[Comparison of two doses of intra-articular tranexamic acid on postoperative bleeding in total knee arthroplasty: a randomized clinical trial].
Tranexamic Acid (TXA), an antifibrinolytic that inhibits the fibrinolytic activity of plasmin is used to decrease perioperative blood loss and transfusion requirements in orthopedic surgery. The aim of our study was to compare postoperative bleeding in two intra-articular doses (1g and 2g) of tranexamic acid in adult patients undergoing unilateral total knee replacement. We conducted a single-operator, randomized, and controlled, double-blind study in two groups. The G1 group received 1g of intra-articular TXA and the G2 group 2g of intra-articular TXA. Both groups received 15mg.kg-1 IV before the surgical incision (TXA induction dose) and then 10mg.kg-1, orally, 6 and 12hours after the induction dose of TXA. The primary endpoint was bleeding measured by blood loss in postoperative drainage. Secondary outcomes were change in hemoglobin and hematocrit levels on the first and third postoperative days, and the need for transfusion during hospitalization. In total, 100 patients were randomized, and 100 were included in the analysis. Blood loss in postoperative drainage was similar in both groups (200±50 vs. 250±50mL, G1 and G2 groups respectively). Change in hematocrit and hemoglobin values (% of change) between preoperative and day 3 were not statically significant between groups G1 and G2 (18±5 vs. 21±4; 21±7 vs. 22±5 respectively). No patients received blood transfusion. Our study did not show superiority of 2g of intra-articular tranexamic acid compared to 1g. ClinicalTrials.gov Identifier NCT04085575.


Induction of ornithine decarboxylase activity in mouse tissues by phorbol ester is effectively blocked by methylglyoxal bis(butylamidinohydrazone).
Ornithine decarboxylase (ODC) was induced in the liver, lung and brain of the mouse injected intraperitoneally with 12-O-tetradecanoylphorbol 13-acetate (TPA), showing maximal enzyme activity four hours after the injection. The increase of ODC activity was due to the enhanced syntheses of mRNA and protein. The induction of ODC activity by TPA was specifically blocked by methylglyoxal bis(butylamidinohydrazone) (MGBB), a competitive inhibitor of ODC and S-adenosylmethionine decarboxylase, but not by the analog methylglyoxal bis(guanylhydrazone) (MGBG).


Pheochromocytoma, polycythemia, and venous thrombosis.
Polycythemia is rarely associated with pheochromocytoma. A patient with a 22-year history of malignant pheochromocytoma is presented in whom major complications developed as a result of long-standing polycythemia, apparently due to secretion of erythropoietin by the tumors. Despite attempts to reduce tumor burden by surgery, chemotherapy, and large doses of I-131-metaiodobenzylguanidine, polycythemia persisted. Extensive venous thrombosis developed requiring hospitalization and anticoagulation. Thus, polycythemia itself may be a cause of major morbidity in patients with pheochromocytoma, and prophylactic measures may be warranted. Review of the 130 patients with benign and malignant pheochromocytoma studied since the introduction of I-131-metaiodobenzylguanidine in 1980 revealed another six patients with hematocrits over 50 but only one had a hematocrit greater than 55 and required regular phlebotomy. In contrast, anemia (hematocrit less than 35) due to variety of causes was present in 18 cases.


Purification of a neuroprotective component of Parawixia bistriata spider venom that enhances glutamate uptake.
(1) In this study, we examined the effects of crude venom from the spider Parawixia bistriata on glutamate and GABA uptake into synaptosomes prepared from rat cerebral cortex. Addition of venom to cortical synaptosomes stimulated glutamate uptake and inhibited GABA uptake in a concentration-dependent manner. (2) The venom was fractionated using reverse-phase high-performance liquid chromatography on a preparative column. The fraction that retained glutamate uptake-stimulating activity was further purified on a reverse-phase analytical column followed by ion-exchange chromatography. (3) The active fraction, referred to as PbTx1.2.3, stimulated glutamate uptake in synaptosomes without changing the K(M) value, and did not affect GABA uptake. Additional experiments showed that the enhancement of glutamate uptake by PbTx1.2.3 occurs when ionotropic glutamate receptors or voltage-gated sodium and calcium channels are completely inhibited or when GABA receptors and potassium channels are activated, indicating that the compound may have a direct action on the transporters. (4) In an experimental model for glaucoma in which rat retinas are subjected to ischemia followed by reperfusion, PbTx1.2.3 protected neurons from excitotoxic death in both outer and inner nuclear layers, and ganglion cell layers. (5) This active spider venom component may serve as a basis for designing therapeutic drugs that increase glutamate clearance and limit neurodegeneration.


Thrombopoietin (TPO) regulates HIF-1alpha levels through generation of mitochondrial reactive oxygen species.
Hypoxia inducible factor (HIF)-1 is a master transcriptional regulator mediating the cellular adaptation to hypoxia. In addition, HIF-1 is also vital for the development of hematopoietic stem cells (HSCs). In a previous study we found that thrombopoietin (TPO), an important and non-redundant cytokine for HSC maintenance and expansion, induces HIF-1alpha expression in HSCs by enhancing the stability of HIF-1alpha under normoxic conditions. However, the molecular mechanisms of these effects are not yet fully understood. In this study, we explored the mechanisms and found that TPO-induced mitochondrial reactive oxygen species (ROS) played a crucial role in stabilization of HIF-1. Both ROS scavengers and inhibitors of mitochondrial electron transport completely blocked HIF-1alpha induction by TPO in UT-7/TPO cells and in primary immature mouse bone marrow cells. We also found that TPO-induced HIF-1alpha induction was tightly coupled with glucose metabolism. Inhibition of glucose transporter or glycolytic enzyme blocked HIF-1alpha elevation of TPO. These results indicate that TPO induces HIF-1alpha expression in a manner very similar to that of hypoxia.


Retracted: Simulation Analysis and Control of Multi-energy System for Fuel Cell Hybrid Electric Vehicle Based on Wavelet Transform.
[This retracts the article DOI: 10.1155/2022/3011307.].


[Update in continuous renal replacement techniques].
Acute renal failure affects 25% of patients hospitalized in intensive care units. Despite technological advances, the mortality of these patients is still high due to its associated complications. Continuous renal replacement techniques are one of the treatments for acute renal failure because they make it possible to treat the complications and decrease mortality. The nurse's knowledge and skills regarding these techniques will be decisive for the success of the therapy. Consequently, the nurse's experience and training are key components. The objective of this article is to update the knowledge on continuous renal replacement techniques. Keeping this in mind, a review has been made of the physical and chemical principles such as diffusion and convection, among others. A description of the different continuous renal replacement techniques, a presentation of the main vascular access, and a description of the nursing cares and complications related to techniques used have also been provided.


Effect of miR-451 on IVF/ICSI-ET outcome in patient with endometriosis and infertility.
This study aimed to explore the effect of miR-451 on IVF/ICSI-ET outcome in endometriosis patients with infertility. Eighty patients with endometriosis and infertility who came to our hospital for IVF/ICSI-ET from February 2018 to November 2019 were collected as the research participants, and 66 healthy women at the same time were selected as the control group. The miR-451 and MIF expression levels in serum, tissues and cell lines of patients with endometriosis and infertility were quantitatively detected by qRT-PCR. Correlation between miR-451 and endometriosis complicated with infertility was analyzed. The effect of miR-451 on IVF/ICSI-ET outcome in those patients was assessed. The miR-451 and MIF expression levels in endometriosis complicated with infertility tissues and cell lines were quantitatively detected by qRT-PCR. Compared with normal people, miR-451 was abnormally low in endometriosis complicated with infertility tissues and cell lines (P<0.001), while MIF was abnormally high (P<0.001), and the miR-451 expression was dramatically down-regulated and the MIF expression was markedly up-regulated in serum of endometriosis patients complicated with infertility. ROC analysis identified that the area under the miR-451 curve (AUC=0.9078) was >0.8, and the AUC (0.8606) of MIF was >0.8. Correlation analysis showed that the expression of miR-451 and MIF was negatively correlated in endometriosis complicated with infertility. According to miR-451 expression in endometriotic lesions, the subjects were divided into the miR-451 high expression group and miR-451 low expression group, with 40 cases in each group. The pregnancy rate after IVF/ICSI-ET in patients with endometriosis and infertility with high expression of miR-451 was higher than that in those with low expression (P>0.05). The incidence of complications during pregnancy after IVF/ICSI-ET in patients with endometriosis and infertility with high expression of miR-451 was lower than that in those with low expression (P>0.05). The pregnancy outcome after IVF/ICSI-ET in the miR-451 high expression group was better than that in the miR-451 low expression group (P<0.05). miR-451 was down-regulated in endometriosis patients complicated with infertility, and low expression of miR-451 after IVF/ICSI-ET indicated a poor outcome.


Hair analysis, a reliable and non-invasive method to evaluate the contamination by clenbuterol.
The illegal use of clenbuterol has been an increasingly serious issue in today's livestock products industry. It becomes an important project to develop a reliable approach to detect its content in food animals. A simple and sensitive LC-MS/MS method was developed to detect clenbuterol residue in hair, with the low limit of quantitation (LLOQ) about 0.5ng/g. Hogs fed with 340µg/day of clenbuterol for 2 weeks were found a high clenbuterol residue in their hair approximately at 1-2 months after withdrawal. There remained 3.31ng/g clenbuterol in hog hair approximately 5 months after the last administration, focused on the tip of the hair (mainly in hogs with dark hair). An extensive contamination was observed in twenty investigated market hogs whose dark hair obviously had a higher clenbuterol residue than the light ones (p=0.017, t test). Volunteers (60.3 percent) from Xuhui district (Shanghai) were found to have a detectable amount of clenbuterol in their hair (>0.5ng/g). In conclusion, hair residue detection is a reliable method to evaluate the clenbuterol contamination in animals and humans. Meat supply in the Xuhui district might have serious potential safety risks which should be further investigated and discussed to determine the safety range of clenbuterol residue.


Identification of functional methylated CpG loci in PD-L1 promoter as the novel epigenetic biomarkers for primary gastric cancer.
Gastric cancer (GC) is considered one of the most lethal malignancies worldwide due to poor prognosis. Aberrant methylation has been demonstrated to be involved in PD-L1 dysregulated expression in human cancers and possesses a great value as a diagnostic biomarker. Given that, in this study, we investigated the methylation status of PD-L1 as a promising biomarker in primary gastric tumors and identified functional CpG loci undergoing aberrant methylation through tumorigenesis of GC. PD-L1 methylation was initially evaluated in-silico using TCGA-STAD dataset. Pearson's correlation analysis was further employed to identify the most significant functional methylated CpG loci of PD-L1 gene in TCGA-STAD patient cohort. Methylation status and its correlation with PD-L1 expression were also validated using q-MSP and qRT-PCR in a set of internal samples, including 25 paired primary gastric tumors and adjacent normal tissues. The obtained results from TCGA-STAD showed that PD-L1 is significantly hypermethylated through gastric tumorigenesis, mostly in two CpG loci overlapping with cg19724470 and cg15837913 probes. Besides, PD-L1 DNA methylation was negatively correlated with PD-L1 expression in tumor samples. Furthermore, hypermethylation of cg19724470 and cg15837913 regions was validated in primary gastric tumors compared to adjacent normal samples. Also, ROC curve analysis illustrated the high diagnostic value of PD-L1 methylation for early detection of GC (AUC = 0.8110). In conclusion, the findings of this study suggested that PD-L1 expression is regulated by methylation in functional CpG loci and its methylation could be considered as a valuable diagnostic target for GC.


Expression of melanocortin-1 receptor in normal, malformed and neoplastic skin glands and hair follicles.
Melanocortin receptors (MC-Rs) are G-protein coupled receptors that mediate pleiotropic actions of melanocyte-stimulating hormones and adrenocorticotropin. There is increasing evidence that one of the five so far identified melanocortin receptors, i.e. melanocortin-1 receptor (MC-1R), has a more ubiquitous distribution in the skin than originally expected. In the present study, the expression of MC-1R in normal skin glands and hair follicles, various malformations and neoplasms with adnexal differentiation is described. Using an anti-MC-1R antibody directed against the amino acids 2-18 of the human MC-1R, specimens of normal healthy skin (n = 10) as well as hamartomas, cysts, hyperplasias, and benign or malignant neoplasms with eccrine, apocrine, sebaceous gland, and hair follicle differentiation (n = 98) were immunostained. MC-1R expression was widely preserved in various adnexal malformations and neoplasms as compared with normal skin and did not show major differences with regard to maturation of the neoplasms. The majority of adnexal epithelia showed an intracytoplasmically granular staining and, to a lesser extent, an intercellular staining pattern. Immunoelectron microscopical investigations revealed expression of MC-1R both along the cell surface and intracytoplasmically within tubular endosomes, the latter suggesting internalisation of the receptor. In conclusion, preserved MC-1R expression in adnexal epithelia suggests a functional role of proopiomelanocortin (POMC) in various malformations and neoplasms of the skin.


Ocular mycosis at a referral center in Saudi Arabia: A 20-year study.
To review the clinical experience of fungal keratitis cases at King Khaled Eye Specialist Hospital (KKESH) in Riyadh, Saudi Arabia. Retrospective observational review and analysis of 124 patient charts with confirmed diagnosis of fungal keratitis between 1984 and 2004. One hundred and twenty four eyes of 124 patients had proven fungal infection; 101 eyes had fungal keratitis and 23 eyes had fungal endophthalmitis complicating keratitis. Estimated proportion of fungal keratitis and endophthalmitis was 10.3%. Mean age was 55 years with male predominance (79.0%). Commonly associated factors were previous intraocular surgery (38.7%) and trauma (20.9%). Major risk factor for progressing to endophthalmitis was previous intraocular surgery (65.2%), p < 0.001. Initial laboratory results were fungal positive only in 30.6% (p < 0.001). Commonest organisms isolated were Aspergillus spp. (29.8%) followed by Trichophyton sp. (16.1%), then Candida and Fusarium sp. Comparison of both phases of the study showed improvement in the rate of successfully treated cases from 34.6% to 58.3%, and a decline in cases progressing to endophthalmitis from 25.0% to 13.9%. Therapeutic penetrating keratoplasty increased from 26.9% to 73.6% (p < 0.001). Thirteen eyes required enucleation or evisceration. In contrast to other studies on fungal keratitis, Aspergillusspp. and Trichophyton sp. were the most commonly isolated fungal pathogens; the former carries the worst prognosis. Risk factors included previous intraocular surgery and trauma. Poor outcome was associated with Aspergillus spp., delayed presentation, previous intraocular surgery and late surgical intervention. This study recommends early surgical intervention to improve the outcome.


Body condition is associated with a shorter breeding season and reduced ovulation rate in subtropical goats.
To determine the effect of body condition on the characteristics of the breeding season in female goats from subtropical Mexico, does in either greater- or lesser-body condition (n=20/group) were monitored for the expression of estrous behavior, ovulation and ovulation rate between June and April. The commencement of estrus and ovulation occurred earlier (P<0.05) in does in greater than lesser body condition. The cessation of estrus and ovulation was later (P<0.05) in female goats with greater than those with lesser body condition. Does in greater body condition had more (P<0.001) normal estrous cycles than those in lesser body condition. As consequence, does from the greater body condition group had fewer short (P<0.001) or long (P<0.05) estrous cycles than those of in the lesser body condition group. The ovulation rate was greater (P<0.01) in the greater (1.9+/-0.1) than lesser body condition does (1.6+/-0.1). In conclusion, female goats in lesser body condition have a shorter breeding season, more abnormal estrous cycles, and fewer ovulations than does in greater body condition.


Comparison of posttranslational protein modification by amino acid addition after crush injury to sciatic and optic nerves of rats.
Posttranslational protein modifications by the addition of amino acids are reactions which occur in intact sciatic and optic nerves of rats. The nerves differ, however, in that 2 h after crush injury these reactions are activated in sciatic but not in optic nerves. As sciatic nerves will eventually regenerate, whereas optic nerves will not, we have proposed that the activation of these reactions is correlated with the ability of a nerve to regenerate. The current experiments examined the posttranslational addition of amino acids to proteins at times greater than 2 h after nerve crush, during sciatic nerve regeneration and optic nerve degeneration. We also examined the optic nerve for morphologic correlates to changes in protein modification and partially characterized the proteins modified by [3H]Lys in the regenerating sciatic nerve using two-dimensional sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE). In a segment of sciatic nerve taken from a region just proximal to the site of crush, protein modification by covalent addition of [3H]Arg, [3H]Lys and [3H]Leu increased during both posttraumatic (2 h postcrush) and regenerative (6 days and 14 days postcrush) stages. Two-dimensional PAGE of [3H]Lys modified sciatic nerve proteins 6 days after crush injury showed labeling of proteins having molecular masses in the 18,000- to 20,000-, 30,000- to 40,000-, and 80,000- to 100,000-Da ranges, with neutral or basic isoelectric points (pI 7.1 to 8.0). In the retinal portion of the crushed optic nerve, incorporation of the same amino acids was unchanged or depressed to 21 days postcrush, except at 6 days postcrush when the incorporation of all three amino acids into proteins was increased threefold. These increases correlated with the appearance of terminal end bulbs in the portion of nerve analyzed. Histological examination of each nerve 2 h postcrush showed marked edema in the optic but not the sciatic nerve, a condition which may be related to the ability of sciatic and inability of optic nerves to activate protein modification reactions.


Physiotherapy management of nociplastic pain: A Delphi study of Italian specialists.
Nociplastic pain due to central sensitization (CS) is common in people suffering from chronic pain, but no clinical practice guideline is available in rehabilitative settings for patients' management. The aim of this study is to achieve expert consensus on physiotherapy competencies in the management of people with nociplastic pain and suspected CS mechanisms. A web-based Delphi process was employed. Experts in the rehabilitation field were recruited following pre-defined eligibility criteria. Following completion of three Delphi rounds, the final list of competencies was generated. In all, 23 participants were recruited. They all completed Round 1 (23/23, 100%), twenty Round 2 and Round 3 (20/23, 87%). Following Round 1, seven areas were identified by the panel as crucial for CS physiotherapy management; 19 competencies out of 40 reached the consensus between experts, and nine additional competencies were added to Round 2 following literary review. Round 2 identified the agreement for all the 29 competencies. During Round 3, all the experts confirmed the final list generated through the consensus process. An agreement between experts was found for the final list of competencies that a physiotherapist should implement every time it approaches people with suspected CS mechanisms. Further research is needed to support the clinical utility of our findings and their applicability in daily practice.


Seroprevalence of respiratory syncytial virus in free-ranging bighorn sheep.
Sera from 187 (42%) of 447 free-ranging bighorn sheep (Ovis canadensis) in 9 western states had antibody titers to respiratory syncytial virus (RSV). Titers were from 1:5 to 1:640, as determined by the standard virus neutralization test. Results of the survey indicated that RSV is widespread in free-ranging bighorn sheep populations and that RSV may be an important factor in the bighorn sheep pneumonia-complex.


Sexual functioning, sexual enjoyment, and body image in Norwegian breast cancer survivors: a 12-year longitudinal follow-up study and comparison with the general female population.
Given the scarcity of evidence concerning the long-term sexual health of breast cancer (BC) survivors (BC-Pop), we aimed to assess how BC treatments affect short- and long-term sexual functioning, sexual enjoyment, and body image, and compare with aged-matched women in the Norwegian general population (F-GenPop). The 349 patients in BC-Pop treated at Trondheim University Hospital in 2007-2014, were assessed in clinical controls at the hospital; before starting radiotherapy (T1, baseline), immediately after ending radiotherapy (T2), and after 3, 6, and 12 months (T3-T5), and at a long-term follow-up 7-12 years after baseline (T6). Meanwhile, F-GenPop included 2254 age-matched women in the Norwegian general population. The impact of BC treatment on sexual functioning was examined using a Linear Mixed Model. Sexual functioning, sexual enjoyment, and body image were assessed with the EORTC's QLQ-BR23 scales and compared between the populations in the four age groups (30-49, 50-59, 60-69, and 70+ years) using means with 95% confidence intervals and Student t-test. Linear regression, adjusted for age and comorbidity was applied to estimate individual scores. BC survivors treated with mastectomy had overall lower sexual functioning than patients who had received breast-conserving surgery (p = 0.017). Although BC survivors treated with chemotherapy had lower sexual functioning than those treated without chemotherapy at T1-T5 (p = 0.044), both groups showed the same level of functioning at T6. BC-Pop exhibited significantly poorer sexual functioning (p < 0.001), lower sexual enjoyment (p < 0.05), and better body image (p < 0.001) than F-GenPop in all age groups. The impact of specific BC treatments on sexual functioning was modest; only mastectomy had a persistent negative influence. Nevertheless, all age groups in BC-Pop displayed significantly poorer sexual functioning than F-GenPop at both 12 months and up to 12 years after treatment.


The solution NMR structure of Antheraea polyphemus PBP provides new insight into pheromone recognition by pheromone-binding proteins.
Pheromone-binding proteins (PBPs) located in the antennae of male moth species play an important role in olfaction. They are carrier proteins, believed to transport volatile hydrophobic pheromone molecules across the aqueous sensillar lymph to the membrane-bound G protein-coupled olfactory receptor proteins. The roles of PBPs in molecular recognition and the mechanisms of pheromone binding and release are poorly understood. Here, we report the NMR structure of a PBP from the giant silk moth Antheraea polyphemus. This is the first structure of a PBP with specific acetate-binding function in vivo. The protein consists of nine alpha-helices: alpha1a (residues 2-5), alpha1b (8-12), alpha1c (16-23), alpha2 (27-34), alpha3a (46-52), alpha3b (54-59), alpha4 (70-79), alpha5 (84-100) and alpha6 (107-125), held together by three disulfide bridges: 19-54, 50-108 and 97-117. A large hydrophobic cavity is located inside the protein, lined with side-chains from all nine helices. The acetate-binding site is located at the narrow end of the cavity formed by the helices alpha3b and alpha4. The pheromone can enter this cavity through an opening between the helix alpha1a, the C-terminal end of the helix alpha6, and the loop between alpha2 and alpha3a. We suggest that Trp37 may play an important role in the initial interaction with the ligand. Our analysis also shows that Asn53 plays the key role in recognition of acetate pheromones specifically, while Phe12, Phe36, Trp37, Phe76, and Phe118 are responsible for non-specific binding, and Leu8 and Ser9 may play a role in ligand chain length recognition.


["Assure" program, to improve the management of emergency situations in Ehpad].
The Assure approach (improvement of emergency care) is a project designed to improve emergency care for the 63,000 residents of homes for the dependent elderly (Ehpad) in the Ile-de-France region. By reinforcing the skills of caregivers in emergency situations and facilitating collaboration between care providers, the Assure approach is mobilizing, alongside all the Ehpad in the Ile-de-France region over a period of two years, the emergency medical assistance services, emergency physicians, mobile geriatric teams and nursing and care assistant training institutes.


Inhibition of myogenesis by okadaic acid, an inhibitor of protein phosphatases, 1 and 2A, correlates with the induction of AP1.
Recently, we demonstrated that okadaic acid, an inhibitor of protein phosphatases 1 and 2A, inhibits myogenesis by extinguishing the expression of MyoD1 and inducing the expression of Id. Since it has been reported that transformation by c-fos also inhibits myogenesis through inhibition of MyoD1 expression, we examined the effects of okadaic acid on the activation of the c-fos and jun family of proto-oncogenes in an attempt to understand the mechanism by which okadaic acid inhibits the myogenic differentiation. Treatment of C2C12 cells in growth medium with okadaic acid increased expression of the mRNAs for the c-fos family continuously and for the jun family to a lesser extent. In contrast, in differentiation medium, the induction of c-fos, c-jun, and fos B mRNAs by okadaic acid was transient, whereas fra-1, jun D, and jun B mRNAs were induced continuously, suggesting that okadaic acid regulates the expression of the c-fos and jun family through complex regulatory mechanisms depending on the state of differentiation of the cells. Transfection of c-jun and c-fos promoter-chloramphenicol acetyltransferase constructs demonstrated that the effects of okadaic acid on the induction of c-fos and c-jun are mediated through the activation of promoter elements. These results suggest that some of the targets of protein phosphatases 1 and 2A may include transcription factors capable of forming AP1 complexes and that these factors may play an important role during myogenic differentiation.


Tissue engineering technology and its possible applications in oral and maxillofacial surgery.
Tissue engineering is a rapidly advancing discipline that combines the attributes of biochemical and biomaterial engineering with cell transplantation to create bio-artificial tissues and organs. For the oral and maxillofacial surgeon, the reconstruction of maxillofacial defects in hard and soft tissues is an ongoing challenge. While autologous grafts and vascularised free flaps are the current gold standard, they are not without complications at both the donor and reconstructed sites. Tissue engineering, which aims to create tissue-matched, prefabricated, prevascularised bony or soft tissue composite grafts, or both, therefore has the potential to revolutionise practice in maxillofacial surgery. We review the technology of tissue engineering and its current and future applications within the specialty, and discuss contemporary obstacles yet to be overcome.


The effects of modified atmospheres on the growth of psychrotrophic pseudomonads on a surface in a model system.
Atmospheres containing concentrations of CO2 as low as 20% (balance nitrogen) inhibited the growth of Pseudomonas fluorescens and Pseudomonas putida on the surface of buffered Brain Heart Infusion agar plates, pH 6.8, incubated at 5 or 15 degrees C in flexible packages. The modified atmospheres decreased the growth rates and reduced the populations attained at the end of the exponential phase of growth, but had no substantial effect on the lag phase. P. fluorescens was less tolerant of CO2 than P. putida. The inhibitory effect of CO2 increased with its concentration and inhibition was greater at 5 than at 15 degrees C. Growth occurred in packages flushed with 20, 40 and 100% CO2 and 100% N2 at 15 degrees C and 20 and 40% CO2 and 100% N2 at 5 degrees C. The residual O2 concentration in the packages after flushing was 0.2-0.5%. Storage of pseudomonads in CO2 under conditions that prevented growth (e.g., 100% CO2, 5 degrees C) did not cause substantial loss of viability. There was no detectable residual effect of CO2. If cultures were incubated in air after storage for up to 70 days in CO2-containing atmospheres which prevented growth, the subsequent growth curve did not differ noticeably from that observed when plates were incubated in air immediately after inoculation. When cultures in the exponential or stationary phases of growth in modified atmospheres were transferred to air, growth rates increased quickly to rates similar to those observed in air and the final populations observed in air were attained. A reduction in the pH of the medium to 5.5 substantially increased the inhibitory effect of CO2. At 5 degrees C and pH 5.5, substantial growth of P. fluorescens was not observed in any of the CO2 concentrations tested, nor in 40 or 100% CO2 for P. putida.


Lanthanide Metal-Organic Framework Nanoprobes for the In Vitro Detection of Cardiac Disease Markers.
Acute myocardial infarction (AMI) is one of the leading causes of death around the world. An early and accurate diagnosis of AMI is critical to reduce the mortality rate. As an important cardiac biomarker, creatine kinase (CK) has been used in the clinical diagnosis of AMI. However, it still remains a great challenge to realize highly sensitive and selective CK detection in blood specimens. Herein, we have developed an ultrasensitive platform for the detection of CK activity based on time-resolved (TR) luminescent lanthanide metal-organic framework nanoprobes (Eu-QPTCA). Benefiting from the intense emission of lanthanide ions sensitized by the organic ligands and the eliminated short-lived autofluorescence by the TR technique, these nanoprobes enabled the homogeneous detection of CK activity with a limit of detection down to 1.0 U/L, which is about 1 order of magnitude improvement relative to that of the traditional methods. In addition, the Eu-QPTCA nanoprobes showed superior selectivity and reliability toward the practical detection of CK activity in human serum, indicating the great significance of our method in the early diagnosis of AMI. We envision that the proposed bioassay strategy can be extended to the detection of other phosphorylation enzymes, paving a way for promising applications in clinical diagnostics.


Studies in vitro on the inactivation of mitochondrial rat-liver aldehyde dehydrogenase by the alcohol-sensitizing compounds cyanamide, 1-aminocyclopropanol and disulfiram.
The inhibition of the low-Km, rat-liver mitochondrial aldehyde dehydrogenase (ALDH) by the alcohol-sensitizing agents cyanamide, 1-aminocyclopropanol (ACP) and disulfiram was studied in vitro. All three compounds caused a progressive decline in the enzyme activity. Restoration of activity could not be achieved by gel-filtration, dilution or by the addition of excess thiol. High concentrations of acetaldehyde partly restored the activity of the cyanamide-inactivated enzyme but had no effects on the disulfiram- or ACP-inactivated enzyme. In the presence of saturating concentrations of the coenzyme (NAD+), the inactivation process followed first-order kinetics at fixed concentrations of the inhibitors. Plots of the apparent first-order rate constants against inhibitor concentration were curved, suggesting the formation of saturable, reversible holoenzyme-inhibitor complexes prior to the covalent reactions. In the absence of NAD+, the rate of inactivation by disulfiram was biphasic and considerably higher than that in the presence of NAD+. In contrast, no inactivation was obtained with cyanamide in the absence of NAD+. Likewise, the presence of NAD+ greatly promoted the inactivation by ACP. The esterase activity of the enzyme was also affected by the inhibitors, although to a lesser extent than was the dehydrogenase activity. The results obtained suggest that all three inhibitors inactivate the enzyme through covalent reactions with the thiol groups at the active site. It is proposed that binding of NAD+ limits access of disulfiram to the thiols at the active site but provides a situation that favours an electrophilic attack of cyanamide and ACP on the thiol groups.


Reliability of two novel methods, Alamar and E test, for detection of methicillin-resistant Staphylococcus aureus.
E test strips (AB Biodisk, Culver City, Calif.) tested on 2% NaCl-supplemented Mueller-Hinton agar and Alamar panels (Alamar, Sacramento, Calif.) correctly characterized 127 of 127 methicillin-resistant Staphylococcus aureus isolates and 100 of 100 methicillin-susceptible S. aureus isolates. These overnight antimicrobial susceptibility test systems reliably detect this clinically important resistance.


Radical treatment of synchronous oligometastatic non-small cell lung carcinoma (NSCLC): patient outcomes and prognostic factors.
Metastatic non-small cell lung carcinoma (NSCLC) generally carries a poor prognosis, and systemic therapy is the mainstay of treatment. However, extended survival has been reported in patients presenting with a limited number of metastases, termed oligometastatic disease. We retrospectively reviewed the outcomes of such patients treated at two centers. From September 1999-July 2012, a total of 61 patients with 1-3 synchronous metastases, who were treated with radical intent to all sites of disease, were identified from records of two cancer centers. Treatment was considered radical if it involved surgical resection and/or delivery of radiation doses ≥13 × 3 Gy. Besides the primary tumor, 50 patients had a solitary metastasis, 9 had two metastases, and 2 had three metastases. Locations of metastases included the brain (n = 36), bone (n = 11), adrenal (n = 4), contralateral lung (n = 4), extra-thoracic lymph nodes (n = 4), skin (n = 2) and colon (n = 1). Only one patient had metastases in two different organs. Median follow-up was 26.1 months (m), median overall survival (OS) was 13.5m, median progression free survival (PFS) was 6.6m and median survival after first progression (SAFP) was 8.3m. The 1- and 2-year OS were, 54% and 38%, respectively. Significant predictors of improved OS were: smaller radiotherapy planning target volume (PTV) (p = 0.004) and surgery for the primary lung tumor (p < 0.001). Factors associated with improved SAFP included surgery for the primary lung tumor, presence of brain metastases, and absence of bone metastases. No significant differences in outcomes were observed between the two centers. Radical treatment of selected NSCLC patients presenting with 1-3 synchronous metastases can result in favorable 2-year survivals. Favorable outcomes were associated with intra-thoracic disease status: patients with small radiotherapy treatment volumes or resected disease had the best OS. Future prospective clinical trials, ideally randomized, should evaluate radical treatment strategies in such patients.


Targeting nicotinamide N-methyltransferase overcomes resistance to EGFR-TKI in non-small cell lung cancer cells.
Activating mutations of epidermal growth factor receptor (EGFR) contributes to the progression of non-small cell lung cancer (NSCLC). EGFR tyrosine kinase inhibitor (TKI)-targeted therapy has become the standard treatment for NSCLC patients with EGFR-mutations. However, acquired resistance to these agents remains a major obstacle for managing NSCLC. Here, we investigated a novel strategy to overcome EGFR TKI resistance by targeting the nicotinamide N-methyltransferase (NNMT). Using iTRAQ-based quantitative proteomics analysis, we identified that NNMT was significantly increased in EGFR-TKI-resistant NSCLC cells. Moreover, we found that NNMT expression was increased in EGFR-TKI-resistant NSCLC tissue samples, and higher levels were correlated with shorter progression-free survival in EGFR-TKI-treated NSCLC patients. Knockdown of NNMT rendered EGFR-TKI-resistant cells more sensitive to EGFR-TKI, whereas overexpression of NNMT in EGFR-TKI-sensitive cells resulted in EGFR-TKI resistance. Mechanically, upregulation of NNMT increased c-myc expression via SIRT1-mediated c-myc deacetylation, which in turn promoted glycolysis and EGFR-TKI resistance. Furthermore, we demonstrated that the combination of NNMT inhibitor and EGFR-TKI strikingly suppressed the growth of EGFR-TKI-resistant NSCLC cells both in vitro and in vivo. In conclusion, our research indicated that NNMT overexpression is important for acquired resistance to EGFR-TKI and that targeting NNMT might be a potential therapeutic strategy to overcome resistance to EGFR TKI.


Continuous epidural infusion of 0.05% bupivacaine plus hydromorphone for labor analgesia: an observational assessment in 1830 parturients.
We evaluated a continuous epidural infusion containing bupivacaine 0.05% plus the opioid hydromorphone in 1830 women requesting pain relief during labor and delivery. The infusion provided effective analgesia with minimal adverse events for patients differing in parity and at varying stages of labor. Pain relief was maintained in most patients without the need for epidural reinforcement with more concentrated doses of local anesthetic.


scBPGRN: Integrating single-cell multi-omics data to construct gene regulatory networks based on BP neural network.
The deterioration and metastasis of cancer involve various aspects of genomic changes, including genomic DNA changes, epigenetic modifications, gene expression, and other complex interactions. Therefore, integrating single-cell multi-omics data to construct gene regulatory networks containing more omics information is of great significance for understanding the pathogenesis of cancer. In this article, an algorithm integrating single-cell RNA sequencing data and DNA methylation data to construct a gene regulatory network based on the back-propagation (BP) neural network (scBPGRN) is proposed. This algorithm uses biweight extreme correlation coefficients to measure the correlation between factors and uses neural networks to calculate generalized weights to construct gene regulation networks. Finally, the node strength is calculated to identify the genes associated with cancer. We apply the scBPGRN algorithm to hepatocellular carcinoma (HCC) data. We construct a regulatory network and identify top-ranked genes, such as MYCBP, KLHL35, PRKCZ, and SERPINA6, as the key HCC-related genes. We analyze the top 100 genes, and the HCC-related genes are concentrated in the top 20. In addition, the single cell data is found to consist of two subpopulations. We also apply scBPGRN to two subpopulations. We analyze the top 50 genes in them, and the HCC-related genes are concentrated in the top 20. The consequences of functional enrichment analysis indicate that the gene regulatory network we have constructed is valid. Our results have been verified in several pieces of literature. This study provides a reference for the integration of single-cell multi-omics data to construct gene regulatory networks.


Long-term outcome after drug-eluting stent implantation in comparison with bare metal stents: a single centre experience.
The aim of our study was to evaluate the effect of drug-eluting stents (DES) compared with bare-metal stents (BMS) on all-cause mortality and target vessel revascularization (TVR) in a "real-world" clinical setting. One thousand four hundred and ninety consecutive patients, who underwent PCI, were included in a prospective registry from January 2003 until December 2006. Patients were divided retrospectively into two groups: those who received a DES and those who received a BMS. The primary combined endpoint all-cause mortality or TVR and the individual endpoints death and TVR were evaluated during a mean follow-up period of 24.56 ± 12.5 months (range 6-52 months). In total 1,033 patients (69.3%) received BMS, while 457 patients (30.7%) received DES. With respect to clinical characteristics, significant differences between groups were found for age, hyperlipidemia, diabetes mellitus, heart failure, previous cerebral insult and presence of acute coronary syndrome (ACS) during intervention. Propensity score analysis was performed in attempt to eliminate this selection bias. With respect to the combined endpoint all-cause death or TVR, 12.9% of patients with DES and 21.3% with BMS (p = 0.015) had an event during the follow-up. 12.3% of patients with BMS but only 5.7% with DES died during the follow-up (p = 0.025). Thirty-three patients (7.2%) of the DES group and 99 patients (9.6%) of the BMS group (p = 0.1) needed TVR. Our results obtained in a "real-world" clinical setting exhibit a clinical long-term benefit for DES compared with BMS and underline the safety and efficacy of DES over BMS.


Terminology for training of endoscopic submucosal dissection.
Like many other advanced endoscopic skills, to master the skill of endoscopic submucosal dissection (ESD) requires training for a novice. The general medical terminology should be used similarly even in case of ESD training. However, it is not common for everyone to recall the same meaning from one medical term. Therefore, it is necessary to unify the meaning of medical terms and review their usage in a meeting to achieve a consensus. For this purpose, terms used in the upper gastrointestinal session, Endoscopic Forum Japan 2011, entitled 'Towards further penetration of ESD techniques - what is the role of Japanese ESD experts?', were determined beforehand as shown. Additionally, the present educational approach of ESD in Japan is simply outlined in this article.


Microfluidic flow focusing: drop size and scaling in pressure versus flow-rate-driven pumping.
We experimentally study the production of micrometer-sized droplets using microfluidic technology and a flow-focusing geometry. Two distinct methods of flow control are compared: (i) control of the flow rates of the two phases and (ii) control of the inlet pressures of the two phases. In each type of experiment, the drop size l, velocity U and production frequency f are measured and compared as either functions of the flow-rate ratio or the inlet pressure ratio. The minimum drop size in each experiment is on the order of the flow focusing contraction width a. The variation in drop size as the flow control parameters are varied is significantly different between the flow-rate and inlet pressure controlled experiments.


Nitric oxide of human colorectal adenocarcinoma cell lines promotes tumour cell invasion.
The present study investigates the role of nitric oxide and the involvement of nitric oxide synthase II isoform on the invasion of human colorectal adenocarcinoma cell lines HRT-18 and HT-29. HRT-18 cells, which constitutively express nitric oxide synthase II mRNA were three-fold more invasive in a Matrigel invasion assay than nitric oxide synthase II mRNA negative HT-29 cells. Treatment of HT-29 cells with the nitric oxide donor Deta NONOate (50 nM) as well as induction of nitric oxide synthase II mRNA and production of endogenous nitric oxide by inflammatory cytokines (IFN-gamma and IL-1alpha) increased the invasiveness of HT-29 cells by approximately 40% and 75%, respectively. In HT-29 cells nitric oxide synthase II mRNA was also induced in co-culture with human monocytes. The invasiveness of HRT-18 cells and stimulated HT-29 cells was partly inhibited by the nitric oxide synthase II inhibitor 1400 W. These results show that nitric oxide increases the invasion of human colorectal adenocarcinoma cell lines HRT-18 and HT-29, and the involvement of nitric oxide synthase II isoform in tumour cell invasion. Therefore, the production of nitric oxide and secretion of pro-inflammatory cytokines by tumour-associated macrophages, which in turn induce nitric oxide synthase II isoform in tumour cells, promotes tumour cell invasiveness.


The differing responses of four muscle types to dexamethasone treatment in the rat.
The glucocorticoid dexamethasone dramatically altered growth patterns in four muscle types, inducing atrophy of smooth and fast-twitch skeletal muscle, suppressing protein accumulation in slow-twitch muscle and enhancing growth in the heart. These differing responses were explained by steroid-induced changes in RNA content, protein synthesis and protein breakdown.


Comparative chemical and immunological characterization of five lipolytic enzymes (carboxylesterases) from rat liver microsomes.
The chemical and immunological properties of five closely related microsomal serine hydrolases (carboxylesterases) from rat liver have been compared to evaluate whether they are variants of a single protein or independent proteins. These enzymes represent medium-chain-length acylcarnitine hydrolase, palmitoyl carnitine hydrolase, medium-chain-length monoglyceride hydrolase, and two long-chain monoglyceride hydrolases. All enzymes have similar subunit Mr's (58,000-61,000) and bear one active site per protein subunit, as could be shown by active sites with radioactive bis(4-nitrophenyl)phosphate, and have subsequently been cleft by proteases or by BrCN. The patterns of radioactive peptides obtained after electrophoresis or thin-layer chromatography indicated that the two long chain monoglyceride hydrolases were closely related, while all other hydrolases differed from these and from each other. The two long-chain monoglyceride hydrolases also had identical N- and C-termini that differed from those of the other hydrolases. All hydrolases contain low amounts of hexoses. It is concluded that the hydrolases investigated represent four independent enzymes with differing amino acid sequences. Three of the four hydrolases were microheterogenous. These results were confirmed with an immunological study using rabbit antisera against three of the hydrolases. Heparin-releasable liver lipase was not cross-reactive with the lipolytic enzymes investigated here.


The significance of changes in high mobility group-1 protein mRNA expression in rats after thermal injury.
There has been a widespread impression that tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) mediate the toxicity of high doses of lipopolysaccharide (LPS, endotoxin) and are key factors in septic shock. However, the clinical efficacy of treatment with antagonists of TNF-alpha and IL-1beta is still controversial, suggesting that mediators other than TNF-alpha and IL-1beta might contribute causally to endotoxin-induced death. Recent studies implicated high mobility group-1 (HMG-1) protein as a late mediator of endotoxin lethality in mice. However, the role of HMG-1 in mediating multiple organ damage-associating trauma has not been studied. This study was designed to investigate changes in HMG-1 gene expression in vital organs, and its potential role in mediating multiple organ damage following major burns. Wistar rats were subjected to a 35 percent full-thickness thermal injury, and randomly divided into three groups as follows: normal controls (n = 7), thermal injury (n = 24), and recombinant bactericidal/permeability-increasing protein (rBPI21) treatment (n = 12). Tissue samples from liver and lungs were collected to measure tissue endotoxin levels and HMG-1 mRNA expression. In addition, blood samples were obtained for measurement of organ function parameters. Our data demonstrated a significant increase in HMG-1 gene expression in tissues at 24 h postburn, which remained markedly elevated up to 72 h after thermal injury (P< 0.05-0.01). Treatment with rBPI21 could significantly decrease tissue HMG-1 mRNA expression in the liver and lung (P < 0.01). In addition, there were high positive correlations between hepatic HMG-1 mRNA and serum aminoleucine transferase (ALT) and aspartate aminotransferase (AST) levels, and also between pulmonary HMG-1 mRNA and myeloperoxidase activities (P < 0.05-0.01). Taken together, these findings indicate that thermal injury per se can markedly enhance HMG-1 gene expression in various organs. Up-regulation of HMG-1 expression may be involved in the pathogenesis of endogenous endotoxin-mediated multiple organ damage secondary to major burns.


Essentiality of fatty acid synthase in the 2D to anchorage-independent growth transition in transforming cells.
Upregulation of fatty acid synthase (FASN) is a common event in cancer, although its mechanistic and potential therapeutic roles are not completely understood. In this study, we establish a key role of FASN during transformation. FASN is required for eliciting the anaplerotic shift of the Krebs cycle observed in cancer cells. However, its main role is to consume acetyl-CoA, which unlocks isocitrate dehydrogenase (IDH)-dependent reductive carboxylation, producing the reductive power necessary to quench reactive oxygen species (ROS) originated during the switch from two-dimensional (2D) to three-dimensional (3D) growth (a necessary hallmark of cancer). Upregulation of FASN elicits the 2D-to-3D switch; however, FASN's synthetic product palmitate is dispensable for this process since cells satisfy their fatty acid requirements from the media. In vivo, genetic deletion or pharmacologic inhibition of FASN before oncogenic activation prevents tumor development and invasive growth. These results render FASN as a potential target for cancer prevention studies.


Lipoprotein apheresis and PCSK9 inhibitors for severe familial hypercholesterolaemia: Experience from Australia and New Zealand.
Severe familial hypercholesterolaemia (FH) causes premature disability and death due to atherosclerotic cardiovascular disease and is refractory to standard lipid-lowering therapies. Lipoprotein apheresis (LA) has long been a standard of care for patients with severe FH, but is invasive, expensive and time-consuming for patients and their caregivers. Newer drug therapies, including the proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors, may reduce the need for LA. We audited the records of 16 patients (eight homozygous, eight heterozygous) treated with LA in Australia and New Zealand, 14 of whom subsequently commenced PCSK9 inhibitor therapy. LA was performed by cascade filtration in all centres. LDL-cholesterol was acutely lowered by 69 ± 7% in patients with homozygous FH and by 72 ± 9% in those with heterozygous FH, representing time-averaged reductions of 36 ± 12% and 34 ± 5%, respectively. LA was well-tolerated, and patients reported comparable quality of life to population and disease-related norms. After commencement of PCSK9 inhibitors, four of seven patients with homozygous FH had meaningful biochemical responses, with a reduction in the frequency of LA permitted in one patient and complete cessation in another. Four of seven patients with heterozygous FH were able to be managed without LA after commencing PCSK9 inhibitors. While PCSK9 inhibitors have reduced the need for LA, some patients with severe FH continue to require LA, and will require it for the foreseeable future. However, emerging therapies, including angiopoetin-like 3 inhibitors, may further reduce the need for LA.


Continuous ventricular volume assessment for diagnosis and pacemaker control.
Stroke volume measurements made by a catheter-based, right ventricular, intracardiac impedance system at rest and during upright bicycle ergometry were found to compare favorably with measurements made by acetylene rebreathing (r = 0.96, n = 13) and by radionuclide ventriculography (r = 0.96, n = 13) in a 46-year-old male pacemaker-dependent subject. The impedance information was then used to control the ventricular pacing rate during bicycle exercise, utilizing an algorithm that attempts to maintain a constant stroke volume. This resulted in a 23% increase in cardiac output accompanied by a 70% decrease in stroke volume as compared to the values measured during the same exercise at 70 pulses per minute (ppm).


[Osteogenic tumors of bone].
Osteogenic tumors include malignant and benign tumors that produce tumor osteoid and/or bone tissue. Osteosarcoma is the most common malignant bone tumor, especially in children and young adults. The entities with their characteristic morphological features are described to enable the reader to come to a diagnosis and differential diagnosis on the basis of patient age, history and predominant location of the tumor. For this review we selectively used mainly large published patient cohorts. Our own and externally published data on widely accepted tumor criteria were also compared. Detection is the initial diagnostic step for an osseous lesion, and is determined by the sensitivity of the method applied. Plain X‑ray films in two planes and CT are the basics in the radiological toolkit for osteogenic tumors. For evaluation of local tumor extension and biopsy planning MRI or scintigraphy should be combined. MRI as a stand-alone diagnostic tool is insufficient. For malignant bone tumors staging should be performed, applying a variable combination of thoracic CT, MRI, scintigraphy, and positron emission tomography (PET). Osteosarcoma, along with Ewing sarcoma and chondrosarcoma, are the most common malignant bone tumors; all sub-entities are significantly rarer. Among benign bone tumors, osteoid osteomas have the highest incidence, presenting with typical pain, location, and age predilection. Diagnostics and treatment of malignant bone tumors should preferably be performed in specialized centers because of significant therapeutic implications for patients. In uncertain cases, a second opinion should always be obtained.


Purification and characterization of a thermo-acid/alkali stable xylanases from Aspergillus oryzae LC1 and its application in Xylo-oligosaccharides production from lignocellulosic agricultural wastes.
Xylooligosaccharides (XOS) from lignocellulosic biomass (LCB) have found widespread applications in food, feed, nutraceuticals and pharamecutical industries. Enzymatic degradation of LCB for generation of XOS have gained impetus in recent times In the present investigation an extracellular thermo-alkali stable xylanase from Aspergillus oryzae LC1 was purified by using PEG 8000/MgSO4 aqueous two-phase system and was capable of hydrolysing various agricultural residues into XOS system. Highest activity was observed using 11.3% (w/w) PEG 8000 and 22.5% (w/w) sulphate salt with maximum purification factor (13-fold), highest yield (86.8%) and partition coefficient (8.8%). The purification of the crude enzyme also resulted in decrement of β-xylosidase activity (29.8 U/mL to 0.6 U/mL). The molecular weight of enzyme was estimated ~35 kDa. The highest residual activity was obtained with birch wood xylan as substrate with Km and Vmax of 0.2 mg/mL and 172.2 μmol min-1 mg-1 respectively. The metal ions Fe2+, Ag2+, Mg2+, Mn+ and Co+ enhanced xylanase activity while EDTA, DMSO and SDS acted as inhibitor. The effect of Fe+2 was confirmed by the circular dichroism experiment. The partially purified enzyme was capable of generating XOS i.e. xylobiose (0.68 mg/g), xylotriose (2.47 mg/g) and xylotetraose (2.29 mg/g) by direct enzymatic hydrolysis of untreated sugarcane baggase, wheat straw and wheat bran respectively.


Making post-acute care assets viable: a system's approach to continuing care.
To build a strong continuing care network, leaders at Spectrum Health: Recruited industry veterans in post-acute care, Increased the visibility of the parent brand, Gained greater alignment throughout the system, Filled gaps in the health system's post-acute care portfolio.


Review of non-permeating cryoprotectants as supplements for vitrification of mammalian tissues.
Vitrification of mammalian tissues is important in the areas of human assisted reproduction, animal reproduction, and regenerative medicine. Non-permeating cryoprotectants (CPAs), particularly sucrose, are increasingly used in conjunction with permeating CPAs for vitrification of mammalian tissues. Combining non-permeating and permeating CPAs was found to further improve post-thaw viability and functionalities of vitrified mammalian tissues, showing the potential applications of such tissues in various clinical and veterinary settings. With the rising demand for the use of non-permeating CPAs in vitrification of mammalian tissues, there is a strong need for a timely and comprehensive review on the supplemental effects of non-permeating CPAs toward vitrification outcomes of mammalian tissues. In this review, we first discuss the roles of non-permeating CPAs including sugars and high molecular weight polymers in vitrification. We then summarize the supplemental effects of non-permeating CPAs on viability and functionalities of mammalian embryos, and ovarian, testicular, articular cartilage, tracheal, and kidney tissues following vitrification. Lastly, challenges associated with the use of non-permeating CPAs in vitrification of mammalian tissues are briefly discussed.


Effect of IGFBP6 Knockdown on Proteins Regulating Exosome Synthesis and Secretion in MDA-MB-231 Cell Line.
One of the potential causes of cancer recurrence is disruption of the cell-cell communication in the primary tumors that is realized, among other things, through secretion and uptake of exosomes by cells. Low expression of the IGFBP6 gene (insulin-like growth factor binding protein 6) is associated with a high recurrence rate and can serve as a prognostic marker of luminal breast cancer. The knockdown of the IGFBP6 gene leads to significant changes in lipid metabolism. We performed a quantitative analysis of both exosomes and proteins involved in the mechanism of their biogenesis. Changes in the expression profile of mRNAs and their proteins responsible for the synthesis and secretion of exosomes were revealed. We showed a decrease in the expression of the of the VPS28 gene mRNA (vacuolar protein sorting-associated protein 28) and the corresponding protein by 2.3 and 5.6 times, respectively. The secretion of exosomes by MDA-MB-231 cells with IGFBP6 knockdown decreased by 2 times. We discussed a mechanism of disruption of cell-cell communication.


Cerebrospinal fluid diversion devices and infection. A comprehensive review.
Hydrocephalus is a common neurosurgical disease affecting approximately 40 per 100,000 people. Cerebrospinal fluid diversion devices are essential in the management of this pathology. These devices include internal shunts and external ventricular drains. Infection is the most significant complication resulting from the high frequency of appearance and the consequences it involves, since it is associated with increased morbidity and mortality. It also involves increased hospital costs (approximately 40,000 euro per episode). In the present report the authors make a comprehensive review of cerebrospinal fluid diversion devices and their most important complication: infection. The authors make special emphasis on the epidemiology, aetiology, pathogenesis, risk factors, symptoms and signs, diagnosis, treatment and prevention of infection.


Effects of aging and sex on middle-latency somatosensory evoked potentials: normative data.
To investigate the exact relationship of middle-latency somatosensory evoked potential (SEP) components (in particular the third negative component N60) to age and sex in a normal population. Middle-latency SEPs are believed to show a considerable inter-individual variability and there is little published information on normative data of these components. Eighty-three healthy subjects aged 13-80 years were investigated using a standard SEP procedure. Both latency and amplitude of N60 increased substantially with age. Least square polynomial regression analysis showed that a quadratic curve best described its relationship (r2=0.67 for the latency, r2=0.18 for the amplitude). Data did not differ significantly with reference to sex or side of stimulation. The use of polynomial regression analysis for the determination of the normal range of N60 latencies and amplitudes has to be highly recommended. It might improve the accuracy with which abnormal results are detected in studies investigating the value of middle-latency SEPs in various diseases.


A pipeline of integrating transcriptome and interactome to elucidate central nodes in host-pathogens interactions.
Investigating the complexity of host-pathogen interactions is challenging. Here, we outline a pipeline to identify important proteins and signaling molecules in human-viral interactomes. Firstly, we curate a comprehensive human interactome. Subsequently, we infer viral targets and transcriptome-specific human interactomes (VTTSHI) for papillomavirus and herpes viruses by integrating viral targets and transcriptome data. Finally, we reveal the common and shared nodes and pathways in viral pathogenesis following network topology and pathway enrichment analyses. For complete details on the use and execution of this protocol, please refer to Kumar et al. (2020).


Examining the Association between Vitamin B12 Deficiency and Dementia in High-Risk Hospitalized Patients.
To explore the association between vitamin B12 deficiency and dementia in patients at high risk for vitamin B12 deficiency. Chart review. Emergency, critical care/ trauma, neurology, medicine, and rehabilitation units of two hospitals in Southwestern Ontario, Canada. Adult patients (n = 666) admitted from 2010 to 2012. Data collection included: reason for admission, gender, age, clinical signs and symptoms of B12 deficiency, serum B12 concentration, and B12 supplementation. Patients with dementia were identified based on their medication profile and medical history. Vitamin B12 deficiency (pmol/L) was defined as serum B12 concentration <148; marginal deficiency: ≥148-220 and adequate >220. Comparisons between B12-deficient patients with and without dementia were examined using parametric and non-parametric tests. Serum B12 values were available for 60% (399/666) of the patients, of whom 4% (16/399) were B12-deficient and 14% (57/399) were marginally deficient. Patients with dementia were not more likely to be B12-deficient or marginally deficient [21% (26/121)] compared to those with no dementia [17% (47/278), p=0.27)]. Based on documentation, 34% (25/73) of the B12-deficient and marginally-deficient patients did not receive B12 supplementation, of whom 40% (10/25) had dementia. In this sample of patients, there was no association between B12 deficiency and dementia. However, appropriate B12 screening protocols are necessary for high risk patient to identify deficiency and then receive B12 supplementation as needed.


Hypobetalipoproteinemic mice with a targeted apolipoprotein (Apo) B-27.6-specifying mutation: in vivo evidence for an important role of amino acids 1254-1744 of ApoB in lipid transport and metabolism of the apoB-containing lipoprotein.
Carboxyl-terminal deletion of apoB-100 may impair its triglyceride (TG)-transporting capability and alter its catabolism. Here, we compare our newly generated apoB gene (Apob)-targeted apoB-27.6-bearing mice to our previously reported apoB-38.9 mice to understand further the relationship between the size of a truncated apoB variant and its function/metabolism in vivo. The apoB-27.6-specifying mutation produces a premature stop codon six amino acids (aa) downstream of the last codon of mouse Apob exon 24 (corresponding to aa 1254 of human apoB-100). ApoB-27.6 transcripts were 3- and 5-fold more abundant than apoB wild type and apoB-38.9 transcripts in the liver. Likewise, hepatic secretion rates of apoB-27.6 were 7-fold higher than those of apoB-48 and apoB-38.9. In contrast, apoB-27.6 heterozygotes (Apob(27.6/+)) had lower hepatic TG secretion rates and higher liver TG contents than both apoB-38.9 heterozygotes (Apob(38.9/+)) and apoB wild type mice (Apob(+/+)). ApoB-27.6 was secreted by Apob(27.6/+) hepatocytes as dense high density lipoprotein particles. Moreover, despite its high secretion rates, apoB-27.6 was barely detectable in plasma. Disruption of apoE gene in Apob(38.9/+) and Apob(27.6/+) dramatically increased plasma levels of apoB-38.9 as well as apoB-48 but caused no change in plasma apoB-27.6 concentrations. Finally, the birth rate of apoB-27.6 homozygotes (Apob(27.6/27.6)) from intercrosses of Apob(27.6/+) was 7-fold lower than that of Apob(38.9/38.9) from Apob(38.9/+) intercrosses (1.8% versus 12%). Crossbreeding of Apob(27.6/27.6) and Apob(38.9/38.9) produced viable Apob(27.6/38.9) offspring, but Apob(27.6/27.6) intercrosses produced no offspring. Together, these results demonstrate in vivo that the apoB-27.6-apoB-38.9 peptide segment (aa 1254-1744) plays a critical role, not only in supporting hepatic TG-secretion and in modulating catabolism of apoB-containing lipoproteins, but also in normal mouse embryonic development.


Plasma concentration of rabeprazole after 8-week administration in gastroesophageal reflux disease patients and intragastric pH elevation.
The plasma concentration of rabeprazole in patients treated for gastroesophageal reflux disease (GERD) has not been reported, although the concentration in healthy volunteers has been reported previously. Here, CYP2C19 genotype effects of rabeprazole on the area under the plasma concentration-time curve (AUC) and the pH elevation were studied in GERD patients. Rabeprazole 10 mg/day was administrated for 8 weeks. AUC of rabeprazole in 18 Helicobacter pylori-negative GERD subjects (five CYP2C19 homozygous extensive metabolizers [homEM], eight heterozygous extensive metabolizers [hetEM] and five poor metabolizers [PM]) were determined after the final medication. Intragastric pH was recorded for 24 h at baseline and after the final medication. The AUC in the PM group (957 ng x h/mL) was significantly higher than those of homEM (375 ng x h/mL) and hetEM (542 ng x h/mL) groups. Median 24-h pH curves, median 24-h pH values and 24-h and nocturnal pH > 4 and pH > 3 holding times did not significantly differ among these groups. Disappearance of erosive lesions was observed after the treatment in all subjects with grade A, B or C at baseline irrespective of CYP2C19 genotypes. The AUC of rabeprazole depended on the CYP2C19 genotypes in Japanese GERD patients; however, the intragastric pH elevation was independent of CYP2C19 genotypes, which is consistent with the CYP2C19 genotype-independent healing efficacy of erosive lesions by rabeprazole. The present low AUC values indicated that abnormal accumulative effects on AUC did not occur during the period of the 8-week administration of rabeprazole.


Subchondral plate porosity colocalizes with the point of mechanical load during ambulation in a rat knee model of post-traumatic osteoarthritis.
This study investigated the association between spatiotemporal cartilage-subchondral bone plate alterations and mechanical load during ambulation in an experimental rat model of destabilized medial meniscus (DMM). Twelve-week-old Wistar rats (n = 38) underwent DMM surgery on the right knee and sham surgery on the left knee. At 2 and 4 weeks after surgery, subchondral bone changes were evaluated via micro-computed tomography with various knee flexion angles to simulate weight-bearing during rat ambulation under a 3-dimensional motion capture apparatus. Additionally, the biomechanical properties, histology, and ultrastructure of the medial tibia and femoral condyle were evaluated. Focal subchondral bone plate perforations were confirmed in the medial tibia within 2 weeks after surgery and were aggravated rapidly 2 weeks later. This subchondral plate porosity colocalized with articular cartilage lesions as confirmed by histology and scanning electron microscopy, and coincided with the likely point of contact between the posterior femoral condyle and tibial plateau during ambulation. Biomechanical properties were confirmed at the medial tibia, at which stiffness was reduced to approximately half that of the sham-operated knee at 4 weeks after surgery. Cartilage-subchondral bone plate alterations localized in the region of the point of mechanical load during ambulation in DMM-operated knees, at which the mechanical integrity of cartilage was impaired. These results indicate that DMM-induced increases in mechanical load play an important role in the pathogenesis of early post-traumatic osteoarthritis (OA), and it might accelerate the development of the disease via cartilage-subchondral bone plate crosstalk through increased subchondral plate perforations.


Prediction of functional results of percutaneous coronary interventions with virtual stenting and quantitative flow ratio.
The clinical value of residual quantitative flow ratio (rQFR), a novel function of QFR technique, is unknown. We investigated the clinical value of rQFR, aimed to predict residual ischemia after virtual percutaneous coronary intervention (vPCI). This is a substudy of the COE-PERSPECTIVE registry, which investigated the prognostic value of post-PCI fractional flow reserve (FFR). From pre-PCI angiograms, QFR and rQFR were analyzed and their diagnostic performance was assessed at blinded fashion using pre-PCI FFR and post-PCI FFR as reference, respectively. The prognostic value of rQFR after vPCI was assessed according to vessel-oriented composite outcome (VOCO) at 2 years. We analyzed 274 patients (274 vessels) with FFR-based ischemic causing lesions (49%) from 555 screened patients. Pre-PCI QFR and FFR were 0.63 ± 0.10 and 0.66 ± 0.11 (R = 0.756, p < 0.001). rQFR after vPCI and FFR after real PCI were 0.93 ± 0.06 and 0.86 ± 0.07 (R = 0.528, p < 0.001). The mean difference between rQFR and post-PCI FFR was 0.068 (95% limit of agreement: -0.05 to 0.19). Diagnostic performance of rQFR to predict residual ischemia after PCI was good (area under the curve [AUC]: 0.856 [0.804-0.909], p < 0.001). rQFR predicted well the incidence of 2-year VOCO after index PCI (AUC: 0.712 [0.555-0.869], p = 0.041), being similar to that of actual post-PCI FFR (AUC: 0.691 [0.512-0.870], p = 0.061). rQFR ≤0.89 was associated with increased risk of 2-year VOCO (hazard ratio [HR]: 12.9 [2.32-71.3], p = 0.0035). This difference was mainly driven by a higher rate of target vessel revascularization (HR: 16.98 [2.33-123.29], p = 0.0051). rQFR estimated from pre-PCI angiography and virtual coronary stenting mildly overestimated functional benefit of PCI. However, it well predicted suboptimal functional result and long-term vessel-related clinical events. Influence of fractional flow reserve on the Clinical OutcomEs of PERcutaneouS Coronary Intervention (COE-PESPECTIVE) Registry, NCT01873560.


Effect of disease modifying antirheumatic drugs and nonsteroidal antiinflammatory drugs upon cellular and fibronectin responses in a pleurisy model.
Intrapleural injections of carrageenan into rats resulted in peak increases in intrapleural inflammatory cell counts (90% mononuclear cells) and fibronectin content at 3 days after the injections. Administration of disease modifying antirheumatic drugs (DMARD) prevented the increases in both fibronectin levels and intrapleural cell counts, whereas nonsteroidal antiinflammatory drugs, in general, potentiated the increases. Inhibition of fibronectin production by DMARD in carrageenan induced pleurisy in rats may be related to the antirheumatic activity of this class of drugs.


Antibacterial activity of Thymus maroccanus and Thymus broussonetii essential oils against nosocomial infection - bacteria and their synergistic potential with antibiotics.
The aim of this study was to evaluate the antibacterial effect of the association between conventional antibiotics and essential oils (EOs) of endemic Moroccan thyme species, Thymus maroccanus and T. broussonetii, on antibiotic-resistant bacteria involved in nosocomial infections. Synergistic interactions between antibiotics (ciprofloxacin, gentamicin, pristinamycin, and cefixime) and EOs, and between T. maroccanus and T. Broussonetii EOs were determined by the checkerboard test. Serial dilutions of two antimicrobial agents were mixed together so that each row (and column) contained a fixed amount of the first agent and increasing amounts of the second one. The results indicate that the oils had a high inhibitory activity against tested bacteria, except for Pseudomonas aeruginosa. In parallel with the increase of cellular killing, the release of 260nm-absorbing materials from bacterial cells, treated with EOs, increased in response to oil concentration. Out of 80 combinations tested between EOs and antibiotics, 71% showed total synergism, 20% had partial synergistic interaction and 9% showed no effect. Combination with carvacrol, the major constituent of T. maroccanus and T. broussonetii, showed also an interesting synergistic effect in combination with ciprofloxacin. The effect on Gram-positive bacteria was more important than on Gram-negative bacteria. These findings are very promising since the use of these combinations for nosocomial infections treatment is likely to reduce the minimum effective dose of the antibiotics, thus minimizing their possible toxic side effects and treatment cost. However, further investigations are needed to assess the potential for therapeutic application.


Effective risk management in the shadow of COVID-19 pandemic: The evidence of Indonesian listed corporations.
The study uses COVID-19 to identify the treatment group as the difference in change of non-financial corporations (NFCs) risk management ratios over time to investigate the causal effect of the NFCs' effective risk management (ERM) practices on operational efficiency (OE). ERM was measured by solvency and liquidity ratios, while the risk management theory was developed to refine the scope of the study. The data were collected from the central bank of Indonesia to map the empirical analysis, and the difference in difference (DID) technique was used to illustrate how NFCs react to mitigate the negative impact of COVID-19 and generate OE. Specifically, a quasi-natural experiment was used to size the effect of ERM practices on corporate OE during the COVID-19 pandemic. The descriptive analysis revealed that the COVID-19 pandemic effect has been unequal across different industrial sectors. Moreover, the empirical findings showed that corporate risk management during COVID-19 is the source of structural change, which affects its existence and operational efficiency. While debt amount and age may affect corporate credit score, ERM practices led the indebted corporation to the flexibility of debt refinancing or/and restructuring, which offers them the ability to prevent bankruptcy and adapt to the changes while operating efficiently. The finding revealed evidence of the important role of long-term debt in offering protection to NFCs during the credit supply shock brought in by the COVID-19 pandemic. Furthermore, the findings show that long-term debt is negatively associated with corporate OE. This was expected given that corporations use long-term debt financing for long-term investment, while short-term debt funds the working capital. Thus, to assess the effect of debts on corporate OE, managers should consider their maturity structure, among other factors.


The Cartesian clock metaphor for pineal gland operation pervades the origin of modern chronobiology.
In theoretical descriptions formulated during the 1600s, R. Descartes attributed a clock-like role to the pineal gland. He established the belief that pineal function underlies the laws of the universe that determine the cyclic sleep-awake states in man. Recent reports about pineal circadian pacemakers now validate the brilliant accuracy of Cartesian thought, in relation to the relevant role of the pineal gland.


The FOXO1 Transcription Factor Instructs the Germinal Center Dark Zone Program.
The pathways regulating formation of the germinal center (GC) dark zone (DZ) and light zone (LZ) are unknown. In this study we show that FOXO1 transcription factor expression was restricted to the GC DZ and was required for DZ formation, since its absence in mice led to the loss of DZ gene programs and the formation of LZ-only GCs. FOXO1-negative GC B cells displayed normal somatic hypermutation but defective affinity maturation and class switch recombination. The function of FOXO1 in sustaining the DZ program involved the trans-activation of the chemokine receptor CXCR4, and cooperation with the BCL6 transcription factor in the trans-repression of genes involved in immune activation, DNA repair, and plasma cell differentiation. These results also have implications for the role of FOXO1 in lymphomagenesis because they suggest that constitutive FOXO1 activity might be required for the oncogenic activity of deregulated BCL6 expression.


Identifying and validating key genes mediating intracranial aneurysm rupture using weighted correlation network analysis and exploration of personalized treatment.
Intracranial aneurysmal subarachnoid hemorrhage (aSAH) is a dangerous and highly fatal condition if ruptured. Significant advances have been made in the treatment of unruptured intracranial aneurysms (UIAs), but risk assessment methods for early diagnosis of intracranial aneurysm (IA) rupture remain limited. The datasets of IA GSE13353, GSE15629, and GSE54083 were downloaded through the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) in unruptured and ruptured aneurysms were identified by R software using methods such as gene set enrichment analysis (GSEA) and weighted gene co-expression network analysis (WGCNA). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were performed on the DEGs, and logistic regression models were used to construct a prediction model to discriminate UIA from healthy samples. We then performed GSEA on the genes in the model, followed by model validation using the GSE54083 dataset. Finally, we used the single-sample (ss)GSEA method to investigate the relationship between the diagnostic model genes and immune cells and immune function. A total of 79 DEGs were obtained in patients with IA rupture compared to unruptured controls. The results of KEGG and GO enrichment analysis showed that neutrophil activation is involved in immune response, neutrophil mediated immunity, and positive regulation of angiogenesis. Interestingly, the results of immunoassays demonstrated that the break in IA may be associated with immune T cells. We used DEGs and WGCNA to determine common genes. The logistic regression model was trained based on 24 intersecting genes, and eventually retained 2 genes, KIAA0226L and UPP1, which we found to be reliable using the validation set, and GSEA revealed that the diagnostic model was associated with the Hippo signaling pathway and vascular smooth muscle contraction, and viral protein interaction with cytokine and cytokine were also associated. Finally explored using the CMap database, Tivozanib could be a potential small molecule drug for the treatment of ruptured intracranial aneurysms (RIAs). We identified new diagnostic genes associated with IA rupture, which may provide a new way of aneurysm diagnosis.


Declining COVID-19 case-fatality in Georgia, USA, March 2020 to March 2021: a sign of real improvement or a broadening epidemic?
To examine whether declines in the crude U.S. COVID-19 case fatality ratio is due to improved clinical care and/or other factors. We used multivariable logistic regression, adjusted for age and other individual-level characteristics, to examine associations between report month and mortality among confirmed and probable COVID-19 cases and hospitalized cases in Georgia reported March 2, 2020 to March 31, 2021. Compared to August 2020, mortality risk among cases was lowest in November 2020 (OR = 0.84; 95% CI: 0.78-0.91) and remained lower until March 2021 (OR = 0.86; 95% CI: 0.77-0.95). Among hospitalized cases, mortality risk increased in December 2020 (OR = 1.16, 95% CI: 1.07-1.27) and January 2021 (OR = 1.25; 95% CI: 1.14-1.36), before declining until March 2021 (OR = 0.90, 95% CI: 0.78-1.04). After adjusting for other factors, including the shift to a younger age distribution of cases, we observed lower mortality risk from November 2020 to March 2021 compared to August 2020 among cases. This suggests that improved clinical management may have contributed to lower mortality risk. Among hospitalized cases, mortality risk increased again in December 2020 and January 2021, but then decreased to a risk similar to that among all cases by March 2021.


Idiopathic pulmonary fibrosis--a systematic review on methodology for the collection of epidemiological data.
Recent studies suggest that the incidence of idiopathic pulmonary fibrosis (IPF) is rising. Accurate epidemiological data on IPF, however, are sparse and the results of previous studies are contradictory. This study was undertaken to gain insight into the various methods used in the epidemiological research of IPF, and to get accurate and comparable data on these different methodologies. A systematic database search was performed in order to identify all epidemiological studies on IPF after the previous guidelines for diagnosis and treatment were published in 2000. Medline (via Pubmed), Science Sitation Index (via Web of Science) and Embase databases were searched for original epidemiological articles published in English in international peer-reviewed journals starting from 2001. After pre-screening and a full-text review, 13 articles were accepted for data abstraction. Three different methodologies of epidemiological studies were most commonly used, namely: (1) national registry databases, (2) questionnaire-based studies, and (3) analysis of the health care system's own registry databases. The overall prevalence and incidence of IPF varied in these studies between 0.5-27.9/100,000 and 0.22-8.8/100,000, respectively. According to four studies the mortality and incidence of IPF are rising. We conclude that there are numerous ways to execute epidemiological research in the field of IPF. This review offers the possibility to compare the different methodologies that have been used, and this information could form a basis for future studies investigating the prevalence and incidence of IPF.


[Protein S, C and antithrombin deficiency: association with myocardial infarction and thromboembolism in the young].
The pathogenesis of myocardial infarction (MI) in young involves new factors including constitutional or acquired thrombophilia. To determine in patients ² 50 years, the association between coagulation factors deficiency, myocardial infarction and cardiovascular events during follow-up. Protein C (PC), PS and antithrombin (AT) were screened in 50 patients admitted for acute MI and in a healthy control group. Univariate and multivariate analysis were performed using SPSS 11.5 version. PS and PC deficiency were associated to MI (respectively 24% vs 0%, p=0.001 and 14% vs 0%, p=0.016), independently for PC. No AT deficiency was detected in both groups. During followup, PS and C deficiency were predictive for venous thrombosis (p<0.05) and PS deficiency for pulmonary embolism. Protein C and S deficiency may play an important role in MI in young and also in thromboembolic complications during follow-up. Nevertheless, therapeutic implications remain controversial.


Evolution of the amino acid code: inferences from mitochondrial codes.
The amino acid code is usually presented as a table of 64 codons. Actually the code results from the action of tRNA molecules that carry amino acids to codons in mRNA by means of codon-anticodon pairing. The tRNA molecules are transcribed from genes that undergo evolution and the number of anticodons can therefore increase during evolution, but the number of codons is fixed at 64. Mammalian mitochondrial codes contain only 22 anticodons for 20 amino acids as compared with 54 anticodons for 20 amino acids in the universal code. It is proposed that an archetypal code containing 16 anticodons for 15 amino acids evolved into the universal code by gene duplication, followed by mutations that modified the anticodons and amino acid acceptor sites. In substantiation of this proposal, it is noted that the mammalian mitochondrial code is simplified by comparison with the universal code. For example, single anticodons are used for each of eight amino acids in the mammalian mitochondrial code. This simplification may represent an evolutionary retrogression towards the proposed archetypal code.


Stepwise molecular mechanisms responsible for chemoresistance in bladder cancer cells.
Chemotherapy resistance is an obstacle to cancer therapy and is considered a major cause of recurrence. Thus, understanding the mechanisms of chemoresistance is critical to improving the prognosis of patients. Here, we have established a stepwise gemcitabine-resistant T24 bladder cancer cell line to understand the molecular mechanisms of chemoresistance within cancer cells. The characteristics of the stepwise chemoresistance cell line were divided into 4 phases (parental, early, intermediate, and late phases). These four phase cells showed increasingly aggressive phenotypes in vitro and in vivo experiments with increasing phases and revealed the molecular properties of the biological process from parent cells to phased gemcitabine-resistant cell line (GRC). Taken together, through the analysis of gene expression profile data, we have characterized gene set of each phase indicating the response to anticancer drug treatment. Specifically, we identified a multigene signature (23 genes including GATA3, APOBEC3G, NT5E, MYC, STC1, FOXD1, SMAD9) and developed a chemoresistance score consisting of that could predict eventual responsiveness to gemcitabine treatment. Our data will contribute to predicting chemoresistance and improving the prognosis of bladder cancer patients.


Reducing vaccine hesitancy in pregnancy by the health literacy model inclusive of modern communication tools.
Albeit the vaccination is one of the most successful and cost-effective public health interventions, reluctance or refusal to vaccinate represents one of the ten threats to global health by the World Health Organization. Additional efforts to promote vaccination are required for at higher risk categories, such as pregnant women. Our approach supports the role of a clear and transparent communication by using efficient interventions and educational strategies to increase both willingness and confidence for preventable diseases in neonates and infants by vaccinations in pregnancy.


Laparoscopic ventral rectopexy for obstructed defecation syndrome.
Obstructed defecation remains a serious syndrome. Several procedures have been applied to treat it. A concomitant enterocele excludes some of these procedures, because of potential threat of damaging the bowel. The aim of this study was to assess the outcome of patients who underwent laparoscopic nerve sparing ventral rectopexy for obstructed defecation syndrome with concomitant enterocele. Seventeen patients were included. Data about clinical history, physical examination and a defecogram were collected. All patients underwent a laparoscopic ventral rectopexy. Complications, hospital stay, postoperative morbidity and long-term outcome were documented. All patients underwent laparoscopic ventral rectopexy. The median operating time was 199 min (range 186-239 min). One conversion laparotomy was required. Six patients had postoperative complications (ileus n = 2, posttraumatic leg dystrophy n = 1, wound infection n = 1, incisional hernia n = 2). The median hospital stay was 6 days (range 3-24 days). Fifteen patients had improvement of their defecation problem, although six patients still had minor constipation symptoms. In one patient the mesh was rejected and finally removed. Obstructed defecation syndrome is a combined functional and mechanical problem. In selected patients, especially when an enterocele is present, laparoscopic ventral rectopexy is a feasible technique, with an acceptable number of complications.


Bombesin microinfusion into the paraventricular nucleus suppresses gastric acid secretion in the rat.
Bombesin is a particularly potent inhibitor of gastric acid secretion when injected intracisternally in the rat. Because bombesin-like immunoreactivity is found in several forebrain regions implicated in gut regulation, the ability of bombesin to affect gastric secretion was tested in these areas by direct microinfusion. Bombesin significantly and dose-relatedly suppressed gastric acid secretion when it was infused into the hypothalamic paraventricular nucleus. Bombesin microinfusion into the ventromedial or lateral hypothalamic areas, or the caudate-putamen, had no significant effect. A further experiment using glass micropipets showed that back-diffusion of bombesin along the cannula track to a distant site of action was unlikely to account for the results obtained, and provided further evidence that the active site is limited to the paraventricular nucleus and possibly the ventralmost nucleus reuniens. The results suggest that the bombesin receptors and immunoreactive terminals previously identified in this region may be involved in the central regulation of gastric secretion.


Defining the oncogenic function of the TEL/AML1 (ETV6/RUNX1) fusion protein in a mouse model.
The t(12;21) translocation, generating the TEL/AML1 fusion protein, is the most common genetic lesion in childhood cancer. Using a bone marrow transplantation model, we demonstrate that TEL/AML1 expression impinges on normal hematopoietic differentiation, leading to the in vivo accumulation and persistence of an early progenitor compartment with a Sca1(+)/Kit(hi)/CD11b(+) phenotype and an increased self-renewal capacity, as documented by replating assays in vitro. Differentiation of these cells is not blocked, but the frequency of mature blood cells arising from TEL/AML1-transduced progenitors is low. Impaired differentiation is prominently observed in the pro-B-cell compartment, resulting in an proportional increase in early progenitors in vivo, consistent with the t(12;21) ALL phenotype. Despite the accumulation of both multipotent and B-cell progenitors in vivo, no leukemia induction was observed during an observation period of over 1 year. These results are consistent with findings in twins with concordant ALL, showing that TEL/AML1 generates a preleukemic clone in utero that persists for several years in a clinically covert fashion. Furthermore, our studies showed that the pointed domain of TEL/AML1, which recruits transcriptional repressors and directs oligomerization with either TEL/AML1 or wild-type TEL, was essential for the observed differentiation impairment and could not be replaced with another oligomerization domain.


Perceptions of nursing as a career choice of students in the Baccalaureate nursing program.
Schools of nursing must recruit and retain qualified applicants in order to confront the current challenge to nursing resources. Perceptions of nursing have been linked to students' decisions to enter the nursing profession and to continue in or withdraw from nursing programs. As part of an on-going longitudinal study in Canada, the nursing attitude questionnaire and nursing orientation tool were used to explore the perceptions of nursing of 213 beginning and 150 graduating students in a Baccalaureate nursing program. Overall, both groups of students held a positive image of nursing. There were however, significant differences between the groups in their orientation to nursing as well as their views on nursing roles, education, political issues, and the value of nursing as a profession. Implications for recruitment of nursing students are presented.


Guide to Health: A Randomized Controlled Trial of the Effects of a Completely WEB-Based Intervention on Physical Activity, Fruit and Vegetable Consumption, and Body Weight.
Theory-based, efficacious, long-term, completely Internet-based interventions are needed to induce favorable shifts in health behaviors and prevent weight gain. To assess nutrition, physical activity, and, secondarily, body weight outcomes in the tailored, social cognitive theory Guide to Health (WB-GTH) program with all recruitment, assessment, and intervention performed on the Internet. The focus of the efficacy study was engaged participants who completed 3 or more program modules plus baseline, 6-months post and, 16-months follow-up assessments (n = 247). To be eligible, participants needed to be between 18-63 years of age, with a BMI between 23-39, sedentary to low-active but otherwise healthy. Participant had a mean age of 45.5 years (10.3), 86.2% were female, with 8.5% from minority groups, with a mean 17.5 (3.0) years of education, and had a median annual household income of about $85k. Nevertheless, about 83% were overweight or obese and about 75% were sedentary (i.e., <5000 steps/day) or had low levels of activity (i.e., 5,000 - 7499 steps/day). Participants were randomized to the WB-GTH-Basic intervention or WB-GTH-Enhanced intervention. Content, overall target behaviors, program goals and strategies were the same in the two interventions with the difference that Basic included a generic feedback and planning approach and Enhanced included a highly tailored planning and feedback approach. Participants reported at assessments pedometer step counts to assess physical activity, bodyweight from a scale provided, and fruit and vegetable (F&V) servings were assessed from food frequency questionnaires completed online. Participants in both Basic and Enhanced at follow-up increased physical activity by about 1400 steps/day, lost about 3% of bodyweight, and increased F&V by about 1.5 serving/day. There was evidence that the least physically active, those who were obese, and those with poorest nutrition made greater long-term improvements. Given similar outcomes for Basic and Enhanced, a relatively simple entirely Internet-based program can help people improve health behaviors and prevent weight gain.


Hepatitis B Virus Infection: Overview.
Hepatitis B virus (HBV) is a DNA virus, belonging to the Hepadnaviridae family. It is a partially double-stranded DNA virus with a small viral genome (3.2 kb). Chronic HBV infection remains a global public health problem. If left untreated, chronic HBV infection can progress to end-stage liver disease, such as liver cirrhosis and hepatocellular carcinoma (HCC). In recent years, tremendous advances in the field of HBV basic and clinical research have been achieved, ranging from the HBV biological characteristics, immunopathogenesis, and animal models to the development of new therapeutic strategies and new drugs against HBV. In this overview, we begin with a brief history of HBV discovery and treatment milestones. We then briefly summarize the HBV research advances, which will be detailed in the following chapters.


Swertiamarin or heat-transformed products alleviated APAP-induced hepatotoxicity via modulation of apoptotic and Nrf-2/NF-κB pathways.
Swertiamarin (STM) belongs to iridoid class of compounds, and the heat-transformed products (HTPS) are produced by STM in the process of drug processing. The purpose of this study was to explore the protective effect and mechanism of STM or HTPS on acetaminophen (APAP)-induced hepatotoxicity. Mice and L-O2 cells were given APAP to establish the hepatotoxicity model in vivo and in vitro. The effects of STM or HTPS on oxidative stress, inflammation, and apoptosis induced by APAP were evaluated, with N-acetylcysteine (NAC) as a positive control. STM or HTPS reduced the APAP-induced apoptosis of L-O2 cells and significantly alleviated the liver injury index induced by APAP (p < 0.01, 0.005) Interestingly, HTPS had better protective effect against APAP-induced hepatotoxicity than STM (p < 0.05). In addition STM or HTPS improved the histological abnormalities; inhibited lipid peroxidation and reduced the level of inflammatory mediators. They also activated the defense system of nuclear factor erythroid 2 related factor 2 (Nrf-2) and inhibited nuclear factor-κ B (NF-κB).