Pharmacokinetic properties of LMW-heparin-tyramine fractions with high or low affinity to antithrombin III in the rat.
We have recently presented evidence that a macrophage scavenger receptor-mediated pathway is responsible for the hepatic uptake of unfractionated heparins and low-molecular-weight heparins (LMWHs) in the rat. The same receptor-mediated pathway was partially responsible for the removal of oxidized low-density lipoprotein. Unfractionated and fractionated LMWHs exert their anticoagulatory effects predominately by reversibly binding to the plasma glycoprotein antithrombin III. In this study LMWHs modified by endpoint attachment of tyramine were radiolabeled and their fractions with low or high affinity to AT-III studied in vivo in rats. The high-affinity fraction was predominately cleared from the circulation by a hepatic uptake mechanism. About 25% of the injected high-affinity tracer material was recovered, whereas only about 8% of the low-affinity material was found in the liver after 180 minutes. Blocking the scavenger receptor-mediated liver RES uptake mechanism by maleylated bovine serum albumin led to a considerable decline in liver uptake (9 versus 25%). The low-affinity material was rapidly eliminated into the urine after 180 minutes. About 45% of the low-affinity material was excreted versus 23% of the high-affinity material. Tight binding to AT-III prevented LMWH-tyramine from being rapidly cleared from the circulation via the kidneys into the urine; instead, the scavenger receptor-mediated hepatic uptake mechanism seemed to be more dominant in removing material with high affinity to AT-III from blood.


RNAome sequencing delineates the complete RNA landscape.
Standard RNA expression profiling methods rely on enrichment steps for specific RNA classes, thereby not detecting all RNA species. For example, small and large RNAs from the same sample cannot be sequenced in a single sequence run. We designed RNAome sequencing, which is a strand-specific method to determine the expression of small and large RNAs from ribosomal RNA-depleted total RNA in a single sequence run. RNAome sequencing quantitatively preserves all RNA classes. This characteristic allows comparisons between RNA classes, thereby facilitating relationships between different RNA classes. Here, we describe in detail the experimental procedure associated with RNAome sequencing published by Derks and colleagues in RNA Biology (2015) [1]. We also provide the R code for the developed Total Rna Analysis Pipeline (TRAP), an algorithm to analyze RNAome sequencing datasets (deposited at the Gene Expression Omnibus data repository, accession number GSE48084).


Recent advances in the management of osteoporosis.
There has been substantial progress in the management of patients with osteoporosis and the prevention of osteoporotic fractures. Currently available strong anti-resorptive agents are bisphosphonates and an anti-receptor activator of nuclear factor-kappa B ligand (RANKL) antibody, denosumab. Although bisphosphonates and denosumab both inhibit bone resorption and prevent vertebral and non-vertebral fractures, their mechanisms of action are different. Whereas bisphosphonates' effects on bone mineral density and fracture peak around 3 to 5 years and become plateaued, those of denosumab are maintained for up to 10 years. There are differences in the modes of action of these two drugs. Bisphosphonates accumulate on the mineralized bone surface and are released by the acid environment under osteoclastic bone resorption, whereas denosumab is not accumulated on bone but directly binds RANKL and inhibits its binding to the receptor RANK. Thus, the reduction in denosumab concentration 4 to 6 months after injection may enable RANK to bind to RANKL, where it is highly expressed, such as in damaged bone regions. As anabolic agents, only teriparatide has been available for a long time, but abaloparatide, a synthetic analog of PTHrP(1-34), is currently under development. Because of the difference in the preferential binding conformations of PTH1 receptor between teriparatide and abaloparatide, the latter shows anabolic effects with fewer bone resorptive effects. Romosozumab, an anti-sclerostin antibody, inhibits the action of sclerostin, a canonical Wnt signal inhibitor secreted from osteocytes, and enhances canonical Wnt signaling. Romosozumab robustly increases vertebral and proximal femoral bone mineral density within 12 months and inhibits vertebral and clinical fractures in patients with osteoporosis by enhancing bone formation and inhibiting bone resorption. In this review, we summarize the recent advances in therapeutic agents for the treatment of osteoporosis and discuss future prospects with their use.


A privacy-aware method for COVID-19 detection in chest CT images using lightweight deep conventional neural network and blockchain.
With the global spread of the COVID-19 epidemic, a reliable method is required for identifying COVID-19 victims. The biggest issue in detecting the virus is a lack of testing kits that are both reliable and affordable. Due to the virus's rapid dissemination, medical professionals have trouble finding positive patients. However, the next real-life issue is sharing data with hospitals around the world while considering the organizations' privacy concerns. The primary worries for training a global Deep Learning (DL) model are creating a collaborative platform and personal confidentiality. Another challenge is exchanging data with health care institutions while protecting the organizations' confidentiality. The primary concerns for training a universal DL model are creating a collaborative platform and preserving privacy. This paper provides a model that receives a small quantity of data from various sources, like organizations or sections of hospitals, and trains a global DL model utilizing blockchain-based Convolutional Neural Networks (CNNs). In addition, we use the Transfer Learning (TL) technique to initialize layers rather than initialize randomly and discover which layers should be removed before selection. Besides, the blockchain system verifies the data, and the DL method trains the model globally while keeping the institution's confidentiality. Furthermore, we gather the actual and novel COVID-19 patients. Finally, we run extensive experiments utilizing Python and its libraries, such as Scikit-Learn and TensorFlow, to assess the proposed method. We evaluated works using five different datasets, including Boukan Dr. Shahid Gholipour hospital, Tabriz Emam Reza hospital, Mahabad Emam Khomeini hospital, Maragheh Dr.Beheshti hospital, and Miandoab Abbasi hospital datasets, and our technique outperform state-of-the-art methods on average in terms of precision (2.7%), recall (3.1%), F1 (2.9%), and accuracy (2.8%).


Unconjugated payload quantification and DAR characterization of antibody-drug conjugates using high-resolution MS.
The application of high-resolution MS to antibody-drug conjugate (ADC) drug development may provide insight into their safety and efficacy. Quantification of unconjugated cytotoxic drug (payload) and characterization of drug-to-antibody ratio distribution were determined in plasma using orthogonal acceleration quadrupole-time-of-flight MS. Unconjugated payload quantification determined by quadrupole-time-of-flight-based MRM(highresolution) and triple quadrupole-based multiple reaction monitoring were comparable and achieved detection limits of 0.030 and 0.015 ng/ml, respectively. As determined by immunocapture and TOF-MS, drug-to-antibody ratio remained unchanged up to 3-weeks postdose for an ADC containing engineered glutamine linkers, but declined from four to three over 2 weeks in an ADC containing engineered cysteine linkers. The use of high-resolution MS in ADC drug discovery confirms its utility within the bioanalytical discipline.


Investigation into the rejuvenation of spent electroless nickel baths by electrodialysis.
Electroless nickel plating generates substantially more waste than other metal-finishing processes due to the inherent limited bath life and the need for regular bath disposal. Electrodialysis can be used to regenerate electroless nickel baths, but poor membrane permselectivity, leading to high losses of valuable bath components, continues to be a weakness of the technology. This research has investigated improving electrodialysis permselectivity for removing contaminants (sodium, orthophosphite, and sulfate) in a spent electroless nickel bath while minimizing the losses of valuable bath ions (nickel, hypophosphite, and organic acids). Ion permselectivity was explored with respect to electrodialysis operating conditions, membrane type, and cell configuration. Excellent permselectivity for sodium over nickel was attained irrespective of operating condition, membrane, or cell configuration. Studies on the effects of four different operating conditions (current density, pH, flow rate, and temperature) on anion permselectivity revealed bath pH and current density to be critical operating parameters. The type of anion exchange membrane used had a crucial effect on selectivity; one membrane (Ionac MA-3475) was identified as having superior selectivity for bath contaminants particularly for sulfate. The improvements in electrodialysis permselectivity established by this research will decrease waste generation within the electroless nickel process and increase resource productivity by minimizing the loss of valuable plating chemicals.


Fetal perfusion with maternal blood; a possible sequel to fetal catheterization in the pig.
Plasma gamma-globulin concentrations were increased in three catheterized pig fetuses in which the packed red cell volume rose rapidly after surgery, but were not increased in one fetus in which the packed cell volume remained normal. The increases in packed cell volume and gamma-globulin concentration were closely correlated. The source of the gamma-globulin was investigated in one sow with catheterized fetuses. Maternal proteins, identified by [125I]-labelling and gel diffusion, crossed to the circulation of the one catheterized fetus with an elevated packed cell volume but not to the other, in which packed cell volume remained normal, nor to two unoperated fetuses. The results suggest that vascular anastmosis between fetal and maternal circulations is a possible sequel to cannulation in this species.


Haplotypes in CCR5-CCR2, CCL3 and CCL5 are associated with natural resistance to HIV-1 infection in a Colombian cohort.
Variants in genes encoding for HIV-1 co-receptors and their natural ligands have been individually associated to natural resistance to HIV-1 infection. However, the simultaneous presence of these variants has been poorly studied. To evaluate the association of single and multilocus haplotypes in genes coding for the viral co-receptors CCR5 and CCR2, and their ligands CCL3 and CCL5, with resistance or susceptibility to HIV-1 infection. Nine variants in CCR5-CCR2, two SNPs in CCL3 and two in CCL5 were genotyped by PCR-RFLP in 35 seropositive (cases) and 49 HIV-1-exposed seronegative Colombian individuals (controls). Haplotypes were inferred using the Arlequin software, and their frequency in individual or combined loci was compared between cases and controls by the chi-square test. A p' value ;0.05 after Bonferroni correction was considered significant. Homozygosis of the human haplogroup (HH) E was absent in controls and frequent in cases, showing a tendency to susceptibility. The haplotypes C-C and T-T in CCL3 were associated with susceptibility (p'=0.016) and resistance (p';0.0001) to HIV-1 infection, respectively. Finally, in multilocus analysis, the haplotype combinations formed by HHC in CCR5-CCR2, T-T in CCL3 and G-C in CCL5 were associated with resistance (p'=0.006). Our results suggest that specific combinations of variants in genes from the same signaling pathway can define an HIV-1 resistant phenotype. Despite our small sample size, our statistically significant associations suggest strong effects; however, these results should be further validated in larger cohorts.


Identification of active-site inhibitors of MurG using a generalizable, high-throughput glycosyltransferase screen.
MurG is a glycosyltransferase involved in the biosynthesis of bacterial peptidoglycan. It is a potentially important antibiotic target, but no inhibitors of the enzyme have been reported. In general, inhibitors of glycosyltransferases have been difficult to design. Furthermore, no glycosyltransferase inhibitors have been identified through high-throughput screening, perhaps because appropriate screens for glycosyltransferase inhibition have not been developed. In this manuscript, we describe the development of a high-throughput screen for MurG that was used to screen a 50 000 compound library for inhibitors. The screen, which can be generalized to other glycosyltransferases, led to the identification of a family of active-site directed MurG inhibitors. The family of inhibitors contains a five-membered heterocyclic core that appears to function as a diphosphate mimic with respect to the presentation of substituents. We discuss the implications of this result and the utility of the screen for identifying inhibitors of other glycosyltransferases.


[Guideline on optimal blood pressure range for Chinese oldest old].
Hypertension is a major problem of public health that endangers the health of the oldest old. However, the current guidelines for hypertension management do not uniformly diagnose hypertension among the oldest old, nor recommend a normal blood pressure range, which is not convictive enough to support the decision making to the prevention of blood pressure-related adverse events. This guideline gives guiding opinions on optimal blood pressure range for the Chinese oldest old, which applies to the staff of medical and health institutions at all levels nationwide to evaluate the blood pressure levels of the oldest old. It includes the sections of general principles, methods and standards of blood pressure evaluation, measurement conditions, specifications of blood pressure measurement, implementation approaches, etc. The guideline has important directive significance for improving the blood pressure management and decision-making level of the Chinese oldest old.


MHC class I-restricted presentation of maleylated protein binding to scavenger receptors.
Pathways for loading exogenous protein-derived peptides on MHC class I are thought to be present mainly in monocyte-lineage cells and to involve phagocytosis- or macropinocytosis-mediated antigenic leakage into either cytosol or extracellular milieu to give peptide access to MHC class I. We show that maleylation of OVA enhanced its presentation to an OVA-specific MHC class I-restricted T cell line by both macrophages and B cells. This enhanced presentation involved uptake through receptors of scavenger receptor (SR)-like ligand specificity, was TAP-1-independent, and was inhibited by low levels (2 mM) of ammonium chloride. No peptide loading of bystander APCs by maleylated (maleyl) OVA-pulsed macrophages was detected. Demaleylated maleyl-OVA showed enhanced MHC class I-restricted presentation through receptor-mediated uptake and remained highly sensitive to 2 mM ammonium chloride. However, if receptor binding of maleyl-OVA was inhibited by maleylated BSA, the residual presentation was relatively resistant to 2 mM ammonium chloride. Maleyl-OVA directly introduced into the cytosol via osmotic lysis of pinosomes was poorly presented, confirming that receptor-mediated presentation of exogenous maleyl-OVA was unlikely to involve a cytosolic pathway. Demaleylated maleyl-OVA was well presented as a cytosolic Ag, consistent with the dependence of cytosolic processing on protein ubiquitination. Thus, receptor-specific delivery of exogenous protein Ags to APCs can result in enhanced MHC class I-restricted presentation, suggesting that the exogenous pathway of peptide loading for MHC class I may be a constitutive property dependent mainly on the quantity of Ag taken up by APCs.


Screening for cancer.
Screening has proved effective for only two cancer sites, the breast and the cervix uteri. Only for these, therefore, should screening be a part of routine clinical practice. The degree to which screening is effective depends on whether appropriate groups are screened. For cancer of the breast, current evidence justifies mammographic screening every two years only from the age of 50, continuing at least to the age of 70. The place of breast self-examination and physical examination of the breasts is under investigation, but these procedures should probably be used in all women after the age of 40. For cancer of the cervix, nearly maximal efficacy is obtained by screening sexually active women every three years from the age of 25 to 60. Older women who have not been screened regularly should be screened. Other screening techniques to detect cancers, such as the rectal examination, sigmoidoscopy, and fecal occult blood tests for colorectal cancer, are still experimental. Organizational and other difficulties will prevent screening programs from making a major contribution to reducing overall cancer mortality by the year 2000. If organizational problems can be overcome, however, screening may control invasive cancer of the cervix and make a major contribution in reducing breast cancer mortality in the future.


[Screening and identification of stimulant and psychedelic drugs as acetyl-derivatives by GC-MS].
With the increasing number of abused stimulant and psychedelic drugs, along with so called "dance drugs" and "new synthetic drugs" available on the Czech illegal market, there is a need to update methods in toxicological laboratories and therefore it is necessary to develop and optimalize screening and identification procedures for new toxic substances appearing in the laboratory practice. It is well known that relatively popular commercial screening immunoassays have some limits: for instance restricted amount of detectable substances, specificity and sensitivity of detection. Therefore, it is mandatory to combine or complete them with more specific methods based on a different principle. In this paper we have focused on collecting useful analytical data to introduce or complete the system of detection and identification of unknown drugs and their metabolites which can appear in biological samples by using gas chromatography-mass spectrometry (GC-MS) after preparation of relevant acetylated derivatives. The collection of experimental data involves retention indexes and mass spectra of acetylated phenylalkylamines, tryptamines and piperazines and some of their metabolites. These data are fundamental for laboratory diagnostics of drugs of abuse or intoxication and they can be useful for practical application in a number of toxicological laboratories. The mean limit of detection 0.1 ng analyte injected to GC-MS is low enough to allow the method to be successfully applied to real toxicological samples.


Result of percutaneous disc decompression using nucleoplasty in Thailand: a randomized controlled trial.
Chronic low back pain is a major social, economic and healthcare issue in the Thailand. Percutaneous techniques are rapidly replacing traditional open surgery in operations requiring discectomy, decompression and fusion. The percutaneous access to the disc was first used in the 1950s to biopsy the disc with needles. Percutaneous access to the disc using endoscopic techniques was developed in the 1970s. Nucleoplasty has emerged as one of the minimally invasive techniques for treatment of low back pain and lower extremity pain due to contained herniated discs which utilizes coblation technology for ablating and coagulating the nucleus for a partial disc removal. Evaluate the effectiveness of Nucleoplasty on pain in activity and improvement in MRI in patients with radicular or axial low back pain secondary to contained herniated discs. Prospective, Randomized, Control Trial. Sixty-four patients were randomized in two groups equally. Thirty-two patients had undergone Nucleoplasty and another thirty-two patients had undergone conservative treatment. Patients were evaluated at 1, 3, 6 and 12 months postoperatively and were asked to quantify their pain using a visual analog scale ranging from 0 to 10. Data were compared between baselines and at 1, 3, 6 and 12 months post-treatment. Pre-nucleoplasty MRI and Post-nucleoplasty 3 months were compared to evaluate the decrease of bulging disc. Reported pain and medication use were significantly decreased and functional status was improved at 1, 3, 6 and 12 months following Nucleoplasty (p-values < or = 0. 001 for all outcome measures at all time periods) and also the bulging disc was significantly decreased 3 months following nucleoplasty. Nucleoplasty appears to be safe and effective in Thailand. Is an effective procedure for patients presenting with discogenic back and/or radicular pain that have failed conservative therapies and are not considered candidates for open surgical interventions. A result of this analysis indicated that PDD using Coblation technology, also referred to as nucleoplasty, is an effective procedure for patients presenting with discogenic back and/or leg pain who have failed conservative therapies and are not considered candidates for open surgical interventions.


Denaturation/refolding of purified recombinant HIV reverse transcriptase yields monomeric enzyme with high enzymatic activity.
We engineered a prokaryotic expression vector encoding the HIV reverse transcriptase (RT). We grew Escherichia coli JM109 carrying the vector in a 250-liter stirred tank fermentor and purified RT (p66) under native conditions to apparent homogeneity. Purified p66 (greater than or equal to 5 mg/ml) was not stable, and was rapidly processed to its 51 kD derivative (p51), until p66:p51 levels were approximately 1:1. These latter RT preparations were chromatographed as heterodimers and had approximately fivefold higher specific RT enzymatic activities compared with those containing predominantly p66. P66 purified under dilute concentrations (less than or equal to 0.5 mg/ml) was monomeric in solution, resistant to p51 processing for weeks at 4 degrees C, but also had low specific RT enzymatic activities. To attempt the preparation of homogeneous p66 with specific RT enzymatic activities equivalent to p66:p51 heterodimers, purified heterodimers were denatured and p66 was purified and refolded during extensive dialysis (refolded p66). Refolded p66 (less than or equal to 0.5 mg/ml) was monomeric in solution and had identical specific RT enzymatic activities, Km for dTTP, and inhibition by 3'-azido-3'-deoxythymidine triphosphate compared with heterodimeric p66:p51 RT. The data indicates that HIV RT obtained from recombinant E. coli under native conditions is extensively processed at concentrations promoting dimerization. Moreover, RT denaturation and refolding yields apparently homogeneous monomeric p66, with specific RT enzymatic activities equivalent to heterodimeric RT.


Mitochondrial alterations, oxidative stress and neuroinflammation in Alzheimer's disease.
Alzheimer's disease (AD) is a multifactorial disorder characterized by the progressive deterioration of neuronal networks. The primary cause and sequence of its progression are only partially understood but abnormalities in folding and accumulation of insoluble proteins such as beta-amyloid and Tau-protein are both associated with the pathogenesis of AD. Mitochondria play a crucial role in cell survival and death, and changes in mitochondrial structure and/or function are related to many human diseases. Increasing evidence suggests that compromised mitochondrial function contributes to the aging process and thus may increase the risk of AD. Dysfunctional mitochondria contribute to reactive oxygen species which can lead to extensive macromolecule oxidative damage and the progression of amyloid pathology. Oxidative stress and amyloid toxicity leave neurons chemically vulnerable. The mitochondrial toxicity induced by beta-amyloid is still not clear but may include numerous mechanisms, such as the increased permeability of mitochondrial membranes, the disruption of calcium homeostasis, the alteration of oxidative phosphorylation with a consequent overproduction of reactive oxygen species. Other mechanisms have been associated with the pathophysiology of AD. Inflammatory changes are observed in AD brain overall, particularly at the amyloid deposits, which are rich in activated microglia. Once stimulated, the microglia release a wide variety of pro-inflammatory mediators including cytokines, complement components and free radicals, all of which potentially contribute to further neuronal dysfunction and eventually death. Clinically, novel approaches to visualize early neuroinflammation in the human brain are needed to improve the monitoring and control of therapeutic strategies that target inflammatory and other pathological mechanisms. Similarly, there is growing interest in developing agents that modulate mitochondrial function.


Frequent exchange of the DNA polymerase during bacterial chromosome replication.
The replisome is a multiprotein machine that carries out DNA replication. In Escherichia coli, a single pair of replisomes is responsible for duplicating the entire 4.6 Mbp circular chromosome. In vitro studies of reconstituted E. coli replisomes have attributed this remarkable processivity to the high stability of the replisome once assembled on DNA. By examining replisomes in live E. coli with fluorescence microscopy, we found that the Pol III* subassembly frequently disengages from the replisome during DNA synthesis and exchanges with free copies from solution. In contrast, the DnaB helicase associates stably with the replication fork, providing the molecular basis for how the E. coli replisome can maintain high processivity and yet possess the flexibility to bypass obstructions in template DNA. Our data challenges the widely-accepted semi-discontinuous model of chromosomal replication, instead supporting a fully discontinuous mechanism in which synthesis of both leading and lagging strands is frequently interrupted.


Methodological strategies to engage young black and Latino heterosexual couples in sexual and reproductive health research.
Approaches that move beyond individuals and target couples may be an effective strategy for reducing sexual and reproductive health (SRH) disparities among adolescents and young adults (AYA). However, few researchers have attempted to recruit couples due to feasibility and methodological issues. This study aims to enhance implementation and methodological approaches to successfully engage heterosexual Black and Latino adolescent and young adult (AYA) couples in sexual reproductive health (SRH) research. We developed a four-step approach to systematically engage AYA couples in a qualitative study examining factors that influence uptake of combination HIV prevention methods: 1) understanding barriers and facilitators to engaging AYA couples, (2) identifying AYAs living in geographic areas of HIV vulnerability, (3) recruiting and screening AYA couples, and (4) scheduling and completion of the interview session. Black and Latino youth aged 16 to 24 and their opposite sex romantic were recruited in the South Bronx, New York from September 2017-May 2018. Three hundred and seventy-two men and women completed screening procedures to determine eligibility for the index participant; 125 were eligible and enrolled into the study. Forty-nine nominated partners (NPs) participated in screening procedures and enrolled into the study. A total of 49 couples enrolled into the study; 23 couples completed study activities. Developing a systematic recruitment plan aided in successfully engaging Black and Latino heterosexual youth. Nevertheless, barriers to study enrollment remained including locating eligible IPs and screening of the NP. Targeting both young men and women was an effective recruitment strategy. Moreover, dyadic strategies that allow for simultaneous interaction with both couple members may be a beneficial strategy to couples' study enrollment and completion of study activities.


Biochemical and immunochemical studies on the GABAergic system in the rat fallopian tube and ovary.
gamma-Aminobutyric acid (GABA) and glutamic acid decarboxylase (GAD) activities were measured in the ovary and the Fallopian tube of rats and compared with brain values. GABA levels in the Fallopian tube were about twice as high as in the brain, while in the ovary they represented only about 5% of the amino acid content of the CNS. In vitro decarboxylation of glutamate, measured via CO2 formation, occurred both in the Fallopian tube and in the ovary. These two organs contained, respectively, 10% and 1% of brain GAD activity. However, the actual formation of GABA from glutamate in a high-speed supernatant was detectable only in the Fallopian tube, where it represented about 5% of brain GAD activity. In contrast with the enzyme present in ovary, liver, anterior pituitary, and kidney, that in the Fallopian tube was quantitatively precipitated by a specific antiserum directed against rat neuronal GAD. Moreover, subcutaneous transplantation resulted in a quantitative decrease of both GABA levels and GAD activity in the Fallopian tube while no change occurred in the ovary, and vagus nerve section induced a 50% decrease of GAD activity in the Fallopian tube, although GABA levels were not significantly altered. The findings suggest an extrinsic GABAergic innervation in the rat Fallopian tube but not in the ovary.


Cerebral spinal fluid involvement by Hodgkin's disease diagnosed by CSF cytology and immunocytochemistry.
A 39-yr-old man with stage IV Hodgkin's disease (HD) involving bone marrow was being evaluated for autologous bone marrow transplantation when he developed diplopia, prompting a lumbar puncture tap for cerebral spinal fluid (CSF) examination. Cytologic examination of the CSF revealed numerous Reed-Sternberg (RS) cells in a polymorphous inflammatory background of small lymphocytes, monocytes, rare plasma cells, and eosinophils. However, magnetic resonance imaging (MRI) studies of the brain and spinal cord failed to reveal evidence of leptomeningeal disease or intracranial masses. Repeat CSF examination again demonstrated cytologic evidence of HD. Immunocytochemical stains established that the RS cells and mononuclear Hodgkin's cells were positive for CD30 and CD20 but negative for CD15; this phenotype was identical to that of RS cells in the initial diagnostic bone marrow biopsy, confirming CSF involvement by HD. The patient was treated with intrathecal methotrexate, 15 mg, 6 days after his bone marrow transplant. After treatment, all subsequent CSF cytology specimens were negative for tumor. In this case of disseminated HD, cytologic examination allowed for early detection of CNS involvement by lymphoma prior to development of radiographically detectable lesions.


[Quantitative mineralogical analysis and structure of urinary stones in patients living in Ivanovo region].
This paper focuses on developing and implementing a method of quantitative mineralogical analysis of urinary stones based on powder diffraction data analysis using 4 Topas (Bruker) software. Mineralogical composition of 100 urinary stones from urolithiasis patients living in Ivanovo region was examined. More than 70% of stones consisted of calcium oxalate monohydrate (COM) and calcium oxalate dihydrate (COD), and their mixtures with hydroxylapatite. Forty four percent of urinary stones consisted of one component (COM, uric acid (UA) or, less frequently, hydroxyapatite (HA); 56% of urinary stones comprised two, three or four components. The most common mineral was COM (more than 70% of cases), the rarest were calcium oxalate trihydrate (CT), brushite and newberrite. The most common combinations of minerals in mixed stones were COM+HA, COM+COD and COM+COD+HA. The texture, the surface composition and its changes in the course of chemolysis in different types of stones were examined using scanning electron microscopy (SEM) and X-ray microanalysis (XRM). Implications for using analytical chemical and physical techniques for the diagnosis and treatment of urolithiasis were discussed.


Indirect comparisons of treatment effects: Network meta-analyses.
Many therapeutic options are now available for patients with multiple sclerosis. While the efficacy of each drug has been assessed against placebo or, more recently, against interferon, no direct comparisons of these new therapies have been conducted in randomized clinical trials. Therefore, indirect treatment comparisons are needed to inform clinical decisions. In this brief report, some basic concepts about network meta-analyses that are the formal methods used to run multiple indirect treatment comparisons are reviewed when applied in the context of multiple sclerosis studies.


Apical Periodontitis and Cardiovascular Disease in Adults: A Systematic Review with Meta-Analysis.
A potential relationship between oral health and cardiovascular diseases has been proposed. However, uncertainty remains as to whether there is sufficient data to support this association. This review aims to appraise the relationship between apical periodontitis and cardiovascular disease based on data from observational studies. The databases Medline (via PubMed) and EMBASE (via Scopus) were searched up to August 2020 for observational studies (case-control, cross-sectional, and cohort) assessing the association of apical periodontitis with cardiovascular disease among adults. Pooled relative risk/odds ratio and 95% confidence interval (CI) were estimated using a random-effects model. Sensitivity analyses and random-effects meta-regressions were performed. The initial search yielded 2537 documents, of which 15 were eligible for inclusion, including 8 cross-sectional studies, 5 case-control studies, and 2 cohort studies. The majority of studies enrolled both men and women, with mean age ranging from 41 to 66 years. In cross-sectional studies, the presence of apical periodontitis was significantly associated with cardiovascular disease with a combined odds ratio of 1.53 (95% CI: 1.02-2.29, p = 0.039; I2 = 75.0%; p < 0.001). In the case-control studies, the combined odds ratio did not show a significant association of apical periodontitis with cardiovascular disease (OR = 1.24; 95% CI: 0.67-2.29, p = 0.494; I2 = 82.1%; p < 0.001). The pooled risk ratio from the 2 cohort studies showed (RR = 1.27; 95% CI: 0.71-2.27, p = 0.413; I2 = 69.1%; p = 0.072) also showed no significant association between apical periodontitis and cardiovascular disease. These results should be interpreted with caution due to the high heterogeneity. Data derived from cross-sectional studies suggest a weak association between apical periodontitis and cardiovascular disease. As the results were not consistent across study designs, further research is recommended, namely longitudinal studies with long-term follow-up. PROSPERO database (CRD42020204379).


Brain Development in Fetuses of Mothers with Diabetes: A Case-Control MR Imaging Study.
Offspring exposed to maternal diabetes are at increased risk of neurocognitive impairment, but its origins are unknown. With MR imaging, we investigated the feasibility of comprehensive assessment of brain metabolism (1H-MRS), microstructure (DWI), and macrostructure (structural MRI) in third-trimester fetuses in women with diabetes and determined normal ranges for the MR imaging parameters measured. Women with singleton pregnancies with diabetes (n = 26) and healthy controls (n = 26) were recruited prospectively for MR imaging studies between 34 and 38 weeks' gestation. Data suitable for postprocessing were obtained from 79%, 71%, and 46% of women for 1H-MRS, DWI, and structural MRI, respectively. There was no difference in the NAA/Cho and NAA/Cr ratios (mean [SD]) in the fetal brain in women with diabetes compared with controls (1.74 [0.79] versus 1.79 [0.64], P = .81; and 0.78 [0.28] versus 0.94 [0.36], P = .12, respectively), but the Cho/Cr ratio was marginally lower (0.46 [0.11] versus 0.53 [0.10], P = .04). There was no difference in mean [SD] anterior white, posterior white, and deep gray matter ADC between patients and controls (1.16 [0.12] versus 1.16 [0.08], P = .96; 1.54 [0.16] versus 1.59 [0.20], P = .56; and 1.49 [0.23] versus 1.52 [0.23], P = .89, respectively) or volume of the cerebrum (243.0 mL [22.7 mL] versus 253.8 mL [31.6 mL], P = .38). Acquiring multimodal MR imaging of the fetal brain at 3T from pregnant women with diabetes is feasible. Further study of fetal brain metabolism in maternal diabetes is warranted.


Statins: immunomodulators for autoimmune rheumatic disease?
Inhibitors of 3-hydroxy-3-methylgluttaryl coenzyme A (HMG-CoA) reductase, or statins, are used extensively to reduced elevated lipid levels and reduce cardiovascular risk. However, accumulated evidence suggests that stains not only act by lowering cholesterol levels, but also exert pleiotropic effects on many essential cellular functions including cell proliferation, differentiation, and survival and participate in the regulation of cell shape and motility. Thus cardiovascular benefit is provided by lowering raised cholesterol levels and by modulation of the inflammatory component of this disease. Such an anti-inflammatory effect may also benefit patients with autoimmune rheumatic disease. This overview assesses the evidence for using statins in patients with rheumatoid arthritis and systemic lupus erythematosus (SLE).


Production of bioemulsifier by Acinetobacter species isolated from healthy human skin.
Six Acinetobacter sp. isolated from healthy human skin were checked for the production of bioemulsifier. Optimization studies indicated that Luria Bertani broth pH 7 supplemented with calcium chloride (1%) was the optimum medium. Temperature at 37 degrees C was optimum and inducer oils in the medium did not enhance bioemulsifier production. Partial purification of bioemulsifier and chemical analysis revealed that it is a proteoglycan with protein (53%), polysaccharide (43%) and lipid (2%). Maximum emulsification activity obtained was 400 EU/ml. Thin layer chromatography revealed the presence of mannose and rhamnose sugar and oleic and palmitic acids as parts of lipids. The yield obtained was 1.9 g / 1. Reconstitution studies revealed that the protein and polysaccharide fractions together display 94.55% of emulsification activity. It was also noted that the bioemulsifier was stable for 72 hr at 37 degrees C and displayed good cleaning property towards different oils. The partially purified bioemulsifier formed stable oil-in-water emulsions with plant oils.


Improving the safety of health information technology requires shared responsibility: It is time we all step up.
In 2011, an Institute of Medicine report on health information technology (IT) and patient safety highlighted that building health-IT for safer use is a shared responsibility between key stakeholders including: "vendors, care providers, healthcare organizations, health-IT departments, and public and private agencies". Use of electronic health records (EHRs) involves all these stakeholders, but they often have conflicting priorities and requirements. Since 2011, the concept of shared responsibility has gained little traction and EHR developers and users continue to attribute the substantial, long list of problems to each other. In this article, we discuss how these key stakeholders have complementary roles in improving EHR safety and must share responsibility to improve the current state of EHR use. We use real-world safety examples and outline a comprehensive shared responsibility approach to help guide development of future rules, regulations, and standards for EHR usability, interoperability and security as outlined in the 21st Century Cures Act. This approach clearly defines the responsibilities of each party and helps create appropriate measures for success. National and international policymakers must facilitate the local organizational and socio-political climate to stimulate the adoption of shared responsibility principles. When all major stakeholders are sharing responsibility, we will be more likely to usher in a new age of progress and innovation related to health IT.


Initial empirical antimicrobial therapy: duration and subsequent modifications.
Neutropenic patients at low risk of complications can receive oral ciprofloxacin or levofloxacin as outpatients. These agents plus amoxicillin/clavulanate or other penicillins, cephalosporins, or penem compounds are indicated to treat infections with gram-positive organisms in patients with oral mucositis or skin lesions. Parenteral fourth-generation cephalosporins or carbapenems can be given. For high-risk patients, monotherapy with cefepime or the carbapenems can be used. Methicillin-resistant Staphylococcus aureus should be treated with vancomycin or teicoplanin. For combination therapy, a third- or fourth-generation cephalosporin or carbapenem plus an aminoglycoside is desirable. Defervescence in 3-5 days for at least 7 days is suggested for subsequent management. Initial antibiotic(s) can be continued for patients who remain in good condition. For persistent fever after 3-5 days, the patient should be thoroughly reassessed. An aminoglycoside should be added for those initially treated with monotherapy. The initial cephalosporin can be changed to another cephalosporin or a carbapenem, or the initial carbapenem can be changed to a broad-spectrum cephalosporin. For patients initially receiving dual therapy, the cephalosporin or carbapenem can be changed as with monotherapy, whereas the initial aminoglycoside should be changed to another aminoglycoside or intravenous ciprofloxacin.


Bimonthly chemotherapy with oxaliplatin, irinotecan, infusional 5-fluorouracil/folinic acid in patients with metastatic colorectal cancer pretreated with irinotecan- or oxaliplatin-based chemotherapy.
This study was conducted to assess the tolerability and efficacy of a ternary bimonthly irinotecan (CPT-11) - oxaliplatin (OHP) - infusional 5-fluorouracil (5-FU)/folinic acid (FA) combination in advanced colorectal cancer patients who had received prior CPT-11 and/or OHP-based chemotherapy regimen. Colorectal cancer patients were given bimonthly CPT-11 as a 90-min infusion, followed by OHP (85 mg/m(2)), FA (200 mg/m(2)) 2-h infusions and 5-FU (48-h infusion). CPT-11 and 5-FU doses were escalated as reported below. 26 patients were recruited. Fourteen patients had received a prior CPT-11-, 6 patients a prior OHP-based chemotherapy regimen and 6 patients both regimens. Three dose levels were investigated: CPT-11 100, 120 and 140 mg/m(2) and 5-FU 1500, 1800 and 2100 mg/m(2) in 6, 12 and 8 patients, respectively. All patients were evaluable for toxicity, 24 for antitumor activity. At all dose levels toxicity was acceptable. Grade 4 toxicity occurred in two patients only (neutropenia in one case and stomatitis in another one, 3.8%). Grade 3 toxicities included nausea and vomiting (34.6%), asthenia (26.9%), neurosensory toxicity (15.4%), neutropenia (3.8%) and diarrhea (3.8%). Hematological toxicity was infrequent and generally mild. At the third dose level, a higher, although not significantly different incidence of hematological and neurosensory toxicity (both occurring in 62.5% of cases, all grades) was observed compared to the other two, while nausea and vomiting were significantly less frequent (37.5% vs 100%). Overall, we observed 2 complete responses, 9 partial responses (OR 45.8%), 8 stable disease (33.3%), and 5 disease progression (20.8%). Median overall survival was 18 months and median time-to-progression 5.5 months. This combination showed moderate toxicity and promising antitumor activity in CPT-11 and/or OHP pretreated colorectal cancer patients. The second dose level using CPT-11 at 120 mg/m(2) and 5-FU at 1800 mg/m(2) is recommended for further phase II studies in this patient population.


Functionalization of Endohedral Metallofullerenes with Reactive Silicon and Germanium Compounds.
Exohedral derivatization of endohedral metallofullerenes (EMFs) has been exploited as a useful method for characterizing the structural and chemical properties of EMFs, and for functionalizing them for potential applications. The introduction of heteroatoms, such as electropositive silicon atoms, to fullerene cages is a novel functionalization method that remarkably affects the electronic characteristics of fullerenes. This review comprehensively describes the results of the reactions of monometallofullerene, dimetallofullerene, and trimetallic nitride template EMFs with disilirane, silirane, silylene, and digermirane, which afforded the corresponding silylated and germylated fullerenes. Several examples emphasize that exohedral functionalization regulates the dynamic behaviors of the encapsulated metal atoms and clusters in the fullerene cages. The electronic effects of silyl and germyl groups are represented by comparing the redox properties of silylated and germylated EMFs with those of other EMFs derivatized with carbon-atom-based functional groups.


Cell separation: Terminology and practical considerations.
Cell separation is a powerful tool in biological research. Increasing usage, particularly within the tissue engineering and regenerative medicine communities, means that researchers from a diverse range of backgrounds are utilising cell separation technologies. This review aims to offer potential solutions to cell sorting problems and to clarify common ambiguities in terminology and experimental design. The frequently used cell separation terms of 'purity', 'recovery' and 'viability' are discussed, and attempts are made to reach a consensus view of their sometimes ambiguous meanings. The importance of appropriate experimental design is considered, with aspects such as marker expression, tissue isolation and original cell population analysis discussed. Finally, specific technical issues such as cell clustering, dead cell removal and non-specific antibody binding are considered and potential solutions offered. The solutions offered may provide a starting point to improve the quality of cell separations achieved by both the novice and experienced researcher alike.


Reversing thoracic hyperkyphosis: a case report featuring mirror image® thoracic extension rehabilitation.
[Purpose] To present a case of non-surgical reduction of thoracic hyperkyphosis utilizing a multimodal rehabilitation program emphasizing the mirror image® concept. [Subject and Methods] A 15-year-old female presented to a rehabilitation office suffering from back and neck pains and headaches. The patient was treated sporadically over a period of 13-months. Treatment consisted of anterior thoracic translation and thoracic extension exercises, spinal traction and spinal manipulation. [Results] After 13-months of treatment the patient displayed a significant reduction in hyperkyphosis and a dramatic correction of her overall posture and spine alignment corresponding to the reduction in back/neck pains, headaches and the simultaneous improvement of various other health issues. [Conclusion] Thoracic hyperkyphosis can be reduced through a multimodal rehabilitation program emphasizing mirror image thoracic extension procedures.


Complex differential variance angiography with noise-bias correction for optical coherence tomography of the retina.
Complex differential variance (CDV) provides phase-sensitive angiographic imaging for optical coherence tomography (OCT) with immunity to phase-instabilities of the imaging system and small-scale axial bulk motion. However, like all angiographic methods, measurement noise can result in erroneous indications of blood flow that confuse the interpretation of angiographic images. In this paper, a modified CDV algorithm that corrects for this noise-bias is presented. This is achieved by normalizing the CDV signal by analytically derived upper and lower limits. The noise-bias corrected CDV algorithm was implemented into an experimental 1 μm wavelength OCT system for retinal imaging that used an eye tracking scanner laser ophthalmoscope at 815 nm for compensation of lateral eye motions. The noise-bias correction improved the CDV imaging of the blood flow in tissue layers with a low signal-to-noise ratio and suppressed false indications of blood flow outside the tissue. In addition, the CDV signal normalization suppressed noise induced by galvanometer scanning errors and small-scale lateral motion. High quality cross-section and motion-corrected en face angiograms of the retina and choroid are presented.


Evolution of the pericardiocentesis technique.
: Pericardiocentesis is a valuable technique for the diagnosis and treatment of patients with pericardial effusion and cardiac tamponade, although it may be associated with potentially serious complications. Through the years, many different imaging approaches have been described to reduce the complication rate of the procedure. This systematic review provides a focused overview of the different techniques developed in recent years to reduce the procedural complications and to increase the related success rate.


Efficient and Accurate Identification of Platinum-Group Minerals by a Combination of Mineral Liberation and Electron Probe Microanalysis with a New Approach to the Offline Overlap Correction of Platinum-Group Element Concentrations.
Identification and accurate characterization of platinum-group minerals (PGMs) is usually a very cumbersome procedure due to their small grain size (typically below 10 µm) and inconspicuous appearance under reflected light. A novel strategy for finding PGMs and quantifying their composition was developed. It combines a mineral liberation analyzer (MLA), a point logging system, and electron probe microanalysis (EPMA). As a first step, the PGMs are identified using the MLA. Grains identified as PGMs are then marked and coordinates recorded and transferred to the EPMA. Case studies illustrate that the combination of MLA, point logging, and EPMA results in the identification of a significantly higher number of PGM grains than reflected light microscopy. Analysis of PGMs by EPMA requires considerable effort due to the often significant overlaps between the X-ray spectra of almost all platinum-group and associated elements. X-ray lines suitable for quantitative analysis need to be carefully selected. As peak overlaps cannot be avoided completely, an offline overlap correction based on weight proportions has been developed. Results obtained with the procedure proposed in this study attain acceptable totals and atomic proportions, indicating that the applied corrections are appropriate.


Mechanism and implications of CXCR4-mediated integrin activation by Porphyromonas gingivalis.
In monocytes and macrophages, the interaction of Porphyromonas gingivalis with Toll-like receptor 2 (TLR2) leads to the activation of a MyD88-dependent antimicrobial pathway and a phosphatidylinositol-3 kinase (PI3K) -dependent pro-adhesive pathway, which activates the β2 -integrin complement receptor 3 (CR3). By means of its fimbriae, P. gingivalis binds CXC-chemokine receptor 4 (CXCR4) and induces crosstalk with TLR2 that inhibits the MyD88-dependent antimicrobial pathway. In this paper, we investigated the impact of the P. gingivalis-CXCR4 interaction on the pro-adhesive pathway. Using human monocytes, mouse macrophages, or receptor-transfected cell lines, we showed that the binding of P. gingivalis fimbriae to CXCR4 induces CR3 activation via PI3K, albeit in a TLR2-independent manner. An isogenic strain of P. gingivalis expressing mutant fimbriae that do not interact with CXCR4 failed to efficiently activate CR3, leading to enhanced susceptibility to killing in vivo compared with the wild-type organism. This in vivo observation is consistent with previous findings that activated CR3 mediates safe entry of P. gingivalis into macrophages. Taken together with our previous work, these results indicate that the interaction of P. gingivalis with CXCR4 leads to inhibition of antimicrobial responses and enhancement of pro-adhesive responses, thereby maximizing its adaptive fitness in the mammalian host.


In vitro and in vivo activity of analogues of the kinin B2 receptor antagonist MEN1 1270.
In this study, we describe the in vitro and in vivo activities of a series of cyclic peptide analogues of the selective kinin B2 receptor antagonist MEN11270 on Chinese hamster ovary cells expressing the human B2 receptor (hB2R), the human isolated umbilical vein (hUV), the isolated guinea pig ileum (gpI), and bradykinin (BK) induced bronchoconstriction (BC) and hypotension in anaesthetized guinea pigs. Substitutions in the backbone of MEN1 1270 (H-DArg-Arg-Pro-Hyp-Gly-Thi-c(Dab-DTic-Oic-Arg)c(7gamma-10alpha)) aimed to increase the potency in inhibiting bronchospasm versus hypotension following the topical (intratracheal (i.t.)) or systemic (intravenous (i.v.)) application of these antagonists. A series of analogues were left unprotected from N-terminal cleavage by aminopeptidases (MEN12739, MEN13052, MEN13346, and MEN13371): these compounds maintained sizeable affinities for the hB2R (pKi = 9.4, 9.6, 9.7, and 8.6, respectively) and antagonist activities toward BK in the hUV (pA2 = 7.9, 8.3, 8.2, and 7.5) and gpI assays (pK(B) = 7.4, 7.8, 7.9, and 7.9), but the inhibition of BK-induced BC and hypotension in vivo was negligible following either i.v. or i.t. administration. Two analogues (MEN12388 and MEN13405) could be potential substrates of angiotensin-converting enzyme: these have good activity in the hB2R (pKi = 9.5 and 8.9, respectively), hUV (pA2 = 8.2 for MEN12388), and gpI assays (pK(B) = 8.4 and 8.0) but an in vivo activity 10- to 30-fold lower than the parent compound MEN1 1270 (pKi = 9.4, pA2 = 8.1, pKB = 8.3) when given by either the i.v. or the i.t. route. Other analogues were functionalized with a quaternary ammonium Lys derivative (MEN13031, MEN12374, and the previously mentioned MEN13052) or with an ethyl group on Arg (MEN13655 and the previously mentioned MEN13346 and MEN13405) in order to hinder or facilitate local absorption. MEN13346 and MEN13031 (pKi = 9.7and 9.5, pA2 = 8.2 and 7.9, pKB = 7.9 and 8.5, respectively) were 10- to 30-fold less active in vivo than MEN1 1270, without improving the discrimination between BK-induced BC and hypotension after either systemic or topical administration. It is concluded that the decreased in vivo activities of cyclic analogues of MEN11270 on BK-induced BC and hypotension following either their intratracheal or their intravenous routes of administration might be due in large part to metabolic degradation.


Topical therapy of alopecia areata with squaric acid dibutylester.
The efficacy of the topical allergen squaric acid dibutylester (SADBE) was investigated in the treatment of twenty-six patients with alopecia areata or alopecia totalis. The patients had their disease for a mean duration of 8.3 years (range, 3 months to 27 years). Sensitization was attempted with 2% SADBE in acetone. Five individuals could not be sensitized. The twenty-one sensitized patients were treated with topical applications of 0.001% to 1.0% SADBE in acetone adjusted to produce and maintain dermatitis for an average of 21 weeks. Eleven (52%) had excellent responses consisting of complete regrowth in six and cosmetically acceptable regrowth in five. The average treatment time for regrowth was 11 weeks (range, 5-20 weeks). Ten (48%) had no clinical response after an average of 19 weeks of therapy (range, 4-42 weeks). Topical SADBE is an effective therapy for some patients with long-standing alopecia areata.


Analysis of mRNA coding for blood-stage antigens of a rodent malarial parasite, Plasmodium yoelii: mRNA coding for a possible protective antigen purify as poly A-.
We have analyzed mRNA coding for blood-stage antigens of Plasmodium yoelii by using cellfree translation of poly A+ and poly A- RNA in conjunction with immunoprecipitations. Most of the antigens recognized by mouse hyperimmune serum to P. yoelii were coded by poly A+ mRNA ranging in size from 15S to 28S. However, certain P. yoelii antigens, notably those with m.w. greater than 150 kilodaltons (kd), were coded by mRNA that purified as being poly A-. Antigens recognized by a protective monoclonal antibody (McAb) were coded by such operationally poly A- RNA. Three polypeptides apparently coded by different poly A- RNA were immunoprecipitated by this McAb. With the use of another McAb, a poly A+ mRNA of about 19S was identified as coding for a polypeptide of 46 kd synthesized in cellfree translation reactions. The same McAb recognized a 34 kd polypeptide in metabolically labeled polypeptides of P. yoelii. This antigen appeared to be processed in vivo but not in vitro. The observation that some mRNA of P. yoelii purify as being poly A- has significant implications for the construction of cDNA libraries that employ poly A+ mRNA of malarial parasites: if it applies to other species of plasmodia, some potentially important operationally poly A- mRNA may not be represented in such libraries.


Histopathologic change of pulp tissue after surgical trauma in monkeys.
The present study was designed to investigate the effect of surgical trauma on pulp tissue. Two-walled osseous defects were surgically created at the mesial sides of the bilateral mandibular second bicuspids of four Taiwan monkeys. The cementum and partial thickness of dentin were removed with diamond bur. At one side of the mandible, the perforation was made at the midpoint of the root surface facing the created osseous defect to allow the pulp tissue to communicate the defects. The flaps were readapted. The monkeys were sacrificed 6 months postoperatively and prepared for the histologic evaluation. The results showed that in three of the four perforated sites, cementum-like structures were formed at the perforated areas. The pulp tissue kept vital. But replacement resorption was taking place in fourth studied site.


HPV16 E6/E7 upregulates HIF-2α and VEGF by inhibiting LKB1 in lung cancer cells.
Long-term persistent infection of HPV16 E6/E7 is frequently associated with lung cancers, especially in non-smokers and in Asians. However, molecular mechanisms of HPV16 E6/E7 induction of lung cancer are not fully understood. Using bi-directional genetic manipulation and four well-established lung cancer cell lines, we showed HPV16 E6/E7 downregulated expression of liver kinase B1 at both protein and messenger RNA levels; liver kinase B1 downregulated hypoxia-inducible factor 2α at protein level but not at messenger RNA level, and hypoxia-inducible factor 2α upregulated vascular endothelial growth factor at both protein and messenger RNA levels. This is the first study to show hypoxia-inducible factor 2α as a downstream effector of liver kinase B1 in lung cancer cells. Our results indicate that HPV16 E6/E7 indirectly upregulated the expression of vascular endothelial growth factor by inhibition of liver kinase B1 expression and upregulation of hypoxia-inducible factor 2α expression, thus propose a human papillomavirus-liver kinase B1-hypoxia-inducible factor 2α-vascular endothelial growth factor axis for the tumorigenesis of lung cancer. Our study also provides new evidence to support the critical role of liver kinase B1 in the pathogenesis of human papillomavirus-related lung cancer and suggests novel therapeutic targets.


Placenta-Derived Adherent Stromal Cell Therapy for Hematopoietic Disorders: A Case Study of PLX-R18.
The ephemeral placenta provides a noncontroversial source of young, healthy cells of both maternal and fetal origin from which cell therapy products can be manufactured. The 2 advantages of using live cells as therapeutic entities are: (a) in their environmental-responsive, multifactorial secretion profile and (b) in their activity as a "slow-release drug delivery system," releasing secretions over a long time frame. A major difficulty in translating cell therapy to the clinic involves challenges of large-scale, robust manufacturing while maintaining product characteristics, identity, and efficacy. To address these concerns early on, Pluristem developed the PLacental eXpanded (PLX) platform, the first good manufacturing practice-approved, 3-dimensional bioreactor-based cell growth platform, to enable culture of mesenchymal-like adherent stromal cells harvested from the postpartum placenta. One of the products produced by Pluristem on this platform is PLX-R18, a product mainly comprising placental fetal cells, which is proven in vivo to alleviate radiation-induced lethality and to enhance hematopoietic cell counts after bone marrow (BM) failure. The identified mechanism of action of PLX-R18 cells is one of the cell-derived systemic pro-hematopoietic secretions, which upregulate endogenous secretions and subsequently rescue BM and peripheral blood cellularity, thereby boosting survival. PLX-R18 is therefore currently under study to treat both the hematopoietic syndrome of acute radiation (under the US Food and Drug Administration [FDA]'s Animal Rule) and the incomplete engraftment after BM transplantation (in a phase I study). In the future, they could potentially address additional hematological indications, such as aplastic anemia, myelodysplastic syndrome, primary graft failure, and acute or chronic graft versus host disease.


Involvement of c-Abl, p53 and the MAP kinase JNK in the cell death program initiated in A2E-laden ARPE-19 cells by exposure to blue light.
The lipofuscin fluorophore A2E has been shown to mediate blue light-induced damage to retinal pigmented epithelial (RPE) cells. To understand the events that lead to RPE cell apoptosis under these conditions, we explored signaling pathways upstream of the cell death program. Human RPE cells (ARPE-19) that had accumulated A2E were exposed to blue light to induce apoptosis and the involvement of the transcription factors p53 and c-Abl and the mitogen activated protein kinases p38 and JNK were examined. We found that A2E/blue light caused upregulation and phosphorylation of c-Abl, and upregulation of p53. Pretreatment with the c-Abl inhibitor STI571 and transfection with siRNA specific to c-Abl and p53 prior to irradiation reduced A2E/blue light-induced cell death. Gene and protein expression of JNK and p38 was upregulated in response to A2E/blue light. Treatment with the JNK inhibitor SP600125 before irradiation resulted in increase in cell death whereas inhibition of p38 with SB203580 had no effect. This study indicates that c-Abl and p53 are important for execution of the cell death program initiated in A2E-laden RPE cells exposed to blue light, while JNK might play an anti-apoptotic role.


DNA-based differentiation of the Ecuadorian cocoa types CCN-51 and Arriba based on sequence differences in the chloroplast genome.
Two cocoa types, Arriba and CCN-51, are being cultivated in Ecuador. With regard to the unique aroma, Arriba is considered a fine cocoa type, while CCN-51 is a bulk cocoa because of its weaker aroma. Because it is being assumed that Arriba is mixed with CCN-51, there is an interest in the analytical differentiation of the two types. Two methods to identify CCN-51 adulterations in Arriba cocoa were developed on the basis of differences in the chloroplast DNA. On the one hand, a different repeat of the sequence TAAAG in the inverted repeat region results in a different length of amplicons for the two cocoa types, which can be detected by agarose gel electrophoresis, capillary gel electrophoresis, and denaturing high-performance liquid chromatography. On the other hand, single nucleotide polymorphisms (SNPs) between the CCN-51 and Arriba sequences represent restriction sites, which can be used for restriction fragment length polymorphism analysis. A semi-quantitative analysis based on these SNPs is feasible. A method for an exact quantitation based on these results is not realizable. These sequence variations were confirmed for a comprehensive cultivar collection of Arriba and CCN-51, for both bean and leaf samples.


Impact of adjustment measures on reducing outpatient waiting time in a community hospital: application of a computer simulation.
As an important determinant of patient satisfaction, waiting time, has gained increasing attention in the field of health care services. The present study aimed to illustrate the distribution characteristics of waiting time in a community hospital and explore the impact of potential measures to reduce outpatient waiting time based on a computer simulation approach. During a one-month study period in 2006, a cross-sectional study was conducted in a community hospital located in Shanghai, China. Baseline data of outpatient waiting time were calculated according to the records of registration time and payment time. A simulation technique was adopted to investigate the impact of perspective reform methods on reducing waiting time. Data from a total of 10,092 patients and 26,816 medical consultations were collected in the study and 19,947 medical consultations were included. The average of the total visit time for outpatients in this hospital was 43.6 minutes in the morning, 19.1 minutes in the afternoon, and 34.3 minutes for the whole day studied period. The simulation results suggested that waiting time for outpatients could be greatly reduced through the introduction of appointment system and flexible demand-orientated doctor scheduling according to the numbers of patients waiting at different time of the workday. Adoption of an appointment system and flexible management of doctor scheduling may be effective way to achieve decreased waiting time.


Cost of Intravenous Analgesia for the Management of Acute Pain in the Emergency Department is Substantial in the United States.
Background: Pain is a leading cause of admission to the emergency department (ED) and moderate-to-severe acute pain in medically supervised settings is often treated with intravenous (IV) opioids. With novel noninvasive analgesic products in development for this indication, it is important to assess the costs associated with IV administration of opioids. Materials and Methods: A retrospective observational study of data derived from the Premier database was conducted. All ED encounters of adult patients treated with IV opioids during a 2-year time period, who were charged for at least one IV opioid administration in the ED were included. Hospital reported costs were used to estimate the costs to administer IV opioids. Results: Over a 24 month-period, 7.3 million encounters, which included the administration of IV opioids took place in 614 US EDs. The mean cost per encounter of IV administration of an initial dose of the three most frequently prescribed opioids were: morphine $145, hydromorphone $146, and fentanyl $147. The main driver of the total costs is the cost of nursing time and equipment cost to set up and maintain an IV infusion ($140 ± 60). Adding a second dose of opioid, brings the average costs to $151-$154. If costs associated with the management of opioid-related adverse events and IV-related complications are also added, the total costs can amount to $269-$273. Of these 7.3 million encounters, 4.3 million (58%) did not lead to hospital admission of the patient and, therefore, the patient may have only required an IV catheter for opioid administration. Conclusions: IV opioid use in the ED is indicated for moderate-to-severe pain but is associated with significant costs. In subjects who are discharged from the ED and may not have required an IV for reasons other than opioid administration, rapid-onset analgesics for moderate-to-severe pain that do not require IV administration could lead to direct cost reductions and improved care.


Controlling fermentation of lignocellulose hydrolysates in a continuous hollow-fiber reactor using biosensors.
Fermentation of lignocellulose hydrolysates as spent sulfite liquor or as hydrolysate from sulfur-dioxide-treated wood to ethanol has been controlled by using biosensors for glucose and ethanol. Yield and productivity were studied with respect to concentration level of the metabolites in a continuous hollow-fiber reactor. High constant yield was achieved by controlling the glucose to low concentration levels. Reduced productivity were obtained when fermenting at high ethanol concentrations as an effect of inhibition of the yeast cells. The observations emphasize the advantage of controlling the process to favorable concentrations of monosaccharides and ethanol.


[Clinical treatment outcome of grade III, non-small cell bronchogenic carcinoma using "whole-body" gamma-knife system].
The results of treatment of 80 patients with non-small cell lung cancer using the Chinese-made "Whole-Body" gamma-knife system were analyzed. Primary focus and involved lymph nodes were exposed. Neither gap between primary focus and involved lymph node or nodes, nor organs of the mediastinum were exposed. Exposure regimens were: STD = 4 Gy (2.5 - 10 Gy) 5 times a week; TTD=48 Gy (27-52 Gy) per primary focus and 4 Gy (3-6 Gy) 5 times a week; TTD = 43.5 Gy (30-52 Gy) per lymph nodes (105.6 - 150 Gy = equ.). Complete response was observed in 27.5% (22/80), partial - 42.5% (34/80). Overall survival was 61.3% among those sick for one year; 41.5%--2 years and 20.75%--3 years (mean survival time--20 months).


The disturbance of hemostasis in septic shock: role of neutrophil elastase and thrombin, effects of antithrombin III and plasma substitution.
In 42 patients with septic shock, 29 of whom underwent substitution with antithrombin III concentrate and fresh frozen plasma for coagulation disorders, the proteinase-inhibitor complexes thrombin-antithrombin III and neutrophil elastase-alpha 1 proteinase inhibitor, were elevated on admission. On admission, the elastase complex was significantly higher in the patients receiving substitution (p = 0.0039), but at the endpoint it was higher in the non-survivors (p = 0.0040). The elastase decrease was confined to the substitution group with the thrombin complex decreasing in both groups. Initially the thrombin complex correlated with prothrombin times and factor XIII, while the elastase complex correlated with creatinine, thrombocyte count and prothrombin times in the late stages. Hemostatic disturbance, thrombin generation and neutrophil elastase release were favorably influenced by substitution. Furthermore, in this uncontrolled pilot study, the survival rate was higher in the treated (16 of 29) than in the untreated (1 of 13) patients, although the treated patients initially had pronounced hemostatic disturbances.


Xanthogranulomatous Pyelonephritis of a Lower Pole Moiety in a Duplicated Collecting System Kidney.
Xanthogranulomatous pyelonephritis (XGP) is a destructive inflammatory process which is frequently caused by recurrent urinary tract infections or chronic obstruction by kidney stones. We present a 56-year-old female with an extensive retroperitoneal urinoma and xanthogranulomatous pyelonephritis of the lower pole moiety in a kidney with a duplicated collecting system due to obstructive nephrolithiasis. After drainage of the urinoma, the patient underwent a definitive lower pole heminephrectomy with preservation of the functional upper pole. We review important clinical features of xanthogranulomatous pyelonephritis and considerations for surgery on a duplicated kidney.


A Practical Suggestion for Prepare Dorsal Onlay Graft.
Nowadays, rhinoplasty is one of the most popular surgical procedures. Dorsal contour irregularities caused by various maneuvers, such as hump resection, are a major concern in patients who have undergone rhinoplasty. The most common graft used in this case is dorsal onlay graft which is made from sliced and crushed cartilage. Ear, nose and throat specialists usually use Swester table (mayo desk) for preparing the graft, if there is no other steril metal instrument. Crushed cartilage is done on a sterile gauze or on the tables' cover, as a result cartilage may be contaminated with particules from the tables' cover and sterile gauze.The authors recommend using the steril pack of a new sterilized surgical instrument opened on the table for the slicing or crushing process. In this way, the cartilage can be spared from contamination and the loss of some cartilage to the table during slicing or crushing can be prevented.


Chief executives. Off limits.
Trust remuneration committees are paying chief executives above the limits recommended in Department of Health guidance. In doing so they are ignoring the government's stated policy of fair pay for all in the NHS and their duty of accountability. Excessive awards made by a remuneration committee can be subject to judicial review and overturned. The health secretary should review the workings of trust remuneration committees and ensure that their decisions are transparent to the public.


3-Methyl glutaconic aciduria in Iraqi Jewish children may be misdiagnosed as cerebral palsy.
It is generally accepted that patients with cerebral palsy suffer from a static encephalopathy causing a non-progressive disorder of posture and/or movement. We describe 7 patients from 5 families who were initially diagnosed with cerebral palsy. Eventually, excessive excretion of urinary 3-methyl glutaconic acid (3-MGA) was found. The data of our 7 patients are quite similar to the clinical description of Costeff et al (1989): Jewish-Iraqi origin (7/7), consanguinity (2/7), involuntary movements (5/7), ataxia (6/7), pyramidal involvement (6/7) and optic atrophy (6/7). The cognitive functions were intact in 5/7 and 2/7 showed mild to moderate mental retardation. The mean delay in the definitive diagnosis was 9 years. Cerebral palsy-like symptoms accompanied by optic atrophy and extrapyramidal signs should call for extensive metabolic evaluation including the determination of urinary 3-MGA.


Prothrombin conversion under flow conditions by prothrombinase assembled on adherent platelets.
Prothrombin activation by prothrombinase was investigated on platelets adhered onto a fibrinogen-coated coverslip mounted in a flow chamber. Once bound to the fibrinogen, platelets gradually changed their morphology: they developed pseudopods, spread over the surface and finally transformed into balloon-shaped cells. This last morphologic change required the presence of calcium and was accompanied by the exposure of procoagulant phospholipid at the outer membrane as detected by the capability of the platelets to bind fluorescein-labelled annexin V. Prothrombinase complexes were allowed to assemble on these adhered platelets by perfusion with factor Xa and varying concentrations of factor Va and prothrombin. The steady-state rate of thrombin formation during continuous flow increased with the prothrombin concentration but not with the factor Va between 0.05 and 0.5 nM. Once prothrombinase was assembled, factor Xa could be omitted from the perfusion mixture without affecting the steady state rate of thrombin production. Our study demonstrates the efficient ability of the procoagulant surface of adherent platelet to support the assembly of stable prothrombinase complexes. Thrombin production was limited by the rate of supply of prothrombin towards the catalytic surface.


Computational fluid dynamics in the evaluation of hemodynamic performance of cavopulmonary connections after the Norwood procedure for hypoplastic left heart syndrome.
Computational fluid dynamics have been used to study the hemodynamic performance of surgical operations, resulting in improved design. Efficient designs with minimal energy losses are especially important for cavopulmonary connections. The purpose of this study was to compare hydraulic performance between the hemi-Fontan and bidirectional Glenn procedures, as well as the various types of completion Fontan operations. Three-dimensional models were constructed of typical hemi-Fontan and bidirectional Glenn operations according to anatomic data derived from magnetic resonance scans, angiocardiograms, and echocardiograms. Boundary conditions were imposed, and fluid dynamics were calculated from a mathematic code. Power losses, flow distribution to each lung, and pressures were measured at three predetermined levels of pulmonary arteriolar resistance. Models of the lateral tunnel, total cavopulmonary connection, and extracardiac conduit completion Fontan operations were constructed, and power losses, total flow distribution, vena caval and pulmonary arterial pressures, and flow distribution of inferior vena caval return were calculated. The hemi-Fontan and bidirectional Glenn procedures performed nearly identically, with similar power losses and nearly equal flow distributions to each lung at all levels of pulmonary arteriolar resistance. However, the lateral tunnel Fontan procedure as performed after the hemi-Fontan operation had lower power losses (6.9 mW, pulmonary arteriolar resistance 3 units) than the total cavopulmonary connection (40.5 mW) or the extracardiac conduit (42.9 mW), although the inclusion of an enlargement patch toward the right in the total cavopulmonary connection was effective in reducing the difference (10.0 mW). Inferior vena caval flow to the right lung was 52% for the lateral tunnel, compared with 19%, 30%, 19%, and 15% for the total cavopulmonary connection, total cavopulmonary connection with right-sided enlargement patch, extracardiac conduit, and extracardiac conduit with a bevel to the left lung, respectively. According to these methods, the hemi-Fontan and bidirectional Glenn procedures performed equally well, but important differences in energy losses and flow distribution were found after the completion Fontan procedures. The superior hydraulic performance of the lateral tunnel Fontan operation after the hemi-Fontan procedure relative to any other method may be due to closer to optimal caval offset achieved in the surgical reconstruction.


Unfolding dynamics of the mucin SEA domain probed by force spectroscopy suggest that it acts as a cell-protective device.
MUC1 and other membrane-associated mucins harbor long, up to 1 μm, extended highly glycosylated mucin domains and sea urchin sperm protein, enterokinase and agrin (SEA) domains situated on their extracellular parts. These mucins line luminal tracts and organs, and are anchored to the apical cell membrane by a transmembrane domain. The SEA domain is highly conserved and undergoes a molecular strain-dependent autocatalytic cleavage during folding in the endoplasmic reticulum, a process required for apical plasma membrane expression. To date, no specific function has been designated for the SEA domain. Here, we constructed a recombinant protein consisting of three SEA domains in tandem and used force spectroscopy to assess the dissociation force required to unfold individual, folded SEA domains. Force-distance curves revealed three peaks, each representing unfolding of a single SEA domain. Fitting the observed unfolding events to a worm-like chain model yielded an average contour length of 32 nm per SEA domain. Analysis of forces applied on the recombinant protein revealed an average unfolding force of 168 pN for each SEA domain at a loading rate of 25 nN·s(-1). Thus, the SEA domain may act as a breaking point that can dissociate before the plasma membrane is breached when mechanical forces are applied to cell surfaces.


Requirement for annexin A1 in plasma membrane repair.
Ca2+ entering a cell through a torn or disrupted plasma membrane rapidly triggers a combination of homotypic and exocytotic membrane fusion events. These events serve to erect a reparative membrane patch and then anneal it to the defect site. Annexin A1 is a cytosolic protein that, when activated by micromolar Ca2+, binds to membrane phospholipids, promoting membrane aggregation and fusion. We demonstrate here that an annexin A1 function-blocking antibody, a small peptide competitor, and a dominant-negative annexin A1 mutant protein incapable of Ca2+ binding all inhibit resealing. Moreover, we show that, coincident with a resealing event, annexin A1 becomes concentrated at disruption sites. We propose that Ca2+ entering through a disruption locally induces annexin A1 binding to membranes, initiating emergency fusion events whenever and wherever required.


Universality and Stability Phase Diagram of Two-Dimensional Brittle Fracture.
The two-dimensional oscillatory crack instability, experimentally observed in a class of brittle materials under strongly dynamic conditions, has been recently reproduced by a nonlinear phase-field fracture theory. Here, we highlight the universal character of this instability by showing that it is present in materials exhibiting widely different near crack tip elastic nonlinearity, and by demonstrating that the oscillations wavelength follows a universal master curve in terms of dissipation-related and nonlinear elastic intrinsic length scales. Moreover, we show that upon increasing the driving force for fracture, a high-velocity tip-splitting instability emerges, as experimentally demonstrated. The analysis culminates in a comprehensive stability phase diagram of two-dimensional brittle fracture, whose salient properties and topology are independent of the form of near tip nonlinearity.


Oligosaccharide sequences in Quillaja saponins by electrospray ionization ion trap multiple-stage mass spectrometry.
Ten different samples with 13 previously identified saponin structures from Quillaja saponaria Molina were investigated by electrospray ionization ion trap multiple-stage mass spectrometry (ESI-ITMS(n)) in positive and negative ion modes. Both positive and negative ion mode MS(1)-MS(4) spectra were analyzed, showing that structural information on the two oligosaccharide parts in the saponin can be obtained from positive ion mode spectra whereas negative ion mode spectra mainly gave information on one of the oligosaccharide parts. Analysis of MS(1)-MS(4) spectra identified useful key fragment ions important for the structural elucidation of Quillaja saponins. A flowchart involving a stepwise procedure based on key fragments from MS(1)-MS(3) spectra was constructed for the identification of structural elements in the saponin. Peak intensity ratios in MS(3) spectra were found to be correlated with structural features of the investigated saponins and are therefore of value for the identification of terminal monosaccharide residues.


[Detection and clinical significance of PML protein expression of acute promyelocytic leukemia cells].
Objective To study the expression patterns of promyelocytic leukemia (PML) protein at different stages of acute promyelocytic leukemia (APL) and investigate the effects that the arsenical, all-trans retinoic acid (ATRA) and chemotherapeutic drugs on PML protein expression patterns. Methods The bone marrow cells of 42 APL patients, NB4 cells and MR2 cells were collected and treated separately with 1 μmol/L arsenic trioxide (As2O3), 1 μmol/L tetra-arsenic tetra-sulfide (As4S4), 1 μmol/L ATRA, 10 mg/L cytosine arabinoside, and 1 mg/L homoharringtonine. The changes of fluorescent signals before and after the treatment were detected by indirect immunofluorescence technique. Results The 87.5% of initial onset APL cases showed small fluorescent signals (fluorescent signals in the nucleus look like a starry sky, more than 30 fluorescent light dots per cell); 91.67% of complete remission APL cases manifested large fluorescent signals (fluorescent signals in the nucleus are larger, fewer than 30 fluorescent light dots per cell); and 83.33% of relapse APL cases presented small fluorescent signals. Compared with control group without medication, the PML protein expression of APL cells in initial onset cases had larger fluorescent signals in the ATRA group, As2O3 group and As4S4 group, whereas there were no significant changes in the arabinoside cytosine group and the homoharringtonine group. In addition, ATRA transformed the PML protein expression of NB4 cells from small fluorescent signals into large ones, but did not change small fluorescent signals of the PML protein expression in MR2 cells. Also, As2O3 and As4S4 turned the PML protein expression of NB4 and MR2 cells from small fluorescent signals into large ones. Conclusion Detection of PML protein expression patterns of APL cells has practical application values for evaluating the prognosis of APL and making a rational choice of drugs.


Femoral artery pseudoaneurysm due to a gunshot injury.
Femoral artery pseudoaneurysms are commonly iatrogenic due to increasing use of the artery for arterial interventions. Other reasons of pseudoaneurysm formation are intravenous drug use and penetrating trauma. Here, we have discussed the management strategy of a femoral artery pseuodoaneursym and the modalities for preventing the misdiagnoses of the pseudoaneurysm in the emergency department. A 50-year-old male patient was referred to our emergency department (ED) with claudication and severe local swelling. Ten days earlier, he had been referred to another ED immediately after a gunshot injury to the left inguinal zone. Duplex ultrasound and CT angiography of the left lower extremity revealed a 4 cm sac of pseudoaneurysm on the distal part of posterofemoral branch of deep femoral artery and a 9*10 cm haematoma on the posteromedial part of pseudoaneurysm. The patient underwent open repair surgery due to co-existing large haematoma and risk of infection. The patient was discharged after three days hospitalization. Pain, extremity oedema, pulsatile mass, femoral bruit, palpable thrill, and compressive neuropathy should alert the physician to possible femoral artery pseudoaneurysm. Duplex ultrasound and CT angiography are important diagnostic steps to reveal a possible life-threatening vascular injury.


Effect of a Y265F mutant on the transamination-based cycloserine inactivation of alanine racemase.
The requirement for d-alanine in the peptidoglycan layer of bacterial cell walls is fulfilled in part by alanine racemase (EC 184.108.40.206), a pyridoxal 5'-phosphate (PLP)-assisted enzyme. The enzyme utilizes two antiparallel bases focused at the C(alpha) position and oriented perpendicular to the PLP ring to facilitate the equilibration of alanine enantiomers. Understanding how this two-base system is utilized and controlled to yield reaction specificity is therefore a potential means for designing antibiotics. Cycloserine is a known alanine racemase suicide substrate, although its mechanism of inactivation is based on transaminase chemistry. Here we characterize the effects of a Y265F mutant (Tyr265 acts as the catalytic base in the l-isomer case) of Bacillus stearothermophilus alanine racemase on cycloserine inactivation. The Y265F mutant reduces racemization activity 1600-fold [Watanabe, A., Yoshimura, T., Mikami, B., and Esaki, N. (1999) J. Biochem. 126, 781-786] and only leads to formation of the isoxazole end product (the result of the transaminase pathway) in the case of d-cycloserine, in contrast to results obtained using the wild-type enzyme. l-Cycloserine, on the other hand, utilizes a number of alternative pathways in the absence of Y265, emphasizing the importance of Y265 in both the inactivation and racemization pathway. In combination with the kinetics of inactivation, these results suggest roles for each of the two catalytic bases in racemization and inactivation, as well as the importance of Y265 in "steering" the chemistry to favor one pathway over another.


Prostate cancer stem cells.
Despite the discovery over 60 years ago by Huggins and Hodges that prostate cancers respond to androgen deprivation therapy, hormone-refractory prostate cancer remains a major clinical challenge. There is now mounting evidence that solid tumours originate from undifferentiated stem cell-like cells coexisting within a heterogeneous tumour mass that drive tumour formation, maintain tumour homeostasis and initiate metastases. This review focuses upon current evidence for prostate cancer stem cells, addressing the identification and properties of both normal and transformed prostate stem cells.


Mitochondrial abnormalities and low grade inflammation are present in the skeletal muscle of a minority of patients with amyotrophic lateral sclerosis; an observational myopathology study.
Amyotrophic lateral sclerosis (ALS) is a primary progressive neurodegenerative disease characterised by neuronal loss of lower motor neurons (in the spinal cord and brainstem) and/or upper motor neurons (in the motor cortex) and subsequent denervation atrophy of skeletal muscle. A comprehensive examination of muscle pathology from a cohort of clinically confirmed ALS patients, including an investigation of inflammation, complement activation, and deposition of abnormal proteins in order to compare them with findings from an age-matched, control group. 31 muscle biopsies from clinically confirmed ALS patients and 20 normal controls underwent a comprehensive protocol of histochemical and immunohistochemical stains, including HLA-ABC, C5b-9, p62, and TDP-43. Neurogenic changes were confirmed in 30/31 ALS cases. In one case, no neurogenic changes could be detected. Muscle fibre necrosis was seen in 5/31 cases and chronic mononuclear inflammatory cell infiltration in 5/31 (2 of them overlapped with those showing muscle necrosis). In four biopsies there was an increase in the proportion of cytochrome oxidase (COX) negative fibres (2-3%). p62 faintly stained cytoplasmic bodies in eight cases and none were immunoreactive to TDP-43. This large series of muscle biopsies from patients with ALS demonstrates neurogenic atrophy is a nearly uniform finding and that mild mitochondrial abnormalities and low-grade inflammation can be seen and do not rule out the diagnosis of ALS. These findings could lend support to the notion that ALS is a complex and heterogeneous disorder.


Lower bound of assortativity coefficient in scale-free networks.
The degree-degree correlation is important in understanding the structural organization of a network and dynamics upon a network. Such correlation is usually measured by the assortativity coefficient r, with natural bounds r∈[-1,1]. For scale-free networks with power-law degree distribution p(k)∼k-γ, we analytically obtain the lower bound of assortativity coefficient in the limit of large network size, which is not -1 but dependent on the power-law exponent γ. This work challenges the validation of the assortativity coefficient in heterogeneous networks, suggesting that one cannot judge whether a network is positively or negatively correlated just by looking at its assortativity coefficient alone.


[Current problems in psychiatric health care in Poland (2004)].
The paper is a presentation of the most important and actual psychiatric health care problems in Poland: the financial situation of hospitals, the risk faced by the out-patient psychiatric care and the ethical problems related to investigations of psychotropic drugs.


[A pilot study on the accuracy of digital model reconstructed by cone beam CT].
To study the feasibility of digital model produced by cone beam CT. Ten patients seeking orthodontic treatment with congested lower incisors were enrolled in this study. Plaster models of the lower arch were made using routing method. Digital model was reconstructed from these plaster models by NewTom 3G cone beam CT with 6 inch model. Linear measurements including right first premolar width, arch width, arch length, right first incisor width, all incisors and canines width, were done on the lower arch on plaster models with calipers. The same measurements on digital model were made using software Simplant Pro11.04. Linear measurements were repeated one week later on both plaster and digital model. Paired t test was used to determine the difference between the linear measurements on both models. The absolute measurements errors of two models were compared using paired t test. All statistical analysis was performed using SPSS13.0 software package. The linear measurements of the digital model slightly underestimated the real plaster model, but no significant difference of the linear measurements was seen between the plaster model and the digital model. The value of all of the lower incisors and canines width of plaster models was significantly higher than that of digital model. Mean absolute difference of different measurement of digital model was not significant from that of plaster model. Digital model using CBCT is feasible for the linear measurement and storage.


Imaging appearance of solitary fibrous tumor of the abdominopelvic cavity.
The computed tomographic or magnetic resonance imaging appearance of solitary fibrous tumors of the abdominopelvic cavity has previously only been presented in the English literature as individual case reports. In this article, we present the cross-sectional imaging appearance of 5 such cases, all of which exhibited highly similar imaging features, including well-circumscribed margins, lack of invasion of adjacent structures, and avid enhancement. In view of these shared imaging features, it may be possible to suggest the diagnosis preoperatively. Given their unpredictable biologic behavior with infrequent reports of recurrent or metastatic disease, complete surgical excision and long-term follow-up for these lesions is recommended.


Exercise exerts an anxiolytic effect against repeated restraint stress through 5-HT2A-mediated suppression of the adenosine A2A receptor in the basolateral amygdala.
Repeated or chronic stressful stimuli induce emotion- and mood-related abnormalities, such as anxiety and depression. Conversely, regular exercise exerts protective effects. Here, we found that exercise recovered anxiety-like behaviors, as measured using the open field and elevated plus maze tests in an anxiety mouse model. In addition to behavioral improvement, exercise enhanced the synaptic density of the 5-hydroxytryptamine 2A receptor (5-HT2AR), but not the 5-HT1AR in the basolateral amygdala (BLA) region in this mouse model. Furthermore, global treatment with a selective 5-HT2AR antagonist (MDL11930) generated an anxiety phenotype. Thus, synaptic recruitment of 5-HT2AR in BLA neurons may mediate the anxiolytic effects of exercise. The exercise regimen also reduced adenosine A2A receptor (A2AR)-mediated protein kinase A (PKA) activation, and the anxiolytic effect of the exercise was blunted by local activation of A2AR within the BLA using CGS21680, a selective A2AR agonist. Particularly, A2AR-mediated PKA activity was shown to be dependent on 5-HT2AR signaling in the BLA. These results imply that repeated stress upregulates A2AR-mediated adenosine signaling to facilitate PKA activation, whereas regular exercise inhibits A2AR function by increasing 5-HT2AR in the BLA. Accordingly, this integrated modulation of 5-HT and adenosine signaling, via 5-HT2AR and A2AR respectively, may be a mechanism underlying the anxiolytic effect of regular exercise.


Carcinogenicity of benzene hexachloride and its isomers.
Technical BHC and the alpha, beta, gamma and delta isomers of BHC are carcinogenic for the liver of mice and rats. Mice given the isomers of BHC developed carcinomas and hyperplastic nodules of the liver as early as 24 weeks. Several strains of mice (dd, ICR-JLC, CL1, DDY, IRC, DBA/Z, C3H/HEN, C57BL/6 were susceptible. Male mice were more susceptible to hepatic carcinogenesis than female mice and they appeared to be more susceptible to alpha-BHC. The incidence of neoplasms of the liver was increased when beta-, gamma-, or delta-BHC were each given together with alpha-BHC. PCB-5 promoted the induction of hepatic neoplasms when administered with beta-BHC. Technical BHC and its isomers are carcinogenic for the liver of male and female Wistar rats. Beta-BHC is carcinogenic for the liver of Osborne-Mendel male rats. Osborne-Mendel rats receiving delta-BHC developed cirrhosis of the liver, portal vein thrombosis, and focal necrosis of the skeletal muscle. Male rats ingesting technical BHC or the beta or delta isomers also had atrophic testes.


The effect of volume replacement during therapeutic leukapheresis on white blood cell reduction in patients with extreme leukocytosis.
Extreme leukocytosis, generally defined as a white blood cell (WBC) count of more than 100 × 10(9) /L consisting largely of blast cells, especially when accompanied by clinical signs and symptoms of leukostasis or hyperviscosity, often predicts a poor clinical outcome in patients with acute leukemia. In this study, we aimed to investigate the effect of volume replacement (VR) during therapeutic leukapheresis (TA) procedure on early mortality rate and WBC reduction. We retrospectively analyzed 29 patients who underwent TA from 2007 to 2011. Fifteen of the patients underwent TA procedure with VR and 14 of the patients underwent TA procedure without VR. WBC reduction was significantly higher in patients who underwent TA with VR (p < 0.001). Early mortality rate was significantly lower in leukemia patients who underwent TA with VR than in patients who underwent TA without VR (p < 0.01); early mortality rates were 6.7% for 7-day and 13.8% for 100-day survivals. The mortality rates in the TA without VR group, however, were 42.9 and 71.4% for 7- and 100-day survivals, respectively. Decreased early mortality rate in TA with VR group may be associated with prompt reduction of WBCs achieved with TA with VR and may also be associated with removal of the cytokines related to leukostasis. TA with VR would give more time for induction chemotherapy and increased overall survival rate.