the bandgap of the pyramid-like MoS2 nanodot is a direct bandgap. Enhanced local photoluminescence emission was observed in the pyramid-like MoS2 nanodot, in comparison with monolayered MoS2 flakes. The findings presented here provide new opportunities to tailor the physical properties of MoS2via morphology-controlled synthesis.


Rhabdomyosarcoma of the urinary bladder with intraepithelial spread in an adult.
Rhabdomyosarcoma of the urinary bladder in adults is exceedingly rare. Reported cases have been of the embryonal botryoid type, as seen in children. We will describe a case of pleomorphic rhabdomyosarcoma occurring in the urinary bladder of an adult. In this case, we noted striking intraepidermal migration of tumor cells, resembling the epidermotropic growth characteristic of tumor cells in Paget disease of the breast. The pathological features of this apparently unique case form the subject of this report.


The unfolded protein response of B-lymphocytes: PERK-independent development of antibody-secreting cells.
When B-lymphocytes differentiate into plasma cells, immunoglobulin (Ig) heavy and light chain synthesis escalates and the entire secretory apparatus expands to support high-rate antibody secretion. These same events occur when murine B-cells are stimulated with lipopolysaccharide (LPS), providing an in vitro model in which to investigate the differentiation process. The unfolded protein response (UPR), a multi-pathway signaling response emanating from the endoplasmic reticulum (ER) membrane, allows cells to adapt to increasing demands on the protein folding capacity of the ER. As such, the UPR plays a pivotal role in the differentiation of antibody-secreting cells. Three specific stress sensors, IRE1, PERK/PEK and ATF6, are central to the recognition of ER stress and induction of the UPR. IRE1 triggers splicing of Xbp-1 mRNA, yielding a transcriptional activator of the UPR termed XBP-1(S), and activation of the IRE1/XBP-1 pathway has been reported to be required for expansion of the ER and antibody secretion. Here, we provide evidence that PERK is not activated in LPS-stimulated splenic B-cells, whereas XBP-1(S) and the UPR transcriptional activator ATF6 are both induced. We further demonstrate that Perk-/- B-cells develop and are fully competent for induction of Ig synthesis and antibody secretion when stimulated with LPS. These data provide clear evidence for differential activation and utilization of distinct UPR components as activated B-lymphocytes increase Ig synthesis and differentiate into specialized secretory cells.


Determination of membrane protein structure and dynamics by magic-angle-spinning solid-state NMR spectroscopy.
It is shown that molecular structure and dynamics of a uniformly labeled membrane protein can be studied under magic-angle-spinning conditions. For this purpose, dipolar recoupling experiments are combined with novel through-bond correlation schemes that probe mobile protein segments. These NMR schemes are demonstrated on a uniformly [13C,15N] variant of the 52-residue polypeptide phospholamban. When reconstituted in lipid bilayers, the NMR data are consistent with an alpha-helical trans-membrane segment and a cytoplasmic domain that exhibits a high degree of structural disorder.


Dimensions associated with successful implementation of a hospital based integrated order entry system.
Implementation of an integrated electronic medical record requires direct physician order entry. This application involves multi-level changes in the whole system of care, from physicians attitudes to interdepartmental relations. This study reports the results of the first round of a modified Delphi, where a diverse group of individuals were asked to identify the most important facilitating and impeding factors associated with implementation of an order entry application. From a Q-sort of their responses, we identified 20 systemic, behavioral, and attitudinal dimensions perceived to be causal factors in successful implementation. We also explored how these dimensions may influence success by comparing successful with unsuccessful hospitals in terms of the frequency with which these dimensions were differently mentioned by respondents. We found that although available functionality was the most commonly mentioned factor by all participants, hardware availability, physician involvement, administration support, and medical administration involvement were more often mentioned by successful hospitals than by less successful hospitals. These results suggest that these factors were not present in the less successful hospitals. We also found that the frequency of responses within each category varied depending on the institutional role of the individuals responding. Those involved in support tended to see organizational variables as more important than those in clinical positions, whereas clinicians viewed administrative support and involvement of the chief as more important. These findings support the notion that the changes involved in instituting a physician order entry system are system wide and involve individual as well as organizational factors.


Impact of the complement lectin pathway on cytomegalovirus disease early after kidney transplantation.
This study retrospectively investigated the association between pre-transplant levels of mannose-binding lectin (MBL) plus the associated serine protease (MASP)-2 and the occurrence of cytomegalovirus (CMV) infection and symptomatic CMV disease during the first 12 weeks after kidney transplantation. Materials and methods. Altogether 159 consecutive single kidney transplant recipients were included. The patients were screened for CMV pp65 antigenaemia every second week. No CMV prophylaxis or pre-emptive treatment was given. MBL and MASP-2 were measured in samples taken at transplantation and 10 weeks later. CMV infection, defined as at least one positive test, was found in 95 patients (59.8%). MBL and MASP-2 measured at transplantation were similar in patients with and without CMV infection. The incidence of CMV infection was also similar in 36 patients (58.3%) with pre-transplant MBL levels below the reference level (500 microg/L) and in patients with higher MBL levels (60.2%). Symptomatic CMV disease was diagnosed in 35 patients (22%), and MASP-2 levels at transplantation in the lower quartile range (<or=148 microg/L) was significantly associated with CMV disease during the first 12 weeks, P = 0.028. MBL levels decreased significantly from transplantation to 10 weeks later, and median (interquartile range) fell from 2597 (526-4939) microug/L to 1520 (270-3069) microg/L (P < 0.001). In contrast, MASP-2 levels increased significantly from 252 (148-382) microg/L to 380 (302-492) microg/L (P < 0.001). Pre-transplant MBL levels do not influence the incidence of any CMV infection or symptomatic CMV disease during the first 12 weeks after kidney transplantation. However, low MASP-2 levels may play a role in the development of symptomatic CMV disease.


Pedigree analysis of two brothers with severe oligozoospermia caused by maternal inv(X) (p22.3, q22) chromosome abnormality.
Sex chromosome abnormality (SCA) is one of the major causes of male spermatogenesis dysfunction. In our study, we sought to investigate the novel X chromosome inversion leading to severe oligozoospermia. Here, we report two brothers with severe oligozoospermia without any other abnormal clinical phenotype. The chromosome karyotypes in peripheral blood of both brothers were 46, Y, inv (X) (p22.3, q22), and no Y chromosome microdeletion was found. The karyotype of their mother was 46, X, inv (X) (p22.3, q22) and that of their father was 46, XY. This is the first report in China that X chromosomal inversion, 46, Y, inv (X) (p22.3, q22), is associated with severe oligozoospermia. This inversion may be a direct genetic risk factor for spermatogenesis.


A MICROSATELLITE ASSESSMENT OF SNEAKED FERTILIZATIONS AND EGG THIEVERY IN THE FIFTEENSPINE STICKLEBACK.
Attempts by males to steal fertilizations from other males are common in many species. In some sticklebacks, males also are known to steal eggs from the nests of rivals and to carry them back to their own nests. However the genetic consequences of these nest-raiding behaviors seldom have been investigated. Here we assess genetically the prevalence of sneaked fertilizations and egg stealing, and we describe the mating system in a natural population of the fifteenspine stickleback. Six microsatellite markers were developed and employed to assay a total of 1307 embryos from 28 nests. Guardian males and all nest-holding males in the local area also were genotyped for two to six loci Analysis of male genotypes and those of embryos revealed that five of the 28 nests (18%) contained progeny from sneaked fertilizations, and that four of the 24 nests (17%) with resident males contained stolen egg clutches Comparisons of the composite DNA genotypes of nest-holding males against those of inferred sneakers implicated one nest holder as the sneaker of a nest seven meters from his own. Also, the genetic data demonstrated that nests of males frequently contain eggs from multiple females. The multilocus genotypes of inferred mothers indicated that females mate with multiple males, sometimes over distances greater than one kilometer.


Serum osteocalcin levels in hyperthyroidism before and after antithyroid therapy.
Hyperthyroidism is characterized by accelerated bone turnover, caused from direct stimulation of bone cells by increased thyroid hormones. In this study, we aimed to investigate serum osteocalcin levels as a bone formation marker, before antithyroid (propylthiouracil) therapy at hyperthyroid stage and after antithyroid therapy at euthyroid stage of the patients. Twenty four hyperthyroid patients (18 females, 6 males) and 20 (13 females, 7 males) healthy controls were included into this study. Blood and urine samples were taken before medical treatment at hyperthyroid state, and after the antithyroid therapy until the patients reached the euthyroid state. Serum alkaline phosphatase, osteocalcin, calcium, phosphorus, Free T3, Free T4, TSH and urine calcium/creatinine levels were assessed. We found a significant decrease in serum osteocalcin (p=0.006), urinary calcium/creatinine (p=0.004), and serum phosphorus (p=0.038) levels in euthyroid state in comparison to hyperthyroid state. The increases in serum bone formation marker osteocalcin and bone resorption marker urinary calcium/creatinine levels in hyperthyroid state compared to euthyroid state in our study confirmed that hyperthyroid patients have high bone turnover. We conclude that, hyperthyroid patients has high bone turnover of formation and resorption even after attainment of euthyroidism. Osteocalcin and urine calcium/creatinine are sensitive markers in documenting bone remodeling during treatment of hyperthyroidism.


Further characterization of a model of chronic endotoxemia in the rat: adenine nucleotide content in liver.
Male Sprague-Dawley rats (320-380 g) were treated with Escherichia coli endotoxin (ET) either acutely (3 and 4.5 mg/100 g b.w.,) or chronically (0.1 mg/100 g b.w./24 h, i.v. from subcutaneously implanted osmotic pumps). Control rats received only sterile saline. At 6 h after injection and after 30, 54, 126, and 198 h of continuous ET infusion, the animals were sacrificed, and adenine nucleotide content in liver was assayed. Acute ET treatment with either dose produced a slight decrease in ATP content (15% and 11% with 3 mg and 4.5 mg ET/100 g b.w., respectively) associated with an increase of ADP and AMP content and a fall of the energy charge (by 9.2% and 6.4%, with the two ET doses, respectively). Chronic ET treatment did not affect either of the measured parameters. However, in both saline- and ET-infused rats a decrease of ATP content paralleled by an increased level of ADP and AMP and by a fall of energy charge were seen when compared to acutely saline-injected rats. Such changes can be attributed to the surgical trauma associated with pump implantation. It is concluded that the energy status of the liver during endotoxemia cannot account for metabolic disturbances associated with this pathophysiological state.


Phylogeography and population genetics of the endemic Malagasy bat, Macronycteris commersoni s.s. (Chiroptera: Hipposideridae).
Macronycteris commersoni (Hipposideridae), a bat species endemic to Madagascar, is widespread across the island and utilizes a range of habitat types including open woodland, degraded habitats, and forested areas from sea level to 1,325 m. Despite being widely distributed, there is evidence that M. commersoni exhibits morphological and bioacoustic variation across its geographical range. We investigated the fine-scale phylogeographic structure of populations in the western half of the island using extensive spatial sampling and sequence data from two mitochondrial DNA regions. Our results indicated several lineages within M. commersoni. Individuals collected from northern Madagascar formed a single monophyletic clade (clade C). A second clade (clade B) included individuals collected from the south-western portion of the island. This second clade displayed more phylogeographical partitioning with differences in mtDNA haplotypes frequency detected between populations collected in different bioclimatic regions. Lineage dispersal, genetic divergence, and timing of expansion events of M. commersoni were probably associated with Pleistocene climate fluctuations. Our data suggest that the northern and the central western regions of Madagascar may have acted as refugia for this species during periods of cooler and drier climate conditions associated with the Pleistocene.


Design and feasibility of quick EXAFS scans for a 'broomstick' double-crystal monochromator at PLS beamline.
A data-collection technique for quick extended X-ray absorption fine-structure spectroscopy (QEXAFS) was developed with a new 'broomstick' double-crystal monochromator, which has been installed for X-ray absorption fine-structure (XAFS) applications at the Pohang Light Source. The monochromator operates in a fixed-exit scan mode as the Bragg angle is varied from 8 to 80 degrees, corresponding to 2-14 keV, using an Si(111) crystal. The monochromator scan capability was investigated by analysing EXAFS data quality from step-scan and from continuous rotation of the Bragg crystal reflection angle. In our fast continuous-scan design, the electronic pulsing speed of the step motor is adjustable to avoid the monochromatic beam instability caused by serious mechanical resonance. The feasibility of QEXAFS scanning is demonstrated by a typical EXAFS scan (e.g. 1 keV range) being taken within 1 min.


A Pulmonary Embolism Response Team: initial experiences and future directions.
Acute pulmonary embolism (PE) is a common cardiovascular condition resulting in significant morbidity and mortality. Consensus recommendations suggest risk stratification of patients into three main categories: high-risk or 'massive' PE, intermediate-risk or 'submassive' PE, and low-risk PE. Given the relative dearth of prospective, randomized clinical trials delineating optimal selection of the diverse medical, interventional, and surgical treatment approaches, clinical care requires a multidisciplinary expert approach to patients with PE. Areas covered: The Massachusetts General Hospital (MGH) Pulmonary Embolism Response Team (PERT) was the first of its kind to create a multidisciplinary, rapid response team for acute PE, integrated within a research and educational framework. The MGH PERT has treated more than 700 patients with PE, the majority of which are in the 'massive' or 'submassive' categories. The PERT Consortium™ was founded in 2015 as a collaborative network between the growing number of PERT programs internationally, with greater than 80 institutions participating within one year of establishment. Expert commentary: Since its advent, the PERT model has expanded throughout the United States and internationally through a collaborative institutional and research network. PERT may represent a new standard for the care of patients with acute PE.


[Carotid artery wall changes in young type-1 diabetics. The ultrasonic diagnosis of early arteriosclerosis].
In a prospective study high-resolution ultrasonography was used to document any early carotid-artery wall changes (intima-media thickening; plaques), in 165 type I diabetics up to the age of 40 years (66 males, 99 females; mean age 27.5 +/- 6 years; duration of diabetes > 1 year). In two control groups (group 1: 40 with type I diabetes of up to one year's duration; group 2: 20 healthy subjects) the mean intima-media thickness was 0.50 +/- 0.05 and 0.51 +/- 0.06 mm, respectively (maximal value 0.6 mm). In the patients with a longer duration of diabetes it was significantly higher at 0.56 +/- 0.11 mm (P < 0.001). Of the 165 patients with a diabetes duration of more than 1 year, 134 had normal intima-media wall thickness (< 0.6 mm, mean 0.52 +/- 0.07 [group A]), in 15 (group B) it was 0.75 +/- 0.06 mm, and in 16 (group C) there were plaques with a wall thickness of 0.67 +/- 0.15 mm. The incidence of nephropathy and hypertension or hypercholesterolaemia (only in group C) was significantly higher in groups B and C than A (P < 0.01). It is thus clear that these consequential or associated diseases go together, in young type I diabetics, with an increased risk of early arteriosclerosis.


Do parental questions and topic continuations elicit replies from developmentally delayed children? A sequential analysis.
This sequential analysis tested the relative extent to which several adult utterance types elicited conversational replies from developmentally delayed children. Eight developmentally delayed children in Brown's stages I and II and their primary parents were the subjects. Parent-child pairs were video and audio taped during their interactions with experimenter-provided toys in a lab setting. Transcripts of the interactions were coded for adult topic relatedness and obligation level and for child topic relatedness, length, and intelligibility. The results indicated that child replies of any length were elicited by adult topic continuations more than by any other adult utterance type. If a new topic was initiated, explicit prompts for child talk elicited child replies more than other adult utterance types. Multiword child replies were most likely to be elicited by explicit prompts that continued the child's topic. Child effects on the presence and effectiveness of adult conversational recruiting strategies were also tested.


[Breaking bad news in medicine: strategies that turn necessity into a virtue].
Breaking bad news is one of physician's most difficult duties. In spite of this, communication skills are considered a minor professional ability. Bad news is defined as any news that drastically and negatively alters the patient's view of his/her future. Patients never forget where, when and what bad news they were told. The factors that make communicating bad new difficult may come from the sociocultural setting of the patient or the doctor. Randomized studies have shown that communication learning skills may cause a positive impact on the person giving and receiving the bad news. In order to facilitate this task, a practical and dynamic 6-step protocol, described by Baile and Buckman, that may be useful for this task, is described.


Evaluation of exome variants using the Ion Proton Platform to sequence error-prone regions.
The Ion Proton sequencer from Thermo Fisher accurately determines sequence variants from target regions with a rapid turnaround time at a low cost. However, misleading variant-calling errors can occur. We performed a systematic evaluation and manual curation of read-level alignments for the 675 ultrarare variants reported by the Ion Proton sequencer from 27 whole-exome sequencing data but that are not present in either the 1000 Genomes Project and the Exome Aggregation Consortium. We classified positive variant calls into 393 highly likely false positives, 126 likely false positives, and 156 likely true positives, which comprised 58.2%, 18.7%, and 23.1% of the variants, respectively. We identified four distinct error patterns of variant calling that may be bioinformatically corrected when using different strategies: simplicity region, SNV cluster, peripheral sequence read, and base inversion. Local de novo assembly successfully corrected 201 (38.7%) of the 519 highly likely or likely false positives. We also demonstrate that the two sequencing kits from Thermo Fisher (the Ion PI Sequencing 200 kit V3 and the Ion PI Hi-Q kit) exhibit different error profiles across different error types. A refined calling algorithm with better polymerase may improve the performance of the Ion Proton sequencing platform.


Plastic Surgery Training Worldwide: Part 1. The United States and Europe.
Major differences exist in residency training, and the structure and quality of residency programs differ between different countries and teaching centers. It is of vital importance that a better understanding of the similarities and differences in plastic surgery training be ascertained as a means of initiating constructive discussion and commentary among training programs worldwide. In this study, the authors provide an overview of plastic surgery training in the United States and Europe. A survey was sent to select surgeons in 10 European countries that were deemed to be regular contributors to the plastic surgery literature. The questions focused on pathway to plastic surgery residency, length of training, required pretraining experience, training scheme, research opportunities, and examinations during and after plastic surgery residency. Plastic surgery residency training programs in the United States differ from the various (selected) countries in Europe and are described in detail. Plastic surgery education is vastly different between the United States and Europe, and even within Europe, training programs remain heterogeneous. Standardization of curricula across the different countries would improve the interaction of different centers and facilitate the exchange of vital information for quality control and future improvements. The unique characteristics of the various training programs potentially provide a basis from which to learn and to gain from one another.


Novel HCN sorbents based on layered double hydroxides: sorption mechanism and performance.
Layered double hydroxides (LDHs) and layered double oxides (LDOs) have been prepared and used as sorbents for hydrogen cyanide (HCN). Based on results from sorbent optimization experiments, the optimal performance for HCN removal was found in Ni-Al LDH. As evidenced by fixed-bed sorption studies, the Ni-Al LDO with the greatest surface area showed better performance and outperformed products calcined at 200, 400, or 500 °C, whereas, the Ni-Al LDH showed a more twofold higher sorption capacity than the Ni-Al LDO. Investigation of the mechanisms between HCN and sorbents reveals that the HCN removal by the Ni-Al LDH and Ni-Al LDO leads to the formation of the complex anion, [Ni(CN)4](2-). Nevertheless, the [Ni(CN)4](2-) can enter interlayer region of the Ni-Al LDH due to its anion exchangeability, which endows this LDH with more binding sites, not only on its external surfaces, but also on its internal surfaces located in the interlayer region. In contrast, [Ni(CN)4](2-) were only adsorbed on the external surface of the Ni-Al LDO. As a result, the sorption capacity of the Ni-Al LDH for HCN is twice as high as that of the Ni-Al LDO, which is at 21.55 mg/g.


Developing and Validating Metamodels of a Microsimulation Model of Infant HIV Testing and Screening Strategies Used in a Decision Support Tool for Health Policy Makers.
Background. Metamodels can simplify complex health policy models and yield instantaneous results to inform policy decisions. We investigated the predictive validity of linear regression metamodels used to support a real-time decision-making tool that compares infant HIV testing/screening strategies. Methods. We developed linear regression metamodels of the Cost-Effectiveness of Preventing AIDS Complications Pediatric (CEPAC-P) microsimulation model used to predict life expectancy and lifetime HIV-related costs/person of two infant HIV testing/screening programs in South Africa. Metamodel performance was assessed with cross-validation and Bland-Altman plots, showing between-method differences in predicted outcomes against their means. Predictive validity was determined by the percentage of simulations in which the metamodels accurately predicted the strategy with the greatest net health benefit (NHB) as projected by the CEPAC-P model. We introduced a zone of indifference and investigated the width needed to produce between-method agreement in 95% of the simulations. We also calculated NHB losses from "wrong" decisions by the metamodel. Results. In cross-validation, linear regression metamodels accurately approximated CEPAC-P-projected outcomes. For life expectancy, Bland-Altman plots showed good agreement between CEPAC-P and the metamodel (within 1.1 life-months difference). For costs, 95% of between-method differences were within $65/person. The metamodels predicted the same optimal strategy as the CEPAC-P model in 87.7% of simulations, increasing to 95% with a zone of indifference of 0.24 life-months ( ∼ 7 days). The losses in health benefits due to "wrong" choices by the metamodel were modest (range: 0.0002-1.1 life-months). Conclusions. For this policy question, linear regression metamodels offered sufficient predictive validity for the optimal testing strategy as compared with the CEPAC-P model. Metamodels can simulate different scenarios in real time, based on sets of input parameters that can be depicted in a widely accessible decision-support tool.


UV and storage stability of retinol contained in oil-in-water nanoemulsions.
This study was performed to examine the stability of retinol contained within oil-in-water (O/W) emulsions under UV and during storage at different temperatures. O/W emulsions were prepared using different emulsifiers and oil concentrations. The stability of the retinol contained in the O/W emulsions was investigated by measuring the percentage of residual retinol in the samples after UV exposure and storage at different temperatures (4, 25, and 40 °C). The oil concentration of the emulsion had a greater impact on UV stability than the type of emulsifier used, whereas the storage stability at different temperatures was affected by both the choice of emulsifier and the oil concentration. The storage stability of the retinol contained in the O/W emulsions may be related to the lipid oxidation properties of the emulsions rather than the latter's physical stability. Experiments with EDTA and different oil types were performed to confirm this theory.


Background progenitor activation is associated with recurrence after hepatectomy of combined hepatocellular-cholangiocarcinoma.
Hepatic progenitor cells (HPC) play important roles in both liver regeneration and carcinogenesis. Combined hepatocellular-cholangiocarcinoma (CHC), a malignant primary liver tumor with poor prognosis, is thought to be of HPC origin. However, the prognostic significance of this etiology is not well defined. Therefore, we retrospectively investigated the relationship of HPC-related pathological features and long-term outcome in patients with CHC in our department. In a cohort of 80 patients identified between 1997 and 2003, including 70 patients who underwent resection with curative intent, overall survival (OS) and disease-free survival (DFS) were correlated with the proliferative activity of nontumor ductular reaction (DR) and the expression levels of HPC and biliary markers including α-fetoprotein (AFP), keratin 7 (K7), keratin 19 (K19), oval cell (OV)-6, epithelial cell adhesion molecule (EpCAM), and c-Kit in both tumor and nontumor liver. We found that nontumor ductular reactions (DRs), specifically the proliferating cell nuclear antigen (PCNA) labeling index of the ductular reaction (PI-DR), a surrogate for transit-amplifying compartments, was an independent prognostic factor for both OS and DFS. By contrast, intratumoral expression of only one marker, absence of AFP, was associated with OS. PI-DR was also independently associated with synchronous "multicentric occurrence" in hepatocellular carcinoma components, a feature of CHC that may predispose to metachronous multifocal tumorigenesis. Proliferative ductular reaction related to HPC activation is associated with recurrence of CHC. Background HPC activation is strongly associated with multifocal occurrence and related tumor recurrence, highlighting the critical role of background liver disease, a "field effect," in the recurrence of CHC.


Electrogastrography: poor correlation with antro-duodenal manometry and doubtful clinical usefulness in adults.
To investigate if there is a correlation between electrical activity measured by electrogastrography (EGG) and contractile activity of the stomach as measured by antroduodenal manometry (ADM). We also studied whether the underlying motility disorder could be predicted from EGG parameters. We compared 21 parameters measured from EGG with 8 parameters measured from ADM. The ability of EGG to identify the underlying diagnosis was tested by comparing EGG parameters for each diagnosis group against other patients. The study comprised recordings from 148 patients and 125 females. Their median age was 45 (range 17-76) years. We found few and weak correlations between EGG and ADM. Specifically the correlation between parameters reflecting the response to meal was poor (r=-0.07, P=0.39). The discriminatory power of EGG for underlying motility disorder was also low. Patients with slow transit constipation (STC) showed a lower postprandial power in normogastric (3.7+/-0.5 vs 4.0+/-0.5) and tachygastric (3.5+/-0.4 vs 3.7+/-0.4) regions, a lower percentage of time with normogastria [87.2 (56.5-100)% vs 95.7 (0-100)%], and a higher percentage of time with tachygastria [9.3 (0-33)% vs 3.5 (0-100)%] and bradygastria [1.8 (0-20)% vs 0 (0-17.1)%]. Patients with irritable bowel syndrome had a higher percentage of time with normogastria [96.5 (62.5-100)% vs 93.3 (0-100)%] and a less unstable dominant frequency as measured by the instability coefficient [15 (3-77) vs 24 (2-72)]. EGG and ADM seem to measure different aspects of gastric motor activity but cannot show a spatial correlation. The diagnostic value of EGG is poor, but EGG may have some value for the identification of patients with STC.


Partially involuting congenital hemangioma with pyogenic granuloma-induced rapid progression following incisional biopsy in infancy: A case report.
We present a case of partially involuting congenital hemangioma (PICH) that showed rapid growth with ulceration after incisional biopsy. PICH does not typically grow after birth. However, if growth occurs due to stimuli like biopsy, it is essential to consider the possibility of pyogenic granuloma complication.


Approaches on Ferrofluid Synthesis and Applications: Current Status and Future Perspectives.
Ferrofluids are colloidal suspensions of iron oxide nanoparticles (IONPs) within aqueous or nonaqueous liquids that exhibit strong magnetic properties. These magnetic properties allow ferrofluids to be manipulated and controlled when exposed to magnetic fields. This review aims to provide the current scope and research opportunities regarding the methods of synthesis of nanoparticles, surfactants, and carrier liquids for ferrofluid production, along with the rheology and applications of ferrofluids within the fields of medicine, water treatment, and mechanical engineering. A ferrofluid is composed of IONPs, a surfactant that coats the magnetic IONPs to prevent agglomeration, and a carrier liquid that suspends the IONPs. Coprecipitation and thermal decomposition are the main methods used for the synthesis of IONPs. Despite the fact that thermal decomposition provides precise control on the nanoparticle size, coprecipitation is the most used method, even when the oxidation of iron can occur. This oxidation alters the ratio of maghemite/magnetite, influencing the magnetic properties of ferrofluids. Strategies to overcome iron oxidation have been proposed, such as the use of an inert atmosphere, adjusting the Fe(II) and Fe(III) ratio to 1:2, and the exploration of other metals with the oxidation state +2. Surfactants and carrier liquids are chosen according to the ferrofluid application to ensure stability. Hence, a compatible carrier liquid (polar or nonpolar) is selected, and then, a surfactant, mainly a polymer, is embedded in the IONPs, providing a steric barrier. Due to the variety of surfactants and carrier liquids, the rheological properties of ferrofluids are an important response variable evaluated when synthesizing ferrofluids. There are many reported applications of ferrofluids, including biosensing, medical imaging, medicinal therapy, magnetic nanoemulsions, and magnetic impedance. Other applications include water treatment, energy harvesting and transfer, and vibration control. To progress from synthesis to applications, research is still ongoing to ensure control of the ferrofluids' properties.


[Cardiac arrhythmias in patients with chronic alcoholism during abstinence].
A study was undertaken to examine 25 chronic alcoholics in abstinence. A 24-hour monitoring was performed during and extra this state. Chest x-ray study, bicycle ergometry, ultrasonic study of the heart, transesophageal pacing and blood thyroid hormone levels were applied to exclude other causes of cardiac arrhythmias. In alcoholic abstinence, various cardiac arrhythmias appeared as ventricular premature beats (including those of high grade), supraventricular premature beats, supraventricular tachycardias and paroxysmal atrial fibrillation were recorded in all 25 patients. When the abstinence was absent, no cardiac arrhythmias were found in 11 (44%) patients, in 14 (56%) patients who had various arrhythmias, their frequency and number of attacks significantly being lower.


How Might the Relation of the Development of Hand Preferences to the Development of Cognitive Functions be Examined During Infancy: A Sketch?
Investigations of the relation of the development of hand preferences to the development of other sensorimotor and cognitive abilities are plagued by confusing and contradictory results. In part, the confusion derives from the failure to create accurate, appropriate, and reliable descriptions of the development of hand preferences and the cognitive or sensorimotor ability of interest. This paper sketches an ideal longitudinal study (from birth through 5 years) with a large sample size that should provide reliable evidence for the understanding of the relation of hand preferences to cognitive development. Since hand preference differences would affect the way infants engage in manual actions with objects and these differences would likely affect how they come to comprehend object relations, differences in the development of cognition across handedness groups would be a good test of certain forms of embodiment theory.


Animal models and brain circuits in drug addiction.
Animal models in the field of addiction are considered to be among the best available models of neuropsychiatric disease. These models have undergone a number of refinements that allow deeper understanding of the circuitry involved in initiating drug seeking and relapse. Notably, the demonstrable involvement of classic corticostriatal habit circuitry and the engagement of prefrontal cortical circuits in extinction training may have relevance to the therapeutic modulation of habit circuitry and drug addiction in humans.


Improved extremal optimization for the Ising spin glass.
A version of the extremal optimization (EO) algorithm introduced by Boettcher and Percus is tested on two- and three-dimensional spin glasses with Gaussian disorder. EO preferentially flips spins that are locally "unfit"; the variant introduced here reduces the probability of flipping previously selected spins. Relative to EO, this adaptive algorithm finds exact ground states with a speedup of order 10(4) (10(2) ) for 16(2) - (8(3) -) spin samples. This speedup increases rapidly with system size, making this heuristic a useful tool in the study of materials with quenched disorder.


Phytochemicals of Rhus spp. as Potential Inhibitors of the SARS-CoV-2 Main Protease: Molecular Docking and Drug-Likeness Study.
The outbreak of coronavirus disease 2019 (COVID-19) induced by the novel coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) originated in China and spread to cover the entire world with an ongoing pandemic. The magnitude of the situation and the fast spread of the new and deadly virus, as well as the lack of specific treatment, led to a focus on research to discover new therapeutic agents. In this study, we explore the potential inhibitory effects of some active polyphenolic constituents of Rhus spp. (sumac) against the SARS-CoV-2 main protease enzyme (Mpro; 6LU7). 26 active polyphenolic compounds of Rhus spp. were studied for their antiviral activity by molecular docking, drug likeness, and synthetic accessibility score (SAS) as inhibitors against the SARS-CoV-2 Mpro. The results show that all tested compounds of sumac provided good interaction with the main active site of SARS-CoV-2 Mpro, with better, lower molecular docking energy (kcal/mol) compared to the well-known drugs chloroquine and favipiravir (Avigan). Only six active polyphenolic compounds of Rhus spp. (sumac), methyl 3,4,5-trihydroxybenzoate, (Z)-1-(2,4-dihydroxyphenyl)-3-(3,4-dihydroxyphenyl)-2-hydroxyprop-2-en-1-one, (Z)-2-(3,4-dihydroxybenzylidene)-6-hydroxybenzofuran-3(2H)-one, 3,5,7-trihydroxy-2-(4-hydroxyphenyl)chroman-4-one, 2-(3,4-dihydroxyphenyl)-3,5-dihydroxy-7-methoxy-4H-chroman-4-one, and 3,7-dihydroxy-2-(4-hydroxyphenyl)chroman-4-one, were proposed by drug likeness, solubility in water, and SAS analysis as potential inhibitors of Mpro that may be used for the treatment of COVID-19. Six phenolic compounds of Rhus spp. are proposed for synthesis as potential inhibitors against Mpro and have potential for the treatment of COVID-19. These results encourage further in vitro and in vivo investigations of the proposed ligands and research on the preventive use of Rhus spp. against SARS-CoV-2.


In vitro complementation of xeroderma pigmentosum.
In vitro complementation is a powerful strategy for isolation and characterization of individual components of multienzyme biochemical pathways. Application of the method to elucidate DNA metabolic pathways in prokaryotes enabled the successful identification of pathways of DNA replication and repair. In practice the technique requires the availability of genetic mutants that display defective operation of a selected biochemical pathway, and an in vitro assay system that allows detection of effective operation of the pathway. Xeroderma pigmentosum is a human disease syndrome characterized by partial or severe deficit in the operations of the nucleotidyl DNA excision repair pathway. This pathway of DNA repair appears to respond to DNA lesions which produce substantial distortion of helical structure, the best characterized of which are the UV radiation-induced pyrimidine dimers. This review summarizes a variety of approaches to analysis of the reparative deficiencies in xeroderma pigmentosum by in vitro complementation.


Exposure Effects on the Productivity of Commercial Bombus impatiens (Hymenoptera: Apidae) Quads During Bloom in Watermelon Fields.
In light of population declines of honey bees (Apis mellifera L.), research has refocused attention on alternative pollinators and their potential to fulfill pollination services within economically important agricultural crops. Bumble bees are one such alternative, and within the past 20 yr, these pollinators have been reared and sold as commercial pollinators. Investigation into their use has been limited and more research is needed to improve pollinator effectiveness in field settings. Quad pollination units of the commercially reared native bumble bee species, the common eastern bumble bee (Bombus impatiens Cresson), were monitored and evaluated for productivity during peak watermelon [Citrullus lanatus (Thunberg) Matsumura & Nakai] bloom in southern Delaware. Differing colony exposures including various shade structure designs and natural shade were compared to assess the quality of the shade in regards to bumble bee activity during watermelon bloom. Quads receiving different nest treatments were evaluated on the basis of foraging activity and colony weight gain. Results indicated that colonies within quads provided with artificial or natural shade had significantly more foraging activity, weighed more, and produced more cells than colonies in quads placed in the field with no shade. Colonies within quads provided with artificial and natural shade peaked later in terms of foraging and weight gain, suggesting that growers could extend harvest to take advantage of later markets and possible movement into fields that were planted later.


Replicable and shape-controllable fabrication of electrospun fibrous scaffolds for tissue engineering.
Controlling the architecture of electrospun fibers is one of the key issues in tissue engineering. This report describes a rapid and reproducible patterning method for the fabrication of an electrospun fibrous scaffold. The electrospun fibers were deposited on a patterned electrode. The patterned scaffold was fabricated using a thin insulating film between layers of this electrode. For a tissue engineering application, poly(lactic-co-glycolic acid) (PLGA), a biocompatible and biodegradable material, was electrospun. Fibroblast cells were cultured on the fibrous PLGA scaffold and the viability, morphology, and distribution of the cells were studied.


Imbalance between matrix metalloproteinases and their tissue inhibitors in preeclampsia and gestational trophoblastic diseases.
The invasion cascade exhibited by placental trophoblasts and cancerous cells bears many similarities, and it is attributed to extracellular matrix degradation mediated by matrix metalloproteinases (MMPs). Although proper and controlled invasion by trophoblasts into the maternal uterus is an essential requirement for maintenance of normal pregnancy, any abnormality in this phenomenon results in the development of invasion-related disorders such as gestational trophoblastic diseases (GTDs) and preeclampsia. We studied the epigenetic basis of differential expression of two placental MMPs (MMP2 and MMP9) and tissue inhibitors of metalloproteinases (TIMP2 and TIMP1) during normal gestation and invasion-related disorders, i.e., preeclampsia and GTDs. Our study suggests the association of H3K9/27me3 with differential expression of these MMPs and their inhibitors, which regulate the placental invasion during normal pregnancy, whereas no role of CpG methylation was observed in the differential expression of MMPs/TIMPs. Further, development of GTDs was associated with abnormally higher expression of these MMPs and lower levels of their inhibitors, whereas the reverse trends were observed for MMPs and their TIMPs in case of preeclampsia, in association with abnormal changes in H3K9/27me3. These results suggest the involvement of higher levels of MMPs in an aggressive invasive behavior depicted by GTDs, whereas lower levels of these MMPs in shallow and poor invasive phenotype associated with preeclampsia. Thus, our study shows the significance of a proper balance regulated by histone trimethylation between differential expression of MMPs and their TIMPs for maintaining normal pregnancy and its deregulation as a contributing factor for pathogenesis of invasive disorders during pregnancy.


Restoration of caveolin-1 expression suppresses growth, membrane-type-4 metalloproteinase expression and metastasis-associated activities in colon cancer cells.
Caveolin-1 (cav-1) and flotillin-1 are two major structural proteins associated with lipid rafts in mammalian cells. The membrane-type matrix metalloproteinases (MT-MMPs) are expressed at the cell surface, hydrolyze extracellular matrix, and play an important role in cancer cell migration and metastasis. Expression of cav-1, flotillin-1, and MT4-MMP in lysates and lipid rafts of LS174T and HM-7 colon cancer cells was determined. The impact of restoration of cav-1 expression on proliferation, adhesion, motility in vitro, and growth of implanted tumors in vivo was characterized. Cav-1 is not expressed in lipid rafts of the highly metastatic colon cancer cell line (HM-7), but expressed in cytosolic fractions of the parental lower metastatic cell line (LS174T). In contrast, MT4-MMP was expressed in lipid rafts of HM-7 cells but not in LS174T cells. Overexpression of cav-1 in HM-7 cells down-regulate proliferation, viability, wound closure, adhesion to laminin, invasion, and development of filopodial and lamellipodial structures in a dose-dependent manner. Cav-1 positive HM-7 clones ceased to express MT4-MMP in their lipid rafts. Comparative proteomic analyses of lipid rafts from cav-1 positive and cav-1 negative cells demonstrated de novo expression of flotillin-1 only on the cells expressing cav-1. Xenografting control cells devoid of cav-1 in nude mice induced development of bigger tumors expressing higher levels of proliferating cell nuclear antigen as compared to mice injected with cells expressing the highest cav-1 levels. We conclude that cav-1 orchestrates and reorganize several proteins in lipid rafts, activities directly associated with reduced tumorigenic and metastatic ability of colon cancer cells.


Impact of the COVID-19-induced lockdown on the incidence of ocular trauma presenting to a tertiary care hospital.
To assess the effects of lockdown and unlock phases mandated in view of COVID-19 on the incidence and characteristics of ocular trauma presenting to a tertiary care hospital. The study was carried out as a hospital record based retrospective comparative analysis on patients presenting with ocular trauma in the lockdown period (March-July 2020) compared with the same time frame of the previous year considered as prelockdown period (March-July 2019) and during the unlock phases (August-December 2020). Overall, the casualty department saw 464 patients of ocular trauma in the prelockdown period, 173 in the lockdown and 253 in unlock. The study showed a 44% reduction in patients visiting the casualty department for trauma during the lockdown compared with prelockdown, and a 62% reduction specifically in ocular trauma. The unlock phase showed a 21% reduction in ocular trauma compared with prelockdown and a 41% increase compared with the lockdown. In all three phases, the majority of people affected by ocular trauma were middle aged males from a rural background, sustained by assault. The lockdown saw a decrease in outdoor assaults (45%) and road traffic accidents (22%). Trauma sustained by females (18%) increased in the lockdown, as did home-based assaults (150%) and sexual assaults. The presentation of trauma, especially road traffic accidents and outdoor assaults saw a steady rise during the unlock. The lockdown mandated by the government in response to the COVID-19 pandemic had a significant impact on the trends of trauma presenting to healthcare facilities. There was a decrease in the overall number of patients approaching the casualty during the lockdown. However, during the lockdown, there was an increase in home-based trauma as opposed to outdoor assaults being the primary cause of trauma prior to the lockdown.


Pacing for the prevention of atrial fibrillation.
Multiple randomized clinical trials have demonstrated that atrial or dual-chamber pacing prevents paroxysmal and permanent atrial fibrillation (AF) in patients with symptomatic bradycardia as the primary indication for cardiac pacing. The benefit of atrial pacing for the prevention of AF is observed predominantly in patients with sinus node dysfunction. Emerging evidence also suggests that the risk of AF is directly linked to the proportion of time that ventricular pacing occurs. Consequently, pacemakers should be programmed to minimize the amount of ventricular pacing in patients with intrinsic atrioventricular conduction. Temporary atrial pacing following heart surgery may be of benefit for the prevention of perioperative AF. Atrial pacing has not been shown to prevent AF in patients without symptomatic bradycardia. In addition, selective pacing algorithms designed to prevent AF have minimal or no incremental benefits for the prevention of AF. At present, the role of selective atrial lead site(s) for the prevention of AF remains uncertain.


Recombinant adenovirus-p53 (Gendicine) sensitizes a pancreatic carcinoma cell line to radiation.
In this study, we examine the effects of recombinant adenovirus-p53 (rAd-p53) on the pancreatic carcinoma cell line SW1990. Specifically, we determine if expression of rAd-p53 sensitizes these cells to radiation. Following transfection of SW1990 cells with rAd-p53, we measured expression of P53, P21 and Bax by immunocytochemistry. Both transfected and control cell lines were irradiated with a range of doses, and the survival fractions (SF) were calculated. Dose survival curves were constructed and modeled for comparison. Transfection of SW1990 cells with rAd-p53 resulted in increased expression of P53, P21 and Bax in a time-dependent manner. At 96 h after transfection, 89.92% of cells expressed P53, 56.8% expressed P21, and 76.50% expressed Bax. The SF following radiation was lower in the rAd-p53 transfected cells compared to the control cells, suggesting that rAd-p53 sensitizes SW1990 cells to radiation (D0 for the experimental and control groups was 2.199 and 2.462, respectively). Use of the adenoviral vector is an effective means of transfecting SW1990 cells with wild-type P53, and this sensitizes the cell line to irradiation. This work suggests that combining rAd-p53 with radiation therapy in pancreatic cancer may be therapeutically beneficial.


Identifying Cell Cycle Modulators That Selectively Target ARID1A Deficiency Using High-Throughput Image-Based Screening.
ARID1A, a component of the chromatin remodeling complex SWI/SNF, is an evolutionarily conserved complex that uses the energy of adenosine triphosphate hydrolysis to remodel chromatin structure and functions as a master regulator of gene transcription. Recent genomic studies have revealed that ARID1A is one of the most frequently mutated genes in human cancers. However, therapeutic approaches that selectively target ARID1A-mutant tumors are not yet clinically available. Our previous study showed that ARID1A facilitates chromatin response and cell cycle checkpoint activation after DNA damage. Therefore, an ARID1A deficiency may result in therapeutic vulnerabilities in cell cycle modulators. The goals of our study were to develop a novel screening approach, based on fluorescent ubiquitination-based cell cycle indicators (FUCCI), and to identify chemical agents that can selectively modulate the cell cycle transition in ARID1A-deficient cancer cells. Using this high-throughput assay, we screened 2643 compounds and identified six potential chemical modulators that can selectively modulate the cell cycle in ARID1A-deficient cells; these agents may be useful for developing new therapeutics for ARID1A-mutant tumors. In summary, our study demonstrates that FUCCI cell-based high-content screening is a powerful and effective approach for identifying cell cycle modulators and can be applied to multigenotypic screening for targeted cancer therapeutics.


Effects of endocrine disruptors on Bombina orientalis P450 aromatase activity.
To assess the effects of endocrine-disrupting chemicals on the expression and activity of aromatase in the gonads of Bombina orientalis, a common amphibian, we intraperitoneally injected nonylphenol or bisphenol-A and then examined aromatase mRNA levels by RT-PCR as well as aromatase enzymatic activity by tritiated water release assays. To design primers for the RT-PCR, we cloned the B. orientalis aromatase gene using RT-PCR and degenerate primers. The full-length cDNA was obtained by 5'- and 3'-RACE PCR. The complete sequence of the B. orientalis aromatase gene revealed an open reading frame of 1500 bp encoding a deduced protein of 500 amino acids. Semi-quantitative RT-PCR indicated that nonylphenol or bisphenol-A injection did not significantly affect the expression of B. orientalis aromatase mRNA. However, a 48-hr treatment with nonyphenol or bisphenol A reduced aromatase activity to 47% and 32% of the control, respectively. These results suggest that endocrine disrupters can effectively modulate the activity of B. orientalis aromatase without affecting its mRNA levels.


Rich cell-type-specific network topology in neocortical microcircuitry.
Uncovering structural regularities and architectural topologies of cortical circuitry is vital for understanding neural computations. Recently, an experimentally constrained algorithm generated a dense network reconstruction of a ∼0.3-mm3 volume from juvenile rat somatosensory neocortex, comprising ∼31,000 cells and ∼36 million synapses. Using this reconstruction, we found a small-world topology with an average of 2.5 synapses separating any two cells and multiple cell-type-specific wiring features. Amounts of excitatory and inhibitory innervations varied across cells, yet pyramidal neurons maintained relatively constant excitation/inhibition ratios. The circuit contained highly connected hub neurons belonging to a small subset of cell types and forming an interconnected cell-type-specific rich club. Certain three-neuron motifs were overrepresented, matching recent experimental results. Cell-type-specific network properties were even more striking when synaptic strength and sign were considered in generating a functional topology. Our systematic approach enables interpretation of microconnectomics 'big data' and provides several experimentally testable predictions.


The role of sustained posterior brain activity in the serial chaining of two cognitive operations: a MEG study.
A fundamental necessity in human cognition is to link sequential mental operations where appropriate execution of the second task requires input from the first. The present study explores the neural basis of such "chaining" using a novel psychological refractory period (PRP) task. Participants were required to make speeded responses to two sequential visual tasks that were chained or independent. Magnetoencephalography (MEG) signals were recorded simultaneously to reveal the brain's response to these similar but fundamentally different conditions. RTs to Task 1 and 2 were slower in the Chained condition, and their temporal coupling weakened, relative to the Independent condition. MEG analysis of the accompanying event-related fields (ERFs) revealed an increased sustained posterior component in the Chained condition beginning approximately 350 ms after Task 2 onset and lasting for 450 ms. Beamformer localization of this ERF effect revealed a left hemisphere source near the junction of the temporal, parietal, and occipital lobes. These results extend our understanding of the behavioral and corresponding neural mechanisms required by everyday decision making.


Automatic contour extraction from 2D neuron images.
This work describes a novel methodology for automatic contour extraction from 2D images of 3D neurons (e.g. camera lucida images and other types of 2D microscopy). Most contour-based shape analysis methods cannot be used to characterize such cells because of overlaps between neuronal processes. The proposed framework is specifically aimed at the problem of contour following even in presence of multiple overlaps. First, the input image is preprocessed in order to obtain an 8-connected skeleton with one-pixel-wide branches, as well as a set of critical regions (i.e., bifurcations and crossings). Next, for each subtree, the tracking stage iteratively labels all valid pixel of branches, up to a critical region, where it determines the suitable direction to proceed. Finally, the labeled skeleton segments are followed in order to yield the parametric contour of the neuronal shape under analysis. The reported system was successfully tested with respect to several images and the results from a set of three neuron images are presented here, each pertaining to a different class, i.e. alpha, delta and epsilon ganglion cells, containing a total of 34 crossings. The algorithms successfully got across all these overlaps. The method has also been found to exhibit robustness even for images with close parallel segments. The proposed method is robust and may be implemented in an efficient manner. The introduction of this approach should pave the way for more systematic application of contour-based shape analysis methods in neuronal morphology.


News from the Biological Stain Commission.
In the three earlier editions of News from the Biological Stain Commission (BSC), under the heading of "Regulatory affairs," the BSC's International Affairs Committee reported on the work of Technical Committee 212, Clinical Laboratory Testing and in Vitro Diagnostic Test Systems of the International Standards Organization (ISO/TC 212) and its working groups, WG 1, WG 2 and WG 3. In this issue of News from the BSC, H.O. Lyon provides information from the annual meeting of ISO/TC 212 that took place June 2-4, 2008 in Vancouver, British Columbia, Canada. In addition, under the heading of "Certification," J.A. Kiernan examines the certification procedure for thionine used by the BSC laboratory in Rochester, NY.


[Impact of pretreatment variables on the outcome of standardized 131I therapy with 150 Gray in Graves' disease].
We examined the impact of several pretreatment variables on thyroid size and function in 61 patients with Graves' disease one year after a standardized [131]I treatment with 150 Gray. FT3, FT4, and TSH serum concentrations were determined before and 1.5, 3, 6, and 12 months after therapy. Thyroid size was measured by ultrasound and scintigraphy before and one year after therapy. One year after therapy, 30% of the patients had latent or manifest hyperthyroidism, 24% were euthyroid, and 46% had developed latent or manifest hypothyroidism. Age and initial thyroid volume were major predictors of post-therapeutical thyroid function. Thus, persistent hyperthyroidism was observed in 70% of the patients age 50 years and older with a thyroid size of more than 50 ml. With few exception, thyroid size markedly decreased after therapy. Initial thyroid size and age were also major predictors of posttherapeutical thyroid volume. Thyroid size normalized in all patients younger than 50 years of age, independent from initial thyroid size. Radioiodine treatment with 150 Gray causes a considerable decrease in thyroid size in most patients with Graves' disease. Age and initial thyroid volume are important determinants of thyroid function and size after therapy and should be considered in dose calculation.


Isolation of bitter acids from hops (Humulus lupulus L.) using countercurrent chromatography.
Commercially available hops (Humulus lupulus L.) bitter acid extracts contain a mixture of three major congeners (co-, n-, and ad-) in addition to cis/trans diastereomers for each congener. Individual isomerized α-acids were obtained by the consecutive application of two separate countercurrent chromatography methods. First, individual isomerized α-acid congeners as a mixture of cis/trans diastereomers were obtained using a solvent system consisting of hexane and aqueous buffer. The second purification, capable of separating cis/trans diastereomers, was accomplished using a quaternary solvent system; an alternative procedure using β-cyclodextrin followed by countercurrent chromatography was also investigated. The NaBH(4) reduction of the purified isomerized α-acid compounds followed by countercurrent chromatography purification resulted in individual ρ iso α-acids (>95%). Similarly, catalytic hydrogenation of the purified isomerized α-acid compounds followed by countercurrent chromatography purification produced individual tetrahydro isomerized α-acids (>95%). Reported herein is a widely applicable approach that focuses on three critical variables--solvent system composition, pH, and buffer-to-sample ratio--that enable the efficient purification of individual bitter acids (≥95%) from commercially available hops extracts.


Collagen peptides from Acaudina molpadioides prevent CCl4 -induced liver injury via Keap1/Nrf2-ARE, PI3K/AKT, and MAPKs pathways.
Collagen peptide from Acaudina molpadioides (AMP) showed antioxidative activity in H2 O2 -induced RAW264.7 cells in our pervious study. In this study, it was observed that AMP could effectively improve the morphology and function of liver in CCl4 -induced mice. After 200 mg/kg AMP treatment, the content of MDA in liver decreased by 62.3%, and the level of antioxidant enzymes (SOD, GSH-Px, and CAT) increased by more than 65%. Western blot results disclosed that AMP (200 mg/kg) upregulated the Nrf2 level by 73.8% and downregulated Keap1 by 41.0% in CCl4 -induced mice liver. The levels of p-ERK, p-JNK, and p-p38 in 200 mg/kg AMP treatment groups decreased by 57.3%, 40.9%, and 40.6%, but the levels of p-PI3K and p-AKT increased by 162.6% and 60.3%, respectively. Furthermore, the trends of Nrf2, Keap1, p-ERK, p-JNK, p-p38, p-PI3K, and p-AKT levels in H2 O2 -induced RAW264.7 cells after AMP treatment were similar to the results in CCl4 -induced mice liver. These findings provided evidence that AMP exerted antioxidant activity via Keap1/Nrf2-ARE, PI3K/AKT, and MAPKs pathways in vivo and in vitro. Therefore, the collagen peptide from A. molpadioides might represent a novel functional food to prevent acute liver injury via attenuation of oxidative stress.


Nonenzymatic glycosylation of bovine retinal microvessel basement membranes in vitro. Kinetic analysis and inhibition by aspirin.
Incubation of intact bovine retinal microvessels or isolated retinal microvessel basement membranes (RVBM) with radioactive D-glucose or L-glucose, followed by basement membrane collagenous protein purification, resulted in the isolation of nonenzymatically glycosylated RVBM collagens. Type IV collagen was identified in the RVBM by selective salt fractionation, SDS-polyacrylamide gel electrophoresis, amino acid analysis, and immunoprecipitation with specific antibody. Kinetic analysis of the condensation of glucose with RVBM was carried out by labeling retinal microvessel basement membranes with D-[2-3 H]-glucose and D-[6-14 C]-glucose. The rate constant for aldimine product formation, k1, was 1.95 +/- 0.24 (SD) X 10(-4) mM-1 h-1, and the rate constant for the reversed reaction, k-1, was 5.9 +/- 1.0 X 10(-2) h-1. Based on a rate constant for the Amadori rearrangement, k2, of 8.8 +/- 1.0 X 10(-3) h-1, which was the rate-determining step, the half life of this reaction was 80 +/- 9 h. These data may be useful in estimating the glycosylation of retinal microvessel basement membranes in vivo. The nonenzymatic glycosylation of retinal microvessel basement membrane proteins was progressively inhibited by increasing concentrations (0.1 to 2.0 mM) of aspirin.


Mechanism Analysis and Property Prediction of Extended Surfactants Based on the Respectively Optimized Force Field.
Extended surfactants represent a novel class of anionic-nonionic surfactants with exceptional performance and unique application value in chemically enhanced oil recovery. Although molecular dynamics (MD) simulations can efficiently screen these surfactants, the current research is limited. Here, it is proven for the first time that existing generic force fields (GAFF and CHARMM) cannot accurately describe extended surfactants, and traditional approaches are insufficient for obtaining precise charge parameters. The concept of the respectively optimized force field (ROFF) with the purports of specialization and accuracy is proposed to construct high-accuracy models for MD simulations, and a new approach is developed to simulate the interface model. By combining the newly specialized alkane model, ROFF-based surfactant models, and the innovative simulation protocol, high accuracy and reliability can be obtained in predicting hydration free energies, minimum of area per molecule, and critical micelle concentration of extended surfactants. Key properties of the newly designed extended surfactants in conventional oil-water interfaces and oil reservoir environments are comprehensively predicted by using advanced analytical and characterization methods. Furthermore, the more rigorous mechanism underlying the special amphiphilicity of the extended surfactant is revealed, potentially offering significant improvements over previous empirical perspectives.