Platelet-rich plasma loaded hydrogel scaffold enhances chondrogenic differentiation and maturation with up-regulation of CB1 and CB2.
Three-dimensional scaffolds like hydrogels can be used for cell and drug delivery and have become a major research focus in tissue engineering. Presently, we investigated the regenerative potency of platelet-rich plasma (PRP) combined with a chondrocyte/hydrogel composite scaffold in the repair of articular cartilage defects using a rabbit model. Primary isolated joint chondrocytes from the trachlear groove of rabbit were cultured in hydrogels as follows; hydrogel (2900 Pa or 5900 Pa)+chondrocytes and hydrogel+chondrocytes+PRP for in vitro analysis and in vivo implantation. The 5900 Pa hydrogel markedly increased cellular viability and development in a time-dependent manner. Furthermore, the hydrogels attenuated the expression of SOX-9, aggrecan, and type II collagen. PRP-containing hydrogels produced an immediate increase in mRNA levels of cannabinoid receptor (CB)1 and CB2, compared with control and PRP-free hydrogels. Osteochondral defects were enhanced recovery with formation of cartilage and perichondrium in the 5900 Pa hydrogel+chondrocytes+PRP. Hydrogel may provide a suitable environment for proliferation and maturation of joint chondrocytes in relation to the gelation density and bioactive sources like PRP resulting in improvement for cartilage regeneration.


Non-invasive measurement of biomechanical skin properties in systemic sclerosis.
To evaluate biomechanical properties of skin in patients with systemic sclerosis (SSc) using the BTC-2000 suction device. Twenty five patients with limited cutaneous SSc (lcSSc), 20 patients with diffuse disease (dcSSc), and 25 age matched healthy controls were evaluated. Viscoelastic deformation (VED, mm), elastic deformation (ED, mm), ultimate deformation (UD, mm), and pressure-deformation ratio (PDR, mm Hg/mm) were measured on the dorsal surface of the forearm, shoulder, and above the trapezius muscle on the back. Indices of skin extensibility (VED, ED, UD) were reduced and resistance to stress (PDR) increased in patients with dcSSc compared with healthy controls, or patients with lcSSc, at all three sites (p<0.001). At all sites, and overall, UD, ED, and VED were lower and PDR was higher at skin score above grade 2, compared with clinically normal skin. For both lcSSc and dcSSc biomechanical differences from controls were found even at sites of clinically normal skin. BTC-2000 is a sensitive tool for clinical evaluation of skin involvement in SSc and may be a valuable adjunct to skin sclerosis score in disease monitoring.


Structure of the Adenohypophysis in parturient female and neonate harp seals, Pagophilus groenlandicus.
The pars distalis of parturient harp seals (Pagophilus groenlandicus) is divisible into three distinct zones based on the major cellular components of the different regions. A clear zonation is lacking in neonate seals, the medical "basophilic wedge" and the anterior gonadotrophic were small and difficult to identify. Five cell types were identified based on cytological characteristics: somatotrophs, mammotrophs, thyrotrophs, gonadotrophs and stellate cells; corticotrophs could not be identified, with any certainty, in any of the preparations, nor could the gonadotrophs be classified into LH, FSH, and ICSH cells. The pars intermedia enlarged during the early post-partum period and was large in the parturient females.


Exosomes Derived from Breast Cancer Cells, Small Trojan Horses?
Exosomes are small extracellular vesicles secreted to the extracellular environment by several cell types, including tumor cells. It has been demonstrated that exosomes have an important role in intercellular communication, but they have recently been implicated in various tumor processes, including the oncogenic transformation of cells in the tumor microenvironment, tumor drug resistance, and the transport of tumor factors. Tumors appear to use exosomes to dialogue with and transform neighboring cells to create an ideal environment for their growth and expansion. On the other hand, the structure and function of exosomes may make them useful in cancer diagnosis and prognosis, because they contain molecules that could serve as biomarkers, including oncogenes, miRNAs, and certain proteins. They have the ability to travel via body fluids, from which they could be isolated and used to transport drugs to specific targets. This review aims to provide an update on the role of exosomes derived from breast cancer cells.


Role of single disulfide in recombinant human tumor necrosis factor-alpha.
Two analogs of tumor necrosis factor-alpha (TNF-alpha) were produced by in vitro site-directed mutagenesis. In these analogs, cysteine residues at positions 69 and 101, which form a disulfide bond, were changed to alanine or leucine. CD spectra showed that the analogs are apparently similar in secondary and tertiary structure to the natural sequence TNF-alpha. In addition, the molecular size of the analogs was identical to that of the natural sequence TNF-alpha as determined by gel filtration. However, fluorescence spectra and quenching indicated that the removal of the disulfide bond alters the local conformation around tryptophan residues. The cytolytic, macrophage activation, and lipogenic activities decreased in the order of the natural sequence TNF-alpha greater than the alanine analog greater than the leucine analog, suggesting that the surface involving the disulfide bond plays a role in these biological functions and the introduced modifications decrease the activity. Differential effect of the modifications was suggested in the antiviral activity, since in this assay only the leucine analog showed significantly lower activity.


IL12B rs6887695 polymorphism and interaction with alcohol intake in the risk of ulcerative colitis in Japan.
The interleukin (IL)-23/Th17 pathway plays a critical role in ulcerative colitis (UC). The IL-12p40 subunit, which is shared by IL-23 and IL-12, is encoded by the IL12B gene. The current case-control study investigated the association between IL12B SNP rs6887695 and the UC risk. There were 384 cases within 4 years of UC diagnosis and 661 controls who were enrolled. Adjustments were made for sex, age, pack-years of smoking, alcohol consumption, history of appendicitis, family history of UC, education level, and body mass index. Subjects with the GG IL12B SNP rs6887695 genotype had a significantly increased risk of UC compared with those with the CC genotype (adjusted odds ratio [AOR], 1.60; 95% confidence interval [CI], 1.08-2.36). This positive association was also significant using the additive and recessive models (AOR, 1.25; 95% CI, 1.03-1.52; AOR, 1.50; 95% CI, 1.08-2.09, respectively). An independent inverse relationship was observed between ever alcohol consumption and the UC risk in those with the CC genotype while no significant association was found in those with at least one G allele (P for interaction = 0.0008). IL12B SNP rs6887695 was significantly associated with UC. The influence of alcohol consumption might rely on rs6887695.


Origin of efferent fibers of the vestibular apparatus in goldfish. A horseradish peroxidase study.
After injecting a horseradish peroxidase (HRP) solution into the vestibular apparatus in goldfish (Carassius auratus), cells in the medulla oblongata exhibited retrogradely transported HRP reaction product. These labeled cells were confined to 3 nuclei, i.e. the nucleus motorius tegmenti of Bartelmez (with the most prominent labeled cell groups), the ventral vestibular nucleus (Dieter's nucleus) and the medial vestibular nucleus. We consider these labeled cells to be the origin of the efferent innervation to the vestibular apparatus in goldfish. Neurons providing efferent innervation to the sensory periphery in the nucleus motorius tegmenti have, as yet, not been described.


Somatotopic organization of the white matter tracts underpinning motor control in humans: an electrical stimulation study.
The somatotopic organization of the primary motor cortex is well documented. However, a possible somatotopy of the network involved in motor control, i.e., eliciting negative motor phenomena during electrostimulation, is unknown in humans, particularly at the subcortical level. Here, we performed electrical stimulation mapping in awake patients operated for gliomas, to study the distribution of the white matter tracts subserving movement control of the lower limb, upper limb(s), and speech. Eighteen patients underwent awake surgery for frontal low-grade gliomas, by using intraoperative subcortical electrostimulation mapping to search interference with movement of the leg, arm(s), and face. We assessed the negative motor responses and their distribution throughout the tracts located under premotor areas. The corresponding stimulation sites were reported on a standard brain template for visual analysis and between-subjects comparisons. During stimulation of the white matter underneath the dorsal premotor cortex and supplementary motor area, rostral to the corticospinal tracts, all patients experienced cessation of the movement of lower and upper limbs, of bimanual coordination, and/or speech. These subcortical sites were somatotopically distributed. Indeed, stimulation of the fibers from mesial to lateral directions and from posterior to anterior directions evoked arrest of movement of the lower limb (mesially and posteriorly), upper limb(s), and face/speech (laterally and anteriorly). There were no postoperative permanent deficits. This is the first evidence of a somatotopic organization of the white matter bundles underpinning movement control in humans. A better knowledge of the distribution of this motor control network may be helpful in neurosciences and neurosurgery.


Molecular cloning of tyrosine kinases in the early Xenopus embryo: identification of Eck-related genes expressed in cranial neural crest cells of the second (hyoid) arch.
Growth factors and their receptors play an important role in controlling cellular proliferation, migration, and differentiation during vertebrate embryogenesis. We have used the reverse transcription-polymerase chain reaction to survey the repertoire of receptor tyrosine kinases (TK) expressed during early embryogenesis of Xenopus laevis. Twelve distinct Xenopus TK cDNA classes were identified among a total of 352 cDNAs screened. A single TK cDNA class has been described previously and encodes the fibroblast growth factor receptor FGFR-A1. The remaining 11 TK cDNA classes appear to encode novel genes of the FGFR, platelet-derived growth factor receptor (PDGFR), Eph, Csk, Tyk2, and Klg subfamilies. By RNase protection assays, Xenopus TK mRNAs are rare transcripts (< 10(7) mRNA molecules/embryo), and are usually found to be expressed also maternally in the embryo. Most Xenopus TK genes examined by whole-mount in situ hybridization were expressed widely in tissues derived from multiple germ layers. Two Eck-related genes, however, were found to be restricted in their expression to neural crest of the second (hyoid) arch. Our findings are consistent with the proposed function of TKs in the regulation of specification and differentiation of embryonic tissues.


Pushing harder, pushing faster, minimizing interruptions… but falling short of 2010 cardiopulmonary resuscitation targets during in-hospital pediatric and adolescent resuscitation.
The objective of this study was to evaluate the effect of instituting the 2010 Basic Life Support Guidelines on in-hospital pediatric and adolescent cardiopulmonary resuscitation (CPR) quality. We hypothesized that quality would improve, but that targets for chest compression (CC) depth would be difficult to achieve. Prospective in-hospital observational study comparing CPR quality 24 months before and after release of the 2010 Guidelines. CPR recording/feedback-enabled defibrillators collected CPR data (rate (CC/min), depth (mm), CC fraction (CCF, %), leaning (%>2.5kg)). Audiovisual feedback for depth was: 2005, ≥38mm; 2010, ≥50mm; for rate: 2005, ≥90 and ≤120CC/min; 2010, ≥100 and ≤120CC/min. The primary outcome was average event depth compared with Student's t-test. 45 CPR events (25 before; 20 after) occurred, resulting in 1336 thirty-second epochs (909 before; 427 after). Compared to 2005, average event depth (50±13mm vs. 43±9mm; p=0.047), rate (113±11CC/min vs. 104±8CC/min; p<0.01), and CCF (0.94 [0.93, 0.96] vs. 0.9 [0.85, 0.94]; p=0.013) increased during 2010. CPR epochs during the 2010 period more likely to meet Guidelines for CCF (OR 1.7; CI95: 1.2-2.4; p<0.01), but less likely for rate (OR 0.23; CI95: 0.12-0.44; p<0.01), and depth (OR 0.31; CI95: 0.12-0.86; p=0.024). Institution of the 2010 Guidelines was associated with increased CC depth, rate, and CC fraction; yet, achieving 2010 targets for rate and depth was difficult.


MCAT is not required for in vitro polyketide synthesis in a minimal actinorhodin polyketide synthase from Streptomyces coelicolor.
It has been proposed that Streptomyces malonyl CoA: holo acyl carrier protein transacylases (MCATs) provide a link between fatty acid and polyketide biosynthesis. Two recent studies have provided evidence that the presence of MCAT is essential for polyketide synthesis to proceed in reconstituted minimal polyketide synthases (PKSs). In contrast to this, we previously showed that the holo acyl carrier proteins (ACPs) from type II PKSs are capable of catalytic self-malonylation in the presence of malonyl CoA, which suggests that MCAT might not be necessary for polyketide biosynthesis. We reconstituted a homologous actinorhodin (act) type II minimal PKS in vitro. When act holo-ACP is present in limiting concentrations, MCAT is required by the synthase complex in order for polyketide biosynthesis to proceed. When holo-ACP is present in excess, however, efficient polyketide synthesis proceeds without MCAT. The rate of polyketide production increases with holo-ACP concentration, but at low ACP concentration or equimolar AC:KS:CLF (KS, ketosynthase; CLF, chain length determining factor) concentrations this rate is significantly lower than expected, indicating that free holo-ACP is sequestered by the KS/CLF complex. The rate of polyketide biosynthesis is dictated by the ratio of holo-ACP to KS and CLF, as well as by the total protein concentration. There is no absolute requirement for MCAT in polyketide biosynthesis in vitro, although the role of MCAT during polyketide synthesis in vivo remains an open question. MCAT might be responsible for the rate enhancement of malonyl transfer at very low free holo-ACP concentrations or it could be required to catalyse the transfer of malonyl groups from malonyl CoA to sequestered holo-ACP.


Molecular characteristics of two laccase from the basidiomycete fungus Polyporus brumalis.
Two laccase cDNAs, pblac1 and pblac2, were cloned from a white-rot fungus strain, Polyporus brumalis (KFRI 20912). The cloned cDNAs consisted of 1,829 bp and 1,804 bp, and their open reading frames encoded proteins of 520 and 524 amino acids, with calculated molecular masses of approximately 55.9 kDa and 56 kDa, respectively. The deduced amino acid sequences of each protein showed 70% similarity. The copper binding regions were conserved in both proteins, as in other fungal laccases. RT-PCR analysis revealed that the transcript levels of the two laccases increased progressively in shallow stationary culture liquid medium. The transcript level of each laccase was induced when the fungus was exposed to di-butyl phthalate (DBP), suggesting that the two laccases are involved in DBP degradation. The overexpression of the pblac1 gene was derived by the promoter of a gene for glyceraldehyde-3-phosphate dehydrogenase, using a homologous system. The activity of laccase in the transformants was significantly higher than that of the wild type. The identification of these laccase cDNAs was a first step to characterize the molecular events related to the lignin degradation ability of this basidiomycetous fungus, as well as the degradation of many recalcitrant xenobiotics.


A case of solitary pulmonary metastasis of choroidal melanoma with an exceptionally long disease-free period.
Choroidal melanoma is the most common primary malignant intraocular tumor in adults. And its metastatic spread has been considered to be hematogenous with the liver being the most common site, and a solitary pulmonary metastasis without hepatic involvement is quite rare. We report the case of a 61-year-old woman with a solitary pulmonary metastasis from choroidal melanoma, which had been originally treated with enucleation 13 years ago. Video-assisted thoracoscopic surgery (VATS) right upper lobectomy was performed for the right upper lobe tumor, which was histopathologically confirmed to be metastatic choroidal melanoma. Our case is quite unusual in 2 points; firstly, a solitary pulmonary metastasis without hepatic involvement is rare. Secondly, this mode of metastasis with an exceptionally long disease-free period (13 years) is also rare. A pulmonary metastasis from choroidal melanoma is described with reference to relevant literature.


Diazoxide prevents H2O2-induced chondrocyte apoptosis and cartilage degeneration in a rat model of osteoarthritis by reducing endoplasmic reticulum stress.
Osteoarthritis (OA) is a common disease affecting elderly individuals. Its incidence rises sharply with age, and chondrocyte apoptosis plays a vital role in its pathogenesis. Diazoxide opens mitochondrial ATP-sensitive potassium (mitoKATP) channels and exerts multiple pharmacological effects, including reductions in blood pressure and blood sugar levels. It also exerts anti-apoptotic activities, but the cellular and molecular mechanisms by which diazoxide inhibits chondrocyte apoptosis are unknown, as is whether apoptosis is related to endoplasmic reticulum stress (ERS). In the present study, we explored the mechanism underlying the chondroprotective effect of diazoxide on hydrogen peroxide (H2O2)-stimulated chondrocyte apoptosis in rats with surgically induced OA. A cell counting kit-8 (CCK-8) assay showed that the viability of H2O2-stimulated chondrocytes was enhanced in a dose-dependent manner. However, at a concentration ≥400μM, diazoxide had other, negative effects. The protective effect of diazoxide in vitro included inhibition of the ERS response and of mitochondrial dysfunction induced by H2O2 stimulation. These responses were related to activation of the PERK1/2 and ERK1/2 signaling pathways; the prevention of chondrocyte apoptosis; the down-regulation of caspase-3, Bax, ATF-6 and C/EBP-homologous protein (CHOP) expression; and the up-regulation of Bcl-2 and Col II. In vivo, histological and immunohistochemical analyses of caspase-3 and CHOP expression revealed that diazoxide ameliorated cartilage degeneration in a rat model of OA, as revealed by histological and immunohistochemical analyses of caspase-3 and CHOP expression. Diazoxide suppressed H2O2-triggered chondrocyte apoptosis, and ameliorated cartilage degeneration, by inhibiting the development of ERS.


Changing the model of care delivery: nurses' perceptions of job satisfaction and care effectiveness.
To examine nurses' perceptions of job satisfaction, empowerment, and care effectiveness following a change from team to a modified total patient care (TPC) delivery model. Empirical data related to TPC is limited and inconclusive. Similarly, evidence demonstrating nurses' experience with change and restructuring is limited. A mixed method, longitudinal, descriptive design was used. Registered nurses and licenced practical nurses in two acute-care nursing units completed quantitative and qualitative surveys. Lewin's change theory provided the framework for the study. No significant change in job satisfaction was observed; however, it was less than optimal at all three time-periods. Nurses were committed to their jobs but relatively dissatisfied with their input into the goals and processes of the organization. Client care was perceived to be more effective under TPC. Job satisfaction remained consistent following the transition to TPC. However, nurses perceived that client care within the modified TPC model was more effective than in the previous model. Nursing administration must work collaboratively with nurses to improve processes in nursing practice that could enhance nurses' job satisfaction and improve client care delivery.


ACTH-secreting islet cell tumor: appearances on dynamic gadolinium-enhanced MRI.
Adrenocorticotropic hormone (ACTH) secreting islet cell tumors of the pancreas are extremely rare. A case of an ACTH producing islet cell tumor with multiple liver metastases is reported and the magnetic resonance imaging (MRI) findings are described.


[Birth weight, parturition and meat production in the dairy cow population].
Birth weight was found to be closely related to calving behaviour. With all variations in Friesian dairy cattle, rising beef yield proved to be achievable even along with medium birth weight, so that calving complications and resulting decline in reproduction can be avoided.


Prevalence of anastomotic leak and the impact of indocyanine green fluorescein imaging for evaluating blood flow in the gastric conduit following esophageal cancer surgery.
Anastomotic leak (AL) following esophagectomy for esophageal cancer (EC) remains an important cause of prolonged hospitalization and impaired quality of life. Recently, indocyanine green (ICG) fluorescein imaging has been used to evaluate the gastric conduit blood supply during EC surgery. Although several factors have been reported to be associated with AL, no studies have evaluated the relationships between risk factors for AL, including ICG fluorescein imaging. The purpose of this study was to investigate the risk factors associated with AL following esophagectomy and to evaluate the impact of ICG fluorescein imaging of the gastric conduit during EC surgery. One hundred and twenty patients undergoing esophagectomy with esophagogastric anastomosis for EC were enrolled in this retrospective study. Clinicopathological factors, preoperative laboratory variables, and surgical factors, including ICG fluorescence imaging, were analyzed to determine their association with AL. Univariate and multivariate logistic regression analysis was used to evaluate the impact of each of these factors on the incidence of AL. Among the 120 patients enrolled in the study, 10 (8.3%) developed AL. Univariate analysis demonstrated an increased risk of AL in patients with a high-neutrophil-to-lymphocyte ratio (p = 0.0500) and in patients who did not undergo ICG fluorescein imaging (p = 0.0057). Multivariate analysis revealed that the absence of ICG imaging was an independent risk factor for AL (p = 0.0098). Using ICG fluorescein imaging to evaluate blood flow in the gastric conduit might decrease the incidence of AL following EC surgery.


Silencing TRIM37 inhibits the proliferation and migration of non-small cell lung cancer cells.
Tripartite motif containing 37 (TRIM37), a member of the tripartite motif (TRIM) family, has been involved in the development and progression of several tumors. However, its role in non-small cell lung cancer (NSCLC) is still unclear. Therefore, the aim of this study was to investigate the expression pattern and role of TRIM37 in NSCLC. Our results showed that TRIM37 was highly expressed in human NSCLC cell lines. Knockdown of TRIM37 obviously inhibited the proliferation in vitro and xenografted tumor growth in vivo. Furthermore, knockdown of TRIM37 suppressed NSCLC cell migration and invasion by inhibiting the epithelial-mesenchymal transition (EMT) phenotype. Lastly, knockdown of TRIM37 greatly down-regulated the protein expression levels of β-catenin, cyclinD1 and c-myc in A549 cells. In conclusion, the present study revealed that TRIM37 plays an important role in the development and progression of NSCLC. Thus, TRIM37 may act a potential therapeutic target for treating NSCLC.


Effects of New-Generation Antiepileptic Drug Prophylaxis on Delayed Neurovascular Events After Aneurysmal Subarachnoid Hemorrhage.
Neuroelectric disruptions such as seizures and cortical spreading depolarization may contribute to the development of delayed cerebral ischemia (DCI) after aneurysmal subarachnoid hemorrhage (SAH). However, effects of antiepileptic drug prophylaxis on outcomes remain controversial in SAH. The authors investigated if prophylactic administration of new-generation antiepileptic drugs levetiracetam and perampanel was beneficial against delayed neurovascular events after SAH. This was a retrospective single-center cohort study of 121 consecutive SAH patients including 56 patients of admission World Federation of Neurological Surgeons grades IV - V who underwent aneurysmal obliteration within 72 h post-SAH from 2013 to 2021. Prophylactic antiepileptic drugs differed depending on the study terms: none (2013 - 2015), levetiracetam for patients at high risks of seizures (2016 - 2019), and perampanel for all patients (2020 - 2021). The 3rd term had the lowest occurrence of delayed cerebral microinfarction on diffusion-weighted magnetic resonance imaging, which was related to less development of DCI. Other outcome measures were similar among the 3 terms including incidences of angiographic vasospasm, computed tomography-detectable delayed cerebral infarction, seizures, and 3-month good outcomes (modified Rankin Scale 0 - 2). The present study suggests that prophylactic administration of levetiracetam and perampanel was not associated with worse outcomes and that perampanel may have the potential to reduce DCI by preventing microcirculatory disturbances after SAH. Further studies are warranted to investigate anti-DCI effects of a selective α-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor antagonist perampanel in SAH patients in a large-scale prospective study.


Acid resistance variability among isolates of Salmonella enterica serovar Typhimurium DT104.
Acid resistance could be an indicator of virulence since acid resistant strains are able to better survive the human stomach passage and in macrophages. We studied the acid resistance of several Salmonella Typhimurium DT104 strains isolated from food and humans and identified cellular parameters contributing to the enhanced acid resistance of these isolates. Acid resistance was tested in 37 Salmonella enterica Typhimurium serovar DT104 (S. Typhimurium DT104) strains. Acid adaptation at pH 5 followed by exposure for 2 h at pH 2.5 in the 27 human, nine nonhuman, and in two reference strains, revealed strong variation of acid survival. After 2 h at pH 2.5 six strains of S. Typhimurium DT104 were considered high acid resistant as they displayed a level of survival >10%, 14 strains were considered intermediate acid resistant (level of survival was <10% and >0.01%) and 19 strains were considered low acid resistant (level of survival <0.01%). Six strains were selected for further studies and proteomics revealed a relatively high amount of phase 2 flagellin in an acid-sensitive strain and a relatively high amount of the beta component of the H(+)/ATPase in an acid-resistant strain. Two strains were slightly more heat resistant possibly as the result of increased levels of DnaK or GroEL. A significant difference could be detected between human and food isolates regarding their acid resistance; all high acid-resistant strains were human isolates. S. Typhimurium DT104 is known for two decades and has a great impact on human health causing serious food-borne diseases. Our results suggest the existence of a positive correlation between acid resistance and pathogenicity in S. Typhimurium DT104 as all high acid-resistant strains were isolated from humans.


Genetic diversity and virulence characteristics of biofilm-producing uropathogenic Escherichia coli.
Uropathogenic E. coli (UPEC) strains exhibit different levels of biofilm formation that help adhesion of the bacteria to uroepithelial cells. We investigated the genetic diversity and virulence-associated genes (VAGs) of biofilm-producing UPEC. A collection of 107 biofilm-producing (BFP) UPEC strains isolated from patients with UTI in Iran were divided into three groups of strong, moderate, and weak BFPs after a quantitative microtiter plate assay, and the involvement of curli and cellulose in adhesion of the strains to T24 cell line was confirmed by the construction of csgD and yedQ mutants of two representative UPEC strains. BFP strains were tested for their genetic diversity, phylogenetic groups, and the presence of 15 VAGs. A significant decrease in adhesion of csgD and yedQ mutant strains confirmed the role of biofilm production in adhesion to uroepithelial cells. A high diversity was found among all three groups of strong (Di = 0.998), moderate (Di = 0.998), and weak (Di = 0.988) BFPs with majority of the strains belonging to phylogroups B2 (44.9%) and A (24.3%). Strong BFP strains carried significantly higher level papEF, hlyA, and iutA than other BFP groups. In contrast, the presence of fimH, focG, sfaS, set-1, and cvaC was more pronounced among weak BFP strains. There exists a high genetic diversity among the BFP strains with different VGA profiles. However, the high prevalence of phylogroup A among BFP strains suggests the fitness of commensal E. coli strains to cause UTI in this country.


A mutant of infectious Asia 1 serotype foot-and-mouth disease virus with the deletion of 10-amino-acid in the 3A protein.
Foot-and-mouth disease virus (FMDV) serotype Asia 1 is one of the most predominant endemic serotypes in China. Our previous study has generated a full-length cDNA clone (pBSAs) of an Asia 1 serotype FMDV (As1/CHA/05) isolated from bovine. To further study the properties of this virus, a mutant in the 3A region of the cDNA clone (pBSAs-3A10D), containing the deletion at position 93-102 of the 3A protein of As1/CHA/05, was generated by PCR and cloning. After synthesis of RNA in vitro and transfection, the recombinant rvAs-3A10D virus was recovered from BHK-21 cells. Characterization of the rvAs-3A10D revealed that the infectivity, immunoreactivity, and replication kinetics in BHK-21 and PK-15 cells and virulence in mice of the rvAs-3A10D were similar to that of its parent virus. Notably, while wild-type and recombinant viruses containing full-length sequence of the 3A replicated well in primary calf kidney cells, the mutant rvAs-3A10D failed to replicate in primary calf kidney cells in vitro. Apparently, the full-length sequence of 3A in As1/CHA/05 is a necessary component for its replication in calf kidney cells. The availability of this 3A deletion infectious cDNA clone may help in further investigating the virulent determinants of FMDV and potentially developing FMDV vaccines.


Risk Factors for Oral Human Papillomavirus Infection Among Young Men Who Have Sex With Men-2 Cities, United States, 2012-2014.
Men who have sex with men (MSM) are at risk for cancers attributable to human papillomavirus (HPV), including oropharyngeal cancer. Human papillomavirus vaccination is recommended for US MSM through age 26 years. Oral HPV infection is associated with oropharyngeal cancer. We determined oral HPV prevalence and risk factors among young MSM. The Young Men's HPV study enrolled MSM aged 18 through 26 years from clinics in Chicago and Los Angeles during 2012 to 2014. Participants self-reported demographics, sexual behaviors, vaccination and human immunodeficiency virus (HIV) status. Self-collected oral rinse specimens were tested for HPV DNA (37 types) by L1-consensus PCR. We calculated adjusted prevalence ratios (aPR) and 95% confidence intervals (CI) for risk factors associated with oral HPV among participants not previously vaccinated. Oral HPV was detected in 87 (9.4%) of 922; 9-valent vaccine types were detected in 37 (4.0%) of 922. Among HIV-positive participants, 17 (19.3%) of 88 had oral HPV detected. Oral HPV was more prevalent among those reporting first sex at 18 years of age or younger (aPR, 2.44; 95% CI, 1.16-5.12); HIV infection (aPR, 1.99; 95% CI, 1.14-3.48); greater than 5 sex partners within the past month (aPR, 1.93; 95% CI, 1.13-3.31); performing oral sex on greater than 5 partners within the last 3 months (aPR, 1.87; 95% CI, 1.12-3.13); and having greater than 5 male sex partners within the last 3 months (aPR, 1.76; 95% CI, 1.08-2.87). Only 454 (49.2%) of 922 were aware that HPV can cause oropharyngeal cancers. Many oral HPV infections were with types targeted by vaccination. Oral HPV infections were significantly associated with HIV and sexual behaviors. Fewer than half of participants were aware that HPV could cause oropharyngeal cancer.


Non-coding RNAs in Ciona intestinalis.
The analysis of animal genomes showed that only a minute part of their DNA codes for proteins. Recent experimental results agree, however, that a large fraction of these genomes are transcribed and hence are probably functional at the RNA level. A computational survey of vertebrate genomes has predicted thousands of previously unknown ncRNAs with evolutionarily conserved secondary structures. Extending these comparative studies beyond vertebrates is difficult, however, since most ncRNAs evolve quickly at the sequence level while conserving their characteristic secondarystructures. We report on a computational screen of structured ncRNAs in the urochordate lineage based on a comparison of the genomic data from Ciona intestinalis, Ciona savignyi and Oikopleura dioica. We predict >1000 ncRNAs with an evolutionarily conserved RNA secondary structure. Of these, about a quarter are located in introns of known protein coding sequences. A few RNA motifs can be identified as known RNAs, including approximately 300 tRNAs, some 100 snRNA genes and a few microRNAs and snoRNAs. www.bioinf.uni-leipzig.de/Publications/SUPPLEMENTS/05-008/


Transformation of dissolved organic matter in a novel groundwater recharge system with reclaimed water.
A novel process, enhanced direct injection-well recharge system (EnDir), can overcome the technical difficulties during the application of conventional surface spreading and has been developed to recharge groundwater with reclaimed water. In this study, removal and transformation of dissolved organic matter (DOM) in the system were investigated in laboratory-scale experiments. Results demonstrated that dissolved organic carbon and trihalomethane formation potential values could be reduced from 6.54 +/- 1.30 mg/L and 267.9 +/- 24.3 microg/L to 1.59 +/- 0.64 mg/L and 104.5 +/- 10.2 microg/L, respectively, as a result of DOM biodegradation in the aerobic short-term vadose soil treatment. Fluorescence spectra showed that aromatic protein-like substances and soluble microbial byproducts could be removed, to a great extent, in the soil system. Despite different removal efficiencies of DOM in different molecular weight fractions, the residual DOM was composed mainly of fulvic acid-like and humic acid-like substances, with molecular weights of 500 Da to 1 kDa.


Post Nissen syndrome.
The Nissen fundoplication is the most common anti-reflux operation performed. Gas bloat and inability to vomit after repair may be severe, but infrequently require reoperation; in contrast, other complications can be very debilitating or life-threatening. One hundred and sixteen patients who required reoperation for serious complications after Nissen repair are presented and classified according to the cause of the failed repair. Nissen complications resulted in recurrent reflux (86 per cent), severe dysphagia (60 per cent), esophageal dysmotility (48 per cent) and gastric perforation and fistualization (5 per cent). The "classic" Nissen fundoplication involves a blind 360 degree wrap, which includes the acid-producing parietal cell mass. The resulting pouch drains poorly and is, therefore, subject to gastric ulceration. Reoperation at our institution, using principally the Hill antireflux procedure, gave excellent or good results in 86 per cent, fair in 9 per cent and poor in 5 per cent. Three operative deaths (2.6 per cent) and one late death (0.9 per cent) occurred.


Therapeutic patient education and all-cause mortality in patients with chronic heart failure: a propensity analysis.
Meta-analyses of disease management programs have shown favorable effects in chronic heart failure. Therapeutic patient education forms an integral part of these programs and may influence mortality per se. We aimed to determine the relationship between therapeutic patient education applied in routine clinical practice and long-term mortality in chronic heart failure. From 2007 to 2010 (median follow-up: 27.2 months), heart failure patients were prospectively enrolled in a multicenter, 'real-world', French cohort by centers previously trained in therapeutic patient education. As educated and non-educated patient profiles were expected to differ, mortality was assessed using conventional multivariate analyses and analyses made on propensity-matched cohorts for the application of therapeutic patient education. Of the 3237 patients who participated in the study (67.5 years; 69.5% men), 2347 were educated (72.5%) and 890 were not educated (27.5%). Non-educated patients were older, more often female, and more severely diseased than educated patients. All-cause mortality was 17.3% in the educated group vs. 31.0% in the non-educated group (adjusted HR 0.70, 95% CI 0.58-0.84, P<.001). This association remained in paired groups after adjustment for all baseline covariates excluding (model 1) or including (model 2) cardiovascular medications and propensity score (HR 0.72, 95% CI 0.58-0.90, P=.003; HR 0.73, 95% CI 0.59-0.90, P=.004, respectively). In chronic heart failure, therapeutic patient education by trained healthcare professionals appears associated with lower all-cause mortality. These data may have important implications in terms of healthcare organization, because they suggest that therapeutic patient education should be developed in all types of cardiology centers.


ATR-IR and EPR spectroscopy for detecting the alterations in cortical synaptosomes induced by aluminium stress.
Aluminium (Al) is reported to promote free radical production, decrease the antioxidant enzyme status and disturb the enzyme activity involved in acetylcholine metabolism leading to cognitive dysfunction that are strongly associated with Alzheimer's disease (AD) pathogenesis. This work aimed at investigating the effect of Al-toxicity on synaptosomal membrane biophysical properties and lipid peroxidation during 65 days. We utilized ATR-IR spectroscopy to study the changes in membrane biochemical structure and biophysical properties of isolated rat cortical synaptosomes, and EPR spin trapping and labeling to follow NADPH oxidase activity and changes of membrane order parameter, respectively. The results showed increase in membrane fluidity and disorder in early 21d of AlCl3 treatment, while after 42d the membrane rigidity, packing, and order increased. The late (65d) an increase in the amount of unsaturated fatty acids, the accumulation of lipid peroxide end products, and ROS production were detected in rat cortex synaptosomes mediated by Al toxicity and oxidative stress (OS). A dramatic increase was also detected in Ca2+ level, synaptic membrane polarity, and EPR-detected order S-parameter. These outcomes strongly suggest that the synaptosomal membrane phospholipids underwent free radical attacks mediated by AlCl3 due to greater NOX activity, and the release of synaptic vesicles into synaptic cleft might be hindered. The adopted spectroscopic techniques have shed light on the biomolecular structure and membrane biophysical changes of isolated cortical synaptosomes for the first time, allowing researchers to move closer to a complete understanding of pathological tissues.


[The need for creating a patient medication record system and characteristics of its application in community pharmacies].
The objectives of this study were to explore the need for an information system of patient's medication record in pharmacies, where pharmacists could find all information about patient's condition, diseases and used medicines; to find out internet and information technologies usage in pharmacies. In order to evaluate the demand for a specialized information system the method of a questionnaire-based survey was used. Obtained answers indicated that an information system could be useful in pharmacies and information technologies would facilitate information exchange among different institutions directly responsible for the treatment of patients.


The role of 5-HT(2A) receptor antagonism in amphetamine-induced inhibition of A10 dopamine neurons in vitro.
The role of the 5-HT(2A) receptor in modulating amphetamine-induced inhibition of dopamine neuronal firing in A9 and A10 was investigated in rat midbrain slices. The antipsychotic drugs olanzapine and clozapine more potently reversed the amphetamine-induced inhibition in A10 neurons compared to A9 neurons. Risperidone (0.03 and 0.1 microM) reversed amphetamine-induced inhibition of firing activity similarly in A9 and A10. The dopamine D2 receptor antagonist (-)sulpiride (0.05 and 1 microM) reversed the amphetamine (10 microM)-induced inhibition of firing activity in A9 and A10 neurons. The selective 5-HT(2A) receptor antagonist MDL 100907 (0.05 microM), strongly enhanced the reversal of amphetamine-induced inhibition by (-)sulpiride in A10, but its effectiveness was much smaller in A9 dopamine neurons. We conclude that 5-HT(2A) receptor antagonism enhanced reversal of amphetamine-induced inhibition by dopamine D2 antagonism in A10, suggesting that dopamine D(2) receptor antagonism combined with 5-HT(2A) receptor antagonism may play a role in antipsychotic drug atypicality.


Determination of Np-237 by radiochemical neutron activation analysis combined with extraction chromatography.
A procedure for determination of 237Np, 238Pu, 239,240Pu and 241Pu in environmental samples is described. Neptunium-237 is determined using radiochemical neutron activation analysis with pre- and post-irradiation chemistry based on solvent extraction and extraction chromatography. 238Pu, 239,240Pu is determined using alpha spectrometry and 241Pu by liquid scintillation spectrometry. The vertical profiles of 237Np, 238Pu, 239,240Pu in bottom sediments from the Black Sea are presented.


Challenge of dose escalation in locally advanced unfavourable prostate cancer using HDR brachytherapy.
In men with locally advanced/high-risk prostate cancer, there is an ongoing challenge to achieve improved results. Dose escalation studies using three-dimensional conformal radiotherapy (3D-CRT) or intensity modulated radiation therapy (IMRT) have shown benefit particularly in the intermediate and poor risk groups of patients. Of concern, however, is the increase in documented rectal toxicity. High-dose rate brachytherapy (HDR-BT) as a boost in combination with external beam radiotherapy (EBRT) is an alternative strategy of dose escalation that can potentially achieve an even higher biological equivalent dose (BED) to the tumour. The results so far are very encouraging for men with poor prognosis disease. Moreover the technique is associated with very low rates of acute and late toxicity.


Wiring of bilirubin oxidases with redox polymers on gas diffusion electrodes for increased stability of self-powered biofuel cells-based glucose sensing.
A new redox polymer/bilirubin oxidase (BOD)-based gas diffusion electrode was designed to be implemented as the non-current and non-stability limiting biocathode in a glucose/O2 biofuel cell that acts as a self-powered glucose biosensor. For the proof-of-concept, a bioanode comprising the Os-complex modified redox polymer P(VI-co-AA)-[Os(bpy)2Cl]Cl and FAD-dependent glucose dehydrogenase to oxidize the analyte was used. In order to develop an optimal O2-reducing biocathode for the biofuel cell Mv-BOD as well as Bp-BOD and Mo-BOD have been tested in gas diffusion electrodes in direct electron transfer as well as in mediated electron transfer immobilized in the Os-complex modified redox polymer P(VI-co-AA)-[Os(diCl-bpy)2]Cl2. The resulting biofuel cell exhibits a glucose-dependent current and power output in the concentration region between 1 and 10 mM. To create a more realistic test environment, the performance and long-term stability of the biofuel cell-based self-powered glucose biosensor has been investigated in a flow-through cell design.


Endovascular repair of ruptured aberrant left subclavian artery with right aortic arch.
Association of a right-sided aortic arch with an aberrant left subclavian artery is rare. We present a case of successful endovascular repair of a ruptured Kommerell diverticulum associated with a right-sided aortic arch and aberrant left subclavian artery. We treated a 47-year-old woman with a diagnosis of ruptured aberrant left subclavian artery with thoracic endovascular stent-grafts. The descending aorta above Kommerell diverticulum was a reverse-tapered configuration. We managed the rather hostile neck with an extra-large Palmaz stent. A left carotid-to-subclavian bypass with an 8-mm Dacron graft was also performed to restore left arm perfusion and prevent vertebrobasilar insufficiency.


Establishment of seminars to improve the diagnostic accuracy and effectiveness of breast ultrasound.
To improve the ability of technicians and physicians to find and diagnose breast lesions in breast ultrasound screening. Seminars were organized for technicians and physicians engaged in breast ultrasound screening, and tests were carried out to evaluate the usefulness of the seminars. Each seminar lasted 2 days and comprised lectures and group activities. Pretests and post-tests conducted before and after each seminar, respectively, consisted of 100 questions: 50 about animated images, and 50 about static images. The tests required the participant to find lesions in animated images and estimate the probability of malignancy from static images. In the animated image tests, sensitivity was greater after the seminar, although specificity did not change significantly. In the static image tests, sensitivity increased, and a significant increase was also observed in the receiver operating curve analysis for degree of certainty in diagnosing cancer. The seminars improved the participants' ability to find and diagnose breast lesions during ultrasound screening.


Peripheral biomarkers for individualizing immunosuppression in transplantation--regulatory T cells.
Regulatory T cells have gained much interest in the research community over the last 15 years. Here we give a short overview on the phenotype of Tregs, described subsets and their mode of action. We also summarize findings obtained within the last 4 years studying peripheral Tregs numbers and their function in solid organ transplant patients. Of special interest for the transplant community is whether such an analysis can be used to 1) identify pre-transplant "high risk" patients likely to develop severe acute rejection episodes shortly after transplantation, 2) monitor drug-dependent reconstitution or changes of lymphocytes, 3) delineate and predict deterioration in graft function or rejections after transplantation and 4) identify potentially tolerant patients in whom immunosuppressive drugs can be safely withdrawn. The obtained results are very promising but mostly obtained by retrospective analysis on a limited number of patients. Thus, for the future collaborative efforts should be undertaken to initiate large prospective multi-center trials in which the relevance of analyzing Tregs can be further validated.


Adherence to oral antidiabetic therapy in a managed care organization: a comparison of monotherapy, combination therapy, and fixed-dose combination therapy.
Although medication adherence is one of the most important aspects of the management of diabetes mellitus, low rates of adherence have been documented. This study sought to examine medication adherence among patients with diabetes mellitus in a managed care organization who were receiving antidiabetic monotherapy (metformin or glyburide), combination therapy (metformin and glyburide), or fixed-dose combination therapy (glyburide/metformin). Medication adherence was evaluated through a retrospective database analysis of pharmacy claims. The adherence rate was defined as the sum of the days' supply of oral antidiabetic medication obtained by the patient during the follow-up period divided by the total number of days in the designated follow-up period (180 days). Health plan members were included in the analysis if they had an index pharmacy claim for an oral antidiabetic medication between August 1 and December 31, 2000, were continuously enrolled in the health plan, and were aged > or =18 years. A 6-month pre-index period was used to classify patients as newly treated or previously treated. Patients were grouped according to their medication-use patterns. After adjustment for potential confounding factors, including overall medication burden at index, there were no significant differences in adherence rates among 6502 newly treated patients receiving monotherapy, combination therapy, or fixed-dose combination therapy. Among the 1815 previously treated patients receiving glyburide or metformin monotherapy who required the addition of the alternative agent, resulting in combination therapy, adherence rates were significantly lower (54.0%; 95% CI, 0.52-0.55) than in the 105 patients receiving monotherapy who were switched to fixed-dose combination therapy (77.0%; 95% CI, 0.72-0.82). The 59 previously treated patients receiving combination therapy who were switched to fixed-dose combination therapy had a significant improvement in adherence after the switch (71.0% vs 87.0%; P < 0.001). In a managed care organization, previously treated patients receiving monotherapy with an oral antidiabetic medication who required additional therapy exhibited significantly greater adherence when they were switched to fixed-dose combination therapy compared with combination therapy. Patients receiving combination therapy who were switched to fixed-dose combination therapy exhibited significantly greater adherence after the switch.


Changing indications for enucleation and selected unusual pathologies.
To survey clinical indications for enucleations received within a single specialist ophthalmic pathology department over a 21-year period. Cases were identified from the departmental records and categorised by indication type as well as by year. Over time, there has been a decreasing trend in eyes removed for end-stage indications such as phthisis, blindness or pain, as well as a decrease in eyes removed following trauma. With the reducing number of eyes removed for end-stage disease and trauma, the majority of enucleations performed are now for malignancy. However, eyes are still removed for other indications. A selection of rarer pathologies is presented.


Multi-omics profiling reveals potential alterations in rheumatoid arthritis with different disease activity levels.
Rheumatoid arthritis (RA) is a chronic, systemic autoimmune inflammatory disease, the pathogenesis of which is not clear. Clinical remission, or decreased disease activity, is the aim of treatment for RA. However, our understanding of disease activity is inadequate, and clinical remission rates for RA are generally poor. In this study, we used multi-omics profiling to study potential alterations in rheumatoid arthritis with different disease activity levels. Fecal and plasma samples from 131 rheumatoid arthritis (RA) patients and 50 healthy subjects were collected for 16S rRNA sequencing, internally transcribed spacer (ITS) sequencing, and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The PBMCS were also collected for RNA sequencing and whole exome sequencing (WES). The disease groups, based on 28 joints and ESR (DAS28), were divided into DAS28L, DAS28M, and DAS28H groups. Three random forest models were constructed and verified with an external validation cohort of 93 subjects. Our findings revealed significant alterations in plasma metabolites and gut microbiota in RA patients with different disease activities. Moreover, plasma metabolites, especially lipid metabolites, demonstrated a significant correlation with the DAS28 score and also associations with gut bacteria and fungi. KEGG pathway enrichment analysis of plasma metabolites and RNA sequencing data demonstrated alterations in the lipid metabolic pathway in RA progression. Whole exome sequencing (WES) results have shown that non-synonymous single nucleotide variants (nsSNV) of the HLA-DRB1 and HLA-DRB5 gene locus were associated with the disease activity of RA. Furthermore, we developed a disease classifier based on plasma metabolites and gut microbiota that effectively discriminated RA patients with different disease activity in both the discovery cohort and the external validation cohort. Overall, our multi-omics analysis confirmed that RA patients with different disease activity were altered in plasma metabolites, gut microbiota composition, transcript levels, and DNA. Our study identified the relationship between gut microbiota and plasma metabolites and RA disease activity, which may provide a novel therapeutic direction for improving the clinical remission rate of RA.


New functions of epidermal growth factor: stimulation of capillary endothelial cell migration and matrix dependent proliferation.
The proliferative response of bovine retinal capillary endothelial cells to EGF is dependent upon attaching the cells to a matrix of fibronectin. Bovine capillary endothelial cells are also stimulated to actively migrate when exposed to EGF in vitro. These activities provide an explanation for the angiogenic properties of EGF in vivo. Capillary cell migration and proliferation are proposed as sensitive quantifiable bioassays to explore the functional domains of the EGF molecule. Studies on the inactivation of these properties of EGF by specific cleavage of the molecule with CNBr or proteases suggest that an intact loop composed in part by amino acid residues 20 to 31 is essential for at least some functions.


Indinavir acutely inhibits insulin-stimulated glucose disposal in humans: a randomized, placebo-controlled study.
Therapy with HIV protease inhibitors (PI) causes insulin resistance even in the absence of HIV infection, hyperlipidemia or changes in body composition. The mechanism of the effects on insulin action is unknown. In vitro studies suggest that PI selectively and rapidly inhibit the activity of the insulin-responsive glucose transporter GLUT-4. We hypothesized that a single dose of the PI indinavir resulting in therapeutic plasma concentrations would acutely decrease insulin-stimulated glucose disposal in healthy human volunteers. Randomized, double-blind, cross-over study comparing the effect of 1200 mg of orally administered indinavir and placebo on insulin-stimulated glucose disposal during a 180-min euglycemic, hyperinsulinemic clamp. Six healthy HIV-seronegative adult male volunteers were studied twice with 7 to 10 days between studies. There were no significant differences in baseline fasting body weight, or plasma glucose, insulin, lipid and lipoprotein levels between placebo- and indinavir-treated subjects. During steady-state (t60-180 min) insulin reached comparable levels (394 +/- 13 versus 390 +/- 11 pmol/l) and glucose was clamped at approximately 4.4 mmol/l under both conditions. The average maximum concentration of indinavir was 9.4 +/- 2.2 microM and the 2-h area under the curve was 13.5 +/- 3.1 microM.h. Insulin-stimulated glucose disposal per unit of insulin (M/I) decreased in all subjects from 14.1 +/- 1.2 to 9.2 +/- 0.8 mg/kg.min per microUI/ml (95% confidence interval for change, 3.7-6.1; P < 0.001) on indinavir (average decrease, 34.1 +/- 9.2%). The non-oxidative component of total glucose disposal (storage) decreased from 3.9 +/- 1.8 to 1.9 +/- 0.9 mg/kg.min (P < 0.01). Free fatty acid levels were not significantly different at baseline and were suppressed equally with insulin administration during both studies. A single dose of indinavir acutely decreases total and non-oxidative insulin-stimulated glucose disposal during a euglycemic, hyperinsulinemic clamp. Our data are compatible with the hypothesis that an acute effect of indinavir on glucose disposal in humans is mediated by a direct blockade of GLUT-4 transporters.


Neonatal piglet model of intraaortic balloon pumping: improved efficacy using echocardiographic timing.
Pediatric intraaortic balloon pumping (IABP) has met with little success because of technical difficulty in tracking rapid heart rates. This study was designed to evaluate the efficacy of M-mode echocardiography for IABP timing in a neonatal piglet model. Two groups of piglets underwent mitral valve avulsion to create a model of shock. Group 1 (n = 8; mean weight, 7.7+/-1.8 kg) underwent IABP timed with both the ascending aortic pressure and M-mode echocardiogram. Group 2 (n = 6; mean weight, 7.5+/-1.4 kg) underwent two separate periods of IABP: one with echocardiographic timing and the second using standard timing points from the femoral arterial pressure tracing and electrocardiogram. Measurements included ascending aortic flow, left anterior descending arterial flow, ascending aortic pressure, left atrial pressure, and heart rate. Mitral valve avulsion produced a shock model with a significant decrease in mean aortic pressure and aortic flow and a significant increase in left atrial pressure and heart rate. Compared with the shock state, IABP in group 1 animals resulted in a significant increase in aortic flow (353+/-152 versus 454+/-109 mL/min; p < 0.05) and a significant decrease in left atrial pressure (23+/-6 versus 17+/-7 mm Hg; p < 0.05). Group 2 animals with echocardiogram-timed IABP had significantly increased aortic flow (365+/-106 versus 458+/-107 mL/min; p < 0.05) and mean aortic pressure (43+/-11 versus 52+/-8 mm Hg; p < 0.05). However, standard-timed IABP failed to show any improvement. In piglets with rapid heart rates, echocardiogram-timed IABP results in increased aortic flow and pressure and decreased left atrial pressure compared with standard-timed IABP.


The Distance of the Common Peroneal Nerve to the Posterolateral Structures of the Knee.
The common peroneal nerve (CPN) is an important structure of the lower limb and is at risk of injury during total knee arthroplasty. The aim of this study was to use a tibial reference system to determine the position of the CPN relative to the knee center and popliteus. Two hundred consecutive knee magnetic resonance images at the level of a standard tibial arthroplasty cut were evaluated for (1) distance of the CPN from the posterolateral capsule; (2) angle of the CPN from the center of the tibial anteroposterior axis; and (3) location of CPN with respect to the popliteus. The mean distance between the CPN and the posterolateral joint capsule was 11.9 mm (range, 4.7-22.13 mm), which correlated positively with the medial-lateral axis of the tibia (Pearson correlation, 0.157; P = .026) and negatively with the angle of the nerve from the midline (Pearson correlation, -0.237, P = .001). The mean angle of the nerve from the midline was 42.2° (range, 25.0°-64.0°). In 116 knees (58%), the CPN was in line with the popliteus from the center of the knee, in 69 knees (34.5%) the CPN was lateral to the popliteus, and in 15 knees (7.5%), the CPN was medial to the popliteus. A danger zone was identified as between 29.95° and 54.57° from the anteroposterior axis. The CPN is at risk during total knee arthroplasty. This study describes a method to help predict the location of the CPN intraoperatively and therefore avoid direct injury.


Pentamidine-Loaded Lipid and Polymer Nanocarriers as Tunable Anticancer Drug Delivery Systems.
Initially developed as a synthetic analogue of insulin, pentamidine (PTM) is an antimicrobial drug that has recently shown in vitro and in vivo anticancer activity. Nevertheless, systemic administration of PTM causes severe side effects, especially nephrotoxicity. Here we propose the association of PTM to different biocompatible nanosystems in order to compare the physicochemical characteristics of the loaded nanocarriers and their influence on the drug cytotoxicity toward cancer cells. In particular, PTM (as free base or with different counterions) was encapsulated into liposomes and poly(lactide-co-glycolide) (PLGA) nanoparticles and all the formulations have been deeply characterized concerning mean diameter, polydispersity index, zeta potential, stability, morphology, PTM loading, and drug release profile. The anticancer activity was evaluated on a human ovarian cancer cell line over 72 h. Results showed that PTM is efficiently loaded into liposomes with a transmembrane citrate or sulfate gradient; concerning PLGA nanoparticles, important association occurred, thanks to ionic interactions between the drug and the polymer. The in vitro studies confirmed the anticancer activity of PTM, which was gradually released with different profiles depending on the drug form and the nanocarrier structure.


Interaction mechanism and spatial effect of cross-regional haze pollution based on a multisectoral economy-energy-environment (3E) model and the evidence from China.
The transboundary characteristics and multisectoral factor interaction mechanism of haze pollution have aroused widespread attention but remain understudied. This article proposes a comprehensive conceptual model that clarifies regional haze pollution, further establishes a theoretical framework on a cross-regional, multisectoral economy-energy-environment (3E) system, and attempts to empirically investigate the spatial effect and interaction mechanism employing a spatial-econometrics model based on China's province-level regions. The results demonstrate that (1) regional haze pollution is a transboundary atmospheric state formed by the accumulation and agglomeration of various emission pollutants; moreover, there is a "snowball" effect and a spatial spillover effect. (2) The formation and evolution of haze pollution are driven by the multisectoral factors of 3E system interaction, and the findings still hold after theoretical and empirical analysis and robustness tests. (3) Significant spatial autocorrelation exists for the 3E factors, presenting different clustering modes with a dynamic spatiotemporal evolution, particularly in the high-high (H-H) mode and low-low (L-L) mode. (4) Significant heterogeneous impacts of economic and energy factors on haze pollution are identified, namely, an inverted "U-shaped" relationship and a positive linear association, respectively. Further spatial analysis demonstrates a strong spatial spillover and obvious path dependence among local and neighboring regions. Policymakers are advised to consider multisectoral 3E system interaction and cross-regional collaboration. Integr Environ Assess Manag 2023;19:1525-1543. © 2023 SETAC.


Detection of receptors for the Fc region of IgG on streptococci.
A semi-quantitative absorption test to measure Fc-reactive proteins on the surface of streptococci is described. The ability of bacteria to remove intact IgG in the presence of an equimolar amount of F(ab')2 fragments was used to identify streptococci with Fc-reactive proteins on their surface. This method was found to be more objective, reproducible and quantitative than the hemagglutination and 125I-labeled IgG binding methods currently in use. Methods for detection of secreted Fc-reactive materials with protein A-like reactivity are also described.


Mammalian DNA2 helicase/nuclease cleaves G-quadruplex DNA and is required for telomere integrity.
Efficient and faithful replication of telomeric DNA is critical for maintaining genome integrity. The G-quadruplex (G4) structure arising in the repetitive TTAGGG sequence is thought to stall replication forks, impairing efficient telomere replication and leading to telomere instabilities. However, pathways modulating telomeric G4 are poorly understood, and it is unclear whether defects in these pathways contribute to genome instabilities in vivo. Here, we report that mammalian DNA2 helicase/nuclease recognizes and cleaves telomeric G4 in vitro. Consistent with DNA2's role in removing G4, DNA2 deficiency in mouse cells leads to telomere replication defects, elevating the levels of fragile telomeres (FTs) and sister telomere associations (STAs). Such telomere defects are enhanced by stabilizers of G4. Moreover, DNA2 deficiency induces telomere DNA damage and chromosome segregation errors, resulting in tetraploidy and aneuploidy. Consequently, DNA2-deficient mice develop aneuploidy-associated cancers containing dysfunctional telomeres. Collectively, our genetic, cytological, and biochemical results suggest that mammalian DNA2 reduces replication stress at telomeres, thereby preserving genome stability and suppressing cancer development, and that this may involve, at least in part, nucleolytic processing of telomeric G4.


IFNG +874 A/T is associated with acute lymphoblastic leukemia in Mexican Mestizos.
Acute lymphoblastic leukemia (ALL), the most common type of cancer in children worldwide, has one of the highest incidence rates in Mexico. It is a multifactorial disease and different cytokine single nucleotide polymorphisms (SNP), have been associated with ALL expression. Few studies have been published analyzing IFNG +874 T/A and IL2 -330 G/T in this type of leukemia. These SNPs are involved in high or low expression, and are central to cellular immunity, influencing greatly tumor growth. The purpose of this work was to explore the association of IFNG +874 A/T (rs2430561) and IL2 -330 G/T (rs2069762) SNPs with ALL susceptibility and/or protection in 488 Mexican Mestizos patients, as compared to 950 Mexican Mestizo healthy controls. The results demonstrated that IFNG +874 T allele (pc = 0.00004, OR = 0.673) and the TT genotype (pc = 0.00015, OR = 0.349), protect against ALL expression with no specific gender association; however, the TT homozygote genotype (vs. TA+AA) seems more protective in males (pc = 0.00683). IL2 -330 G/T does not contribute to the development of ALL. In healthy Mexicans, the most common genotypes for IL2 and IFNG, are the low cytokine producers, suggesting that the genetic background in this ethnic group, may be partly responsible for the high incidence of ALL. These results show for the first time in Mexicans, the relevant role that IFNG SNP has in the genetic etiology of ALL. Thus, a large group of patients belonging to different ethnicities will be very helpful to study in order to demonstrate if these SNPs contribute to the genetic etiology of ALL, as shown here in Mexican Mestizos.


Screening diabetic patients at risk for foot ulceration. A multi-centre hospital-based study in France.
To determine the prevalence of risk factors for diabetic foot ulceration in diabetic patients free of active pedal ulceration in a hospital setting. In sixteen French diabetology centres, a survey was conducted on a given day in all diabetic people attending the units, both as in- or out-patients. 664 patients were evaluated: 105 had an active foot ulcer and were excluded from the analysis as were four other patients due to lack of reliable data. From the 555 assessable patients, 40 (7.2%) had a history of foot ulcer or lower-limb amputation. Sensory neuropathy with loss of protective sensation, as measured by the 5.07 (10 g) Semmes-Weinstein monofilament testing, was present in 27.1% of patients, whereas 17% had a peripheral arterial disease mainly based on the clinical examination. On addition, foot deformities were found in 117 patients (21.1%). According to the classification system of the International Working Group on the Diabetic Foot, 72.8% of patients were at low-risk for pedal ulceration (grade 0) and 17,5% were in the higher-risk groups (grade 2 & 3). If patients with isolated peripheral arterial disease were considered as a separate risk group (as was those with isolated neuropathy), percentage of low-risk patients decreased to 65.6%. There was a clear trend between the increasing severity of the staging and age, duration of diabetes, prevalence of nephropathy and retinopathy. Prevalence of risk factors for foot ulceration is rather high in a hospital-based diabetic population, emphasising the need for implementing screening and preventive strategies to decrease the burden of diabetic foot problems and to improve the quality of life for people with diabetes.


A comparative evaluation of different types of microbial electrolysis desalination cells for malic acid production.
The aim of this study was to investigate different microbial electrolysis desalination cells for malic acid production. The systems included microbial electrolysis desalination and chemical-production cell (MEDCC), microbial electrolysis desalination cell (MEDC) with bipolar membrane and anion exchange membrane (BP-A MEDC), MEDC with bipolar membrane and cation exchange membrane (BP-C MEDC), and modified microbial desalination cell (M-MDC). The microbial electrolysis desalination cells performed differently in terms of malic acid production and energy consumption. The MEDCC performed best with the highest malic acid production rate (18.4 ± 0.6 mmol/Lh) and the lowest energy consumption (0.35 ± 0.14 kWh/kg). The best performance of MEDCC was attributable to the neutral pH condition in the anode chamber, the lowest internal resistance, and the highest Geobacter percentage of the anode biofilm population among all the reactors.


Inducing and Inhibiting Autophagy to Investigate Its Interactions with MIF.
MIF has been described to be associated with autophagy in a number of studies, but the full nature of their association is not yet clear. However, the unprecedented interest in autophagy in recent times has generated a number of tools and techniques for its study. Here, we present protocols for studying the interactions between MIF and autophagy, including for the induction and inhibition of autophagy and measuring autophagosome biogenesis and maturation.


Highly potent anti-inflammatory, analgesic and antioxidant activities of 3,5-disubstituted tetrahydro-2H-1,3,5-thiadiazine thiones.
Four series of tetrahydro-2H-1,3,5-thiadiazine-2-thiones (series A and B including two novel enantiopure isomers), tetrahydro-2H-1,3,5-thiadiazine-6-thiones (series C) and N-3 ester derivatives of tetrahydro-2H-1,3,5-thiadiazine-6-thiones (series D) were synthesized and evaluated for their anti-inflammatory, analgesic and anti-oxidant activities. These THTT analogues specially series D were first time examined for their in vitro anti-inflammatory, in vivo analgesic and anti-oxidant activities. Among them lipophilic compounds (series B and D) were found to be highly active for anti-inflammatory evaluation with IC50 values between 5.1-16.9 and 4.1-32.4 μM, respectively when compared with the standard drug ibuprofen IC50 = 11.2 μM. The structure-activity relationship exposed the importance of lipophilic substituents especially ester and n-propyl group for inhibition of inflammation. The molecular docking studies demonstrated that all the active analogues of THTT have notable binding relations with Arg120 of the active sites of COX-1 enzyme either through CS moiety of the THTT nucleus or with COO attached at N-3 of THTT nucleus. In vivo analgesic activity of the selected THTT compounds 14, 17, 18, 19 (series B) and 28 (series D) were also carried out by acetic acid-induced writhing procedure. The compound 28 showed significant anti-nociceptive/analgesic activity at the oral dose of 5 mg/kg body weight with the percent protection (32.05 %) when compared with standard indomethacin at 10 mg/kg (48.83 %). Additionally, these compounds demonstrated the moderate level of antioxidant potential with IC50 values in the range of 60.9 to 93.6 μM (standard butylated hyroxyanisole; IC50 = 44.2 μM). These results indicated that this class of heterocyclic compounds may be a template specially to design better anti-inflammatory and analgesic agents.


Defensive mobilization in specific phobia: fear specificity, negative affectivity, and diagnostic prominence.
Understanding of exaggerated responsivity in specific phobia-its physiology and neural mediators-has advanced considerably. However, despite strong phenotypic evidence that prominence of specific phobia relative to co-occurring conditions (i.e., principal versus nonprincipal disorder) is associated with dramatic differences in subjective distress, there is yet no consideration of such comorbidity issues on objective defensive reactivity. A community sample of specific phobia (n = 74 principal; n = 86 nonprincipal) and control (n = 76) participants imagined threatening and neutral events while acoustic startle probes were presented and eyeblinks (orbicularis occuli) recorded. Changes in heart rate, skin conductance level, and facial expressivity were also measured. Principal specific phobia patients far exceeded control participants in startle reflex and autonomic reactivity during idiographic fear imagery. Distinguishing between single and multiple phobias within principal phobia and comparing these with nonprincipal phobia revealed a continuum of decreasing defensive mobilization: single patients were strongly reactive, multiple patients were intermediate, and nonprincipal patients were attenuated-the inverse of measures of pervasive anxiety and dysphoria (i.e., negative affectivity). Further, as more disorders supplanted specific phobia from principal disorder, overall defensive mobilization was systematically more impaired. The exaggerated responsivity characteristic of specific phobia is limited to those patients for whom circumscribed fear is the most impairing condition and coincident with little additional affective psychopathology. As specific phobia is superseded in severity by broad and chronic negative affectivity, defensive reactivity progressively diminishes. Focal fears may still be clinically significant but not reflected in objective defensive mobilization.


The complex relationship between genotype, pathology and phenotype in familial dementia.
Causative genes involved in familial forms of dementias, including Alzheimer's disease (AD), frontotemporal lobar degeneration (FTLD) and dementia with Lewy bodies (DLB), as well as amyotrophic lateral sclerosis and prion diseases where dementia is present as a significant clinical feature, are associated with distinct proteinopathies. This review summarizes the relationship between known genetic determinants of these dementia syndromes and variations in key neuropathological proteins in terms of three types of heterogeneity: (i) Locus Heterogeneity, whereby mutations in different genes cause a similar proteinopathy, as exemplified by mutations in APP, PSEN1 and PSEN2 leading to AD neuropathology; (ii) Allelic Heterogeneity, whereby different mutations in the same gene lead to different proteinopathies or neuropathological severity, as exemplified by different mutations in MAPT and PRNP giving rise to protein species that differ in their biochemistry and affected cell types; and (iii) Phenotypic Heterogeneity, where identical gene mutations lead to different proteinopathies, as exemplified by LRRK2 p.G2019S being associated with variable Lewy body presence and alternative AD neuropathology or FTLD-tau. Of note, the perceived homogeneity in histologic phenotypes may arise from laboratory-specific assessment protocols which can differ in the panel of proteins screened. Finally, the understanding of the complex relationship between genotype and phenotype in dementia families is highly relevant in terms of therapeutic strategies which range from targeting specific genes, to a broader strategy of targeting a downstream, common biochemical problem that leads to the histopathology.


How are declarations of interest working? A cross-sectional study in declarations of interest in healthcare practice in Scotland and England in 2020/2021.
To understand arrangements for healthcare organisations' declarations of staff interest in Scotland and England in the context of current recommendations. Cross-sectional study of a random selection of National Health Service (NHS) hospital registers of interest by two independent observers in England, all NHS Boards in Scotland and a random selection of Clinical Commissioning Groups (CCGs) in England. NHS Trusts in England (NHSE), NHS Boards in Scotland, CCGs in England, and private healthcare organisations. Registers of declarations of interest published in a random sample of 67 of 217 NHS Trusts, a random sample of 15 CCGs of in England, registers held by all 14 NHS Scotland Boards and a purposeful selection of private hospitals/clinics in the UK. Adherence to NHSE guidelines on declarations of interests, and comparison in Scotland. 76% of registers published by Trusts did not routinely include all declaration of interest categories recommended by NHS England. In NHS Scotland only 14% of Boards published staff registers of interest. Of these employee registers (most obtained under Freedom of Information), 27% contained substantial retractions. In England, 96% of CCGs published a Gifts and Hospitality register, with 67% of CCG staff declaration templates and 53% of governor registers containing full standard NHS England declaration categories. Single organisations often held multiple registers lacking enough information to interpret them. Only 35% of NHS Trust registers were organised to enable searching. None of the private sector organisations studied published a comparable declarations of interest register. Despite efforts, the current system of declarations frequently lacks ability to meaningfully obtain complete healthcare professionals' declaration of interests.


Medicolegal issues in cluster headache.
This paper identifies legal issues of relevance to the diagnosis and treatment of cluster headache, including areas of actual and potential malpractice liability. Legal topics that are relevant to cluster headache can be divided into five categories: diagnostic-related issues, risks inherent in the disease process, prescribing and treatment-related problems, research-related issues, and disability determination.


Cytoarchitecture and cortical connections of the anterior cingulate and adjacent somatomotor fields in the rhesus monkey.
The cytoarchitecture and cortical connections of the anterior cingulate, medial and dorsal premotor, and precentral region are investigated using the Nissl and NeuN staining methods and the fluorescent retrograde tract tracing technique. There is a gradual stepwise laminar change in the cytoarchitectonic organization from the proisocortical anterior cingulate region, through the lower and upper banks of the cingulate sulcus, to the dorsolateral isocortical premotor and precentral motor regions of the frontal lobe. These changes are characterized by a gradational emphasis on the lower stratum layers (V and VI) in the proisocortical cingulate region to the upper stratum layers (II and III) in the premotor and precentral motor region. This is accompanied by a progressive widening of layers III and VI, a poorly delineated border between layers III and V and a sequential increase in the size of layer V neurons culminating in the presence of giant Betz cells in the precentral motor region. The overall patterns of corticocortical connections paralleled the sequential changes in cytoarchitectonic organization. The proisocortical areas have connections with cingulate motor, supplementary motor, premotor and precentral motor areas on the one hand and have widespread connections with the frontal, parietal, temporal and multimodal association cortex and limbic regions on the other. The dorsal premotor areas have connections with the proisocortical areas including cingulate motor areas and supplementary motor area on the one hand, and premotor and precentral motor cortex on the other. Additionally, this region has significant connections with posterior parietal cortex and limited connections with prefrontal, limbic and multimodal regions. The precentral motor cortex also has connections with the proisocortical areas and premotor areas. Its other connections are limited to the somatosensory regions of the parietal lobe. Since the isocortical motor areas on the dorsal convexity mediate voluntary motor function, their close connectional relationship with the cingulate areas form a pivotal limbic-motor interface that could provide critical sources of cognitive, emotional and motivational influence on complex motor function.


Sex and Age Differences in Patients With Unstable Angina Pectoris: A Single-Center Retrospective Study.
Sex and age may affect the pathogenesis of coronary heart disease, such as cardiovascular risk factors, treatment and prognosis, but this information is not well known. This was a single-center retrospective cohort study. Patients with unstable angina pectoris between January 2013 and June 2018 were included and stratified into 4 age groups (<55, 55-64, 65-74 and ≥75 years). The cardiovascular risk factors profile, treatment and in-hospital prognosis differences by sex and age were explored. This study included 5,908 patients (2,198 women). The women were older than the men (mean age 67 vs. 62 years). Approximately 2 of 3 patients had ≥3 cardiovascular risk factors. Men were more likely to be smokers, and women had a higher level of total cholesterol, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol. Hypertension, diabetes and chronic kidney disease were more frequent in women ≥ 65 years old than in similarly aged men. Men and women less than 65 years of age had more frequent family history of coronary heart disease, higher body mass index, higher fasting plasma glucose, and higher lipid levels, especially for patients <55 years of age. More women tended to receive medical therapy than men (51.6% vs. 42.8%, P < 0.01). The overall incidence of in-hospital major adverse cardiovascular events was higher in men than in women (4.1% vs. 2.6%, P < 0.05), whereas there was no sex difference in the in-hospital cardiac mortality (0.2% vs. 0.2%, P > 0.05). Women had higher cholesterol levels, and were less likely to undergo revascularization therapy than similarly aged men, and elderly women had a higher prevalence of hypertension, diabetes, and chronic kidney disease than elderly men. In-hospital major adverse cardiovascular events were lower in women than in men; however, there was no sex difference in the in-hospital cardiac mortality.


Transcriptional profiling of Vero E6 cells over-expressing SARS-CoV S2 subunit: insights on viral regulation of apoptosis and proliferation.
We have previously demonstrated that over-expression of spike protein (S) of severe acute respiratory syndrome coronavirus (SARS-CoV) or its C-terminal subunit (S2) is sufficient to induce apoptosis in vitro. To further investigate the possible roles of S2 in SARS-CoV-induced apoptosis and pathogenesis of SARS, we characterized the host expression profiles induced upon S2 over-expression in Vero E6 cells by oligonucleotide microarray analysis. Possible activation of mitochondrial apoptotic pathway in S2 expressing cells was suggested, as evidenced by the up-regulation of cytochrome c and down-regulation of the Bcl-2 family anti-apoptotic members. Inhibition of Bcl-2-related anti-apoptotic pathway was further supported by the diminution of S2-induced apoptosis in Vero E6 cells over-expressing Bcl-xL. In addition, modulation of CCN E2 and CDKN 1A implied the possible control of cell cycle arrest at G1/S phase. This study is expected to extend our understanding on the pathogenesis of SARS at a molecular level.


Identifying the Profile of Helicobacter pylori-Negative Gastric Cancers: A Case-Only Analysis within the Stomach Cancer Pooling (StoP) Project.
The prevalence of Helicobacter pylori-negative gastric cancer (HpNGC) can be as low as 1%, when infection is assessed using more sensitive tests or considering the presence of gastric atrophy. HpNGC may share a high-risk profile contributing to the occurrence of cancer in the absence of infection. We estimated the proportion of HpNGC, using different criteria to define infection status, and compared HpNGC and positive cases regarding gastric cancer risk factors. Cases from 12 studies from the Stomach cancer Pooling (StoP) Project providing data on H. pylori infection status determined by serologic test were included. HpNGC was reclassified as positive (eight studies) when cases presented CagA markers (four studies), gastric atrophy (six studies), or advanced stage at diagnosis (three studies), and were compared with positive cases. A two-stage approach (random-effects models) was used to pool study-specific prevalence and adjusted odds ratios (OR). Among non-cardia cases, the pooled prevalence of HpNGC was 22.4% (n = 166/853) and decreased to 7.0% (n = 55) when considering CagA status; estimates for all criteria were 21.8% (n = 276/1,325) and 6.6% (n = 97), respectively. HpNGC had a family history of gastric cancer more often [OR = 2.18; 95% confidence interval (CI), 1.03-4.61] and were current smokers (OR = 2.16; 95% CI, 0.52-9.02). This study found a low prevalence of HpNGC, who are more likely to have a family history of gastric cancer in first-degree relatives. Our results support that H. pylori infection is present in most non-cardia gastric cancers, and suggest that HpNGC may have distinct patterns of exposure to other risk factors.


Superoxide fluxes limit nitric oxide-induced signaling.
Independently, superoxide (O2-) and nitric oxide (NO) are biologically important signaling molecules. When co-generated, these radicals react rapidly to form powerful oxidizing and nitrating intermediates. Although this reaction was once thought to be solely cytotoxic, herein we demonstrate using MCF7, macrophage, and endothelial cells that when nanomolar levels of NO and O2- were produced concomitantly, the effective NO concentration was established by the relative fluxes of these two radicals. Differential regulation of sGC, pERK, HIF-1alpha, and p53 were used as biological dosimeters for NO concentration. Introduction of intracellular- or extracellular-generated O2- during NO generation resulted in a concomitant increase in oxidative intermediates with a decrease in steady-state NO concentrations and a proportional reduction in the levels of sGC, ERK, HIF-1alpha, and p53 regulation. NO responses were restored by addition of SOD. The intermediates formed from the reactions of NO with O2- were non-toxic, did not form 3-nitrotyrosine, nor did they elicit any signal transduction responses. H2O2 in bolus or generated from the dismutation of O2- by SOD, was cytotoxic at high concentrations and activated p53 independent of NO. This effect was completely inhibited by catalase, suppressed by NO, and exacerbated by intracellular catalase inhibition. We conclude that the reaction of O2- with NO is an important regulatory mechanism, which modulates signaling pathways by limiting steady-state levels of NO and preventing H2O2 formation from O2-.


Neuromuscular patterns of stereotypic hindlimb behaviors in the first two postnatal months. I. Stepping in normal kittens.
Neuromuscular patterns associated with the development of hindlimb stepping behaviors were studied from birth to postnatal day 60 in normal kittens. Hindlimb muscles were chronically implanted with EMG electrodes at birth to characterize interlimb coordination and intralimb synergies during development of overground and treadmill stepping. Airstepping was also examined but seldom occurred after the second postnatal week. All kittens performed stepping under each condition, including weight-supported stepping, by postnatal day 3. The number of sequential steps on the treadmill and overground increased with age and cycle periods decreased. At onset, stepping behaviors were characterized by adult-like EMG patterns. Interlimb coordination was typified by alternating extensor bursts of similar duration. Extensors at the knee and ankle were coactive during the stance phase, and extensor burst durations were strongly correlated with the cycle periods over a wide range of stepping frequency. Ankle flexor and extensor muscles were reciprocally active during postural tremor, bouts of airstepping, and weight-supported steps on the treadmill and overground. The duration of the reciprocal flexor bust did not vary with cycle period or age. Observations of stepping behaviors and adult-like EMG patterns during initial postnatal development were contingent on optimal testing conditions. Taken together, the data suggest that pattern-generating circuits for regulating interlimb coordination and intralimb muscle synergies are potentially functional prior to the normal ontogenetic onset of locomotion. Perhaps the prolonged postnatal development of locomotion reflects the time required to establish adaptive mechanisms, such as postural control and agility, rather than spinal pattern-generating circuits for locomotion.


What are we discarding during the life cycle of a building? Case studies of social housing in Andalusia, Spain.
The paper evaluates for the first time the embodied impact in CDW during the buildings life cycle by means of the bill of quantities of construction projects. The main objective is to be able to predict the future CDW to be generated by a project in the design stage, by means of the bill of quantities of the urbanization, construction, renovation, rehabilitation and demolition projects. The tools already in place for cost control can be used as an instrument for the introduction of sustainability considerations in construction projects. The methodology proposes a connection between the different stages of a building's life cycle, more precisely its budget. The latter is linked to other future budgets for building renovations or retrofitting projects. The result shows that urbanization and demolition generate 90% of CDW, the former is caused by earthworks and the latter is due to the elimination of all building materials. The building is removed 1.3 times, in terms of material weight, energy and water. Finally, traditional models for economic control and waste management in construction projects can be the vector which introduce environmental assessment through the building life cycle.


Separation of chelating agents as copper complexes by capillary zone electrophoresis using quaternary ammonium bromides as additives in N-methylformamide.
This study presents the use of quaternary ammonium bromides as additives in N-methylformamide (NMF) for the separation and quantification of chelating agents as copper complexes by capillary zone electrophoresis (CZE). The new quaternary ammonium bromides were synthesized in our laboratory and used for the first time for CZE applications performed in NMF media. The methods were developed and optimized for determination of six chelating agents (trans-1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid (CDTA), diethylenetriaminepentaacetic acid (DTPA), ethylenediaminetetraacetic acid (EDTA), N-(2-hydroxyethyl)ethylenediamine-N,N',N'-triacetic acid (HEDTA), nitrilotriacetic acid (NTA) and triethylenetetraaminehexaacetic acid (TTHA)) as copper complexes. Among the tested electrolyte additives in NMF media (pH(app) 10.2) dimethyldioctylammonium bromide (DMDOAB), dimethyldinonylammonium bromide (DMDNAB) and dimethyldidecylammonium bromide (DMDDAB), at a concentration of 20 mmol L(-1) improved the separation of the copper complexes. The optimized methods require only 12 min for one analysis, and the detection limits for copper complexes of DMDNAB, the best-performing additive, were < or =24 micromol L(-1). Relative standard deviations (R.S.D.) for migration times were < or =2.5, < or =2.1, < or =3.1% and for peak areas, < or =3.1, < or =3.0, < or =3.2% for DMDOAB, DMDNAB and DMDDAB used as additives, respectively. All three methods were successfully applied to the analysis of natural and wastewater samples. No matrix effects from these samples were observed. The interaction between quaternary ammonium bromides and copper complexes is discussed.


Inhalation of nitric oxide as a treatment of pulmonary hypertension in congenital diaphragmatic hernia.
Congenital diaphragmatic hernia (CDH) still has a mortality risk of around 40%. The concomitant pulmonary hypoplasia and the persistent pulmonary hypertension are of major prognostic importance. The use of a selective pulmonary vasodilator may revert this vicious circle that is fatal to many children. Inhalation of nitric oxide (NO) has been suggested as a rational treatment of this condition. The authors report three cases of high-risk infants with CDH where NO was used successfully. It is concluded that hypoxemia in CDH can be treated successfully with NO inhalation when conventional treatment fails, and it may in some cases prove to be an alternative to ECMO.


[Educational plans as control instruments in youth care in the Netherlands].
The significance, development, and different forms of educational plans and the special role of mentors and case managers in youth care in the Netherlands are presented.


Heat resistance in extended-spectrum beta-lactamase-producing Escherichia coli may favor environmental survival in a hospital setting.
Nosocomial infections caused by extended-spectrum β-lactamase (ESBL)-producing Escherichia coli are a major concern worldwide. There is an urgent need to identify bacterial factors promoting survival and persistence of these organisms in the nosocomial environment. Here, we describe the presence of a gene cluster, containing the Clp ATPase ClpK, within a collection of Danish ESBL-producing E. coli isolates. The cluster conferred thermoprotection upon the isolates, and thus might facilitate survival on medical devices exposed to semi-high temperatures in a hospital setting.


[Suspension arthroplasty of the thumb carpometacarpal joint in different views--a questionnaire].
Obviously there are different judgements of the results of suspension arthroplasties of the thumb carpometacarpal joint. Consequently, the three main groups involved in therapy were asked about this topic. 51 patients who had had a suspension arthroplasty on the thumb carpometacarpal joint in the period 1998 - 2006 were questioned. 12 of these patients were operated on both hands (63 operations). A questionnaire sent to 250 family doctors was compared with the results from the 51 patients. Furthermore there was an interview by telephone which included 25 practising specialists (orthopaedicians and surgeons). The results contrasted widely. Concerning the operation, 50 from 51 patients had a high subjective satisfaction. The family doctors had a different view. Nearly 80 % disregarded the success of the operation or they could not judge the chances. Only 50 % of all family doctors arranged for an X-ray or a consultation at a specialist when the patient had continuing problems. The results of the 25 specialists were also not uniform. 50 % of them, especially the hand surgeons, believe in the success of the primary osteoarthritis operation. The high barrier to operations for rhizarthrosis is based on a lack of information.


Comprehensive analysis of a pyroptosis-related gene signature of clinical and biological value in acute myeloid leukaemia.
Acutemyeloidleukaemia(AML) is an illness of varied origin and unpredictable prognosis. Pyroptosis, a novel class of gasdermin-mediated programmed cell death (PCD), serves a critical function in anti-leukemia. But, the correlation between pyroptosis-related genes (PRGs) and AML prognosis is undetermined. Here, we obtained the RNA profile and matched clinical information of AML patients from the TCGA and GEO databases. 6 PRGs were identified to be strongly related to AML prognosis via univariate COX analysis. Next, the LASSO regression analysis was used to develop a PRG signature for AML prognosis, which was then employed for the stratification of patients into a low- (LR) or high-risk (HR) group. Kaplan-Meier analysis revealed that the HR group, but not the LR group, had worse prognosis. In addition, ROC curve analysis revealed that our prognotic model had good predictive value. Functional enrichment analysis indicated that the immune status was remarkably different between the two risk groups. In vitro experiments demonstrated that pyroptosis serves an essential function in anti-leukemia treatment. In summary, our newly developed model has good predictive value and can offer guidance into the precise estimation of AML prognosis and targeting pyroptosis is a potential therapeutic alternative for AML.


Construction of a short form of the healthy eating behaviour inventory for the Japanese population.
The present study constructed a short form of the Healthy Eating Behavior Inventory suitable for use in the Japanese culture (HEBI-J) and confirmed the scale's preliminary reliability and validity. In Study 1, Japanese adults (N=75) completed a free-response questionnaire about healthy eating behaviours. Thematic analysis was used to identify three factors-balance, pattern, and restriction-associated with healthy eating behaviours and 12 items related to these healthy eating behaviours. In Study 2, Japanese office workers (N=784) completed two questionnaires regarding 12 items of healthy eating behaviours and the dietary stages of change. Confirmatory factor analysis demonstrated that the three-factor model was appropriate (GFI=.94, AGFI=.90, CFI=.90, RMSEA=.08, SRMR=.53). Reliability was confirmed by alpha coefficients and the Spearman-Brown formula. Concurrent validity was confirmed by the relationship between the healthy eating stages. This study demonstrated the preliminary reliability and validity of the HEBI-J.