Stimulatory effect of thapsigargin, a non-TPA-type tumor promoter, on arachidonic acid metabolism in the murine keratinocyte line HEL30 and on epidermal cell proliferation in vivo as compared to the effects of phorbol ester TPA.
The effect of thapsigargin (Tg), a non-12-O-tetradecanoylphorbol-13-acetate (TPA) type skin tumor promoter, on arachidonic acid and prostaglandin E2 (PGE2) formation in HEL30 keratinocytes and on epidermal DNA synthesis in vitro and in vivo (mouse skin) was investigated and compared with that of the phorbol ester TPA. On a molar basis Tg was 30-fold more potent in inducing the arachidonic acid/PGE2 release than TPA. Applied together, the two agents showed a strong synergistic action. The response critically depended on the presence of Ca2+ in the extracellular medium. While the TPA-induced release was mediated by protein kinase C (PKC) the Tg-induced release was not. In contrast to TPA (1 microM), which is a stimulator of HEL30 DNA synthesis, Tg (0.1-1 microM) inhibited DNA labeling in vitro due to a pronounced cytotoxic effect. TPA did not exhibit such an effect. In vivo both agents were practically equipotent in inducing epidermal DNA synthesis and hyperplasia with TPA having an approximately 10-fold higher irritating potential than Tg. It is concluded that the hyperplasiogenic and tumor-promoting effect of Tg in vivo is due to cytotoxicity causing cell death and regenerative hyperproliferation. Thus, Tg-induced skin tumor promotion seems to resemble tumor promotion by mechanical skin wounding, whereas TPA evokes a more specific, i.e. PKC-mediated response. Since despite these mechanistic differences both agents induce an immediate release of arachidonic acid/PGE2 in keratinocytes, this response may be considered to provide an in vitro parameter for irritancy and tumor promotion.


Biosynthesis of biotin-vitamers from unsaturated higher fatty acids by bacteria.
The biosynthesis of biotin-vitamers from unsaturated higher fatty acids by the resting cell system of bacteria was investigated. Biotin-vitamer formation from oleic, linoleic and linolenic acids was found in certain bacteria belonging to the genera Enterobacter, Agrobacterium, Bacillus, Escherichia and Micrococcus. The biotin-vitamers were identified as a mixture of 7,8-diaminopelargonic acid, desthiobiotin and biotin. Metabolites other than biotin-vitamers from those fatty acids were analyzed by gas-liquid chromatography and mass spectrometry. Linoleic, oleic, C-17 and C-15 saturated fatty acids were identified as the main metabolites of linolenic acid. The pathway of biosynthesis of pimelic acid from linolenic acid by a strain of Bacillus sphaericus AKU 0227 was also studied.


Design, synthesis, and biological evaluation of structurally modified isoindolinone and quinazolinone derivatives as hedgehog pathway inhibitors.
The Hedgehog (Hh) signaling pathway is associated with diverse aspects of cellular events, such as cell migration, proliferation, and differentiation throughout embryonic development and tissue patterning. An abnormal Hh signaling pathway is linked to numerous human cancers, including basal cell carcinoma (BCC), medulloblastoma (MB), lung cancer, prostate cancer, and ovarian cancer, and it is therefore a promising target in cancer therapy. Using a structure-hopping approach, we designed new Hh signaling pathway inhibitors with isoindolinone or quinazolinone moieties, which were synthesized and biologically evaluated using an 8xGli-luciferase (Gli-Luc) reporter assay in NIH3T3 cells. Compounds 9-11 and 14 with isoindolinone scaffolds demonstrated moderate Hh inhibitory activity; whereas quinazolinone derivatives 24, 29, 32, 34, and 35 exhibited good potency with submicromolar IC50 values and the analog 28 showed nanomolar IC50 value. Although sonidegib shows a decrease in inhibitory effect on vismodegib resistance-conferring Smo mutants, the structurally modified new compounds not only possess the pharmacophoric properties of Hh pathway inhibition but also preserve the suppressive potency in drug-resistant Smo mutants. Mechanistically, quinazolinone derivatives 28 and 34 suppress Hh signaling by blocking Smo and Gli translocation into the cilia, similar to vismodegib and sonidegib. Additionally, the human microsomal stability of the representative analogs 28 and 34 were determined to be comparable to that of the reference compound sonidegib. Thus, these new scaffolds can serve as a platform for the development of novel cancer therapeutics targeting the Hh pathway.


ISGylation is increased in the peripheral blood mononuclear cells derived from symptomatic COVID-19 patients.
Cytokine-driven hyper inflammation has been identified as a critical factor behind poor outcomes in patients severely infected with SARS-CoV-2 virus. Notably, protein ISGylation, a protein conjugated form of Type 1 IFN-inducible ubiquitin-like protein ISG15 (Interferon-Stimulated Gene 15), induces cytokine storm (CS) and augments colonic inflammation in colitis-associated colon cancers in mouse models. However, whether ISGylation is increased and causally responsible for CS and hyper inflammation in symptomatic COVID-19 patients is unknown. Here, we measured ISGylation levels in peripheral blood mononuclear cells (PBMCs) from 10 symptomatic (SARS-CoV-2-positive with symptoms) and asymptomatic (SARS-CoV-2-positive with no symptoms) COVID-19 patients, and 4 uninfected individuals (SARS-CoV-2-negative), using WesTm assay. Strikingly, we note significant increases in protein ISGylation and MX-1 (myxovirus-resistance protein-1) protein levels, both induced by type-I IFN, in symptomatic but not in asymptomatic patients and uninfected individuals. Knowing that ISGylation augments CS and intestinal inflammation in colon cancers, we propose that increased ISGylation may be an underlying cause of CS and inflammation in symptomatic patients.


Joint Torque and Mechanical Power of Lower Extremity and Its Relevance to Hamstring Strain during Sprint Running.
The aim of this study was to quantify the contributions of lower extremity joint torques and the mechanical power of lower extremity muscle groups to further elucidate the loadings on hamstring and the mechanics of its injury. Eight national-level male sprinters performed maximum-velocity sprint running on a synthetic track. The 3D kinematic data and ground reaction force (GRF) were collected synchronously. Intersegmental dynamics approach was used to analyze the lower extremity joint torques and power changes in the lower extremity joint muscle groups. During sprinting, the GRF during the stance phase and the motion-dependent torques (MDT) during the swing phase had a major effect on the lower extremity movements and muscle groups. Specifically, during the stance phase, torque produced and work performed by the hip and knee muscles were generally used to counteract the GRF. During the swing phase, the role of the muscle torque changed to mainly counteract the effect of MDT to control the movement direction of the lower extremity. Meanwhile, during the initial stance and late swing phases, the passive torques, namely, the ground reaction torques and MDT produced by the GRF and the inertial movement of the segments of the lower extremity, applied greater stress to the hamstring muscles.


HIPK2 Cooperates with KRAS Signaling and Associates with Colorectal Cancer Progression.
Homeodomain-interacting protein kinase 2 (HIPK2) is an evolutionary conserved kinase that has gained attention as a fine tuner of multiple signaling pathways, among which those commonly altered in colorectal cancer. The aim of this study was to evaluate the relationship of HIPK2 expression with progression markers and mutational pattern and gain insights into the contribution of HIPK2 activity in colorectal cancer. We evaluated a retrospective cohort of colorectal cancer samples by IHC for HIPK2 expression and by next-generation sequencing (NGS) for the detection of mutations of cancer associated genes. We show that the percentage of HIPK2-positive cells increases with tumor progression, significantly correlates with tumor-node-metastasis (TNM) staging and associates with a worse outcome. In addition, we observed that high HIPK2 expression significantly associates with KRAS mutations but not with other cancer-related genes. Functional characterization of the link between HIPK2 and KRAS show that activation of the RAS pathway either due to KRAS mutation or via upstream receptor stimulation, increases HIPK2 expression at the protein level. Of note, HIPK2 physically participates in the active RAS complex while HIPK2 depletion impairs ERK phosphorylation and the growth of tumors derived from KRAS mutated colorectal cancer cells. Overall, this study identifies HIPK2 as a prognostic biomarker candidate in patients with colorectal cancer and underscores a previously unknown functional link between HIPK2 and the KRAS signaling pathway. Our data indicate HIPK2 as a new player in the complex picture of the KRAS signaling network, providing rationales for future clinical studies and new treatment strategies for KRAS mutated colorectal cancer.


Prevention of vasospasm by clot removal and intrathecal bolus injection of tissue-type plasminogen activator: preliminary report.
In this study, we evaluated the efficacy of postoperative intrathecal injections of tissue-type plasminogen activator (tPA) in preventing cerebral vasospasm in cases with a diffuse severe subarachnoid hemorrhage. All 10 cases were graded Group 3 according to the classification of Fisher and associates, and the CT number (Hounsfield number) of the subarachnoid clot was over 75. After clipping the aneurysm and removing the clot, three cisternal drainage catheters were inserted into both sylvian cisterns and the prepontine cistern, and continuous ventricular drainage was performed routinely. Postoperatively, tPA (0.5 mg/2.5 ml) was infused as a bolus into both basal cisterns and the lateral ventricle twice daily for about 6 days. Angiography and cerebral blood flow studies using single photon emission computed tomography were performed on Day 4 or 5 and between Days 7 and 10 after onset of the hemorrhage. To date, there have been no cases that have shown angiographic vasospasm or delayed ischemic neurological deficits. This preliminary study indicates that the intrathecal bolus injection of tPA produces a marked effect on vasospasm.


Randomized trial of 5-fluorouracil and high-dose folinic acid with or without alpha-2B interferon in advanced colorectal cancer.
We evaluated the role of low-dose alpha-2b interferon, added to chemotherapy, for advanced colorectal cancer; we randomized patients, to either a combination chemotherapy of 5-fluorouracil (5-FU) and high-dose folinic acid (HDFA) or the same regimen plus interferon. Between January 1990 and March 1992, 100 untreated patients (PTS) with advanced colorectal cancer, 53 men and 47 women, with an ECOG performance status (PS) of < or = 3, were randomized to either HDFA 200 mg/m2 iv bolus and 5FU 370 mg/m2 in 15-min iv infusion days 1-5 every 4 weeks (arm A), or the same chemotherapy plus IFN 3 x 10(6) IU subcutaneously three times a week in chemotherapy intervals (arm B). A total of 97 PTS are evaluable for response, toxicity, and survival; 3 PTS are not evaluable in arm B for major protocol violations. PTS characteristics were well balanced in both arms for age (median, 64 years), disease-free survival, and disease site. ECOG PS was 0 in 28% of PTS in arm A and in 13% in arm B. Response rates were as follows: arm A, 40%; and arm B, 23%. Median time to failure was as follows: 10.2 months arm A versus 9 months arm B. Median survival was as follows: 13.3 months arm A versus 10.9 months arm B. Grade 3 haematological toxicity was 9% of PTS in both arms. Gastrointestinal toxicity was as follows: 17% arm A versus 22% arm B. The cost of drugs expressed per m2/month was $60 in arm A and $390 in arm B. The results show that IFN at the schedule and doses employed adds no benefit to the combination of 5FU/HDFA.


Highly selective vagotomy with dilatation or duodenoplasty. A surgical alternative for obstructing duodenal ulcer.
Highly selective vagotomy (HSV) is an accepted choice for the treatment of uncomplicated duodenal ulcer. Its use in patients with gastric outlet obstruction, however, remains quite controversial. Since 1980, 69 patients have undergone HSV at the Medical College of Georgia Hospitals. Of these, 20 (29%) underwent either dilatation (14) or duodenoplasty (6) for accompanying outlet obstruction. The obstruction was graded as severe in 17 (85%) and moderate in three (15%). Follow-up evaluation has included Visick grading and endoscopy. There have been two deaths (38 and 54 months following surgery). Both patients were Visick I. Of the 18 patients available for review to date, 12 (67%) are Visick I and four (22%) are Visick II. There have been two failures (11%), discovered only by endoscopic follow-up in asymptomatic patients. No patients have required reoperation. HSV with dilatation or duodenoplasty is a reasonable surgical alternative for the treatment of obstructing duodenal ulcer disease.


Disparities in identification of comorbid diagnoses in children with ADHD.
This study explores disparities in identification of educationally relevant comorbidities and medication prescribing practices for children with attention-deficit hyperactivity disorder (ADHD) and either comprehensive neurodevelopmental evaluations or evaluations limited by insurance to behavior management with medication. This study was a retrospective chart review of 5- to 10-year-old children with ADHD diagnosed at the initial evaluation. Data collected included demographics, rates of comorbid conditions, medication management, and educational interventions. The 2 groups were similar in age, educational supports, and medication management. The group with insurance permitting comprehensive evaluations was more likely to be Caucasian, have higher parental education levels, and have more comorbid conditions identified with academic impact. School-aged children with ADHD are likely to receive similar educational and medication management despite differences in evaluations. However, our data suggest that children who received comprehensive evaluations had greater identification of comorbid conditions that may influence academic, behavioral, and social outcomes.


[Application of percutaneous pie-crusting deep medial collateral ligament release for posterior horn surgery of medial meniscus].
To explore clinical and radiographic effects of percutaneous pie-crusting deep medial collateral ligament release in patients with posterior horn tear of medial meniscus combined with tight medial compartment. From January 2012 to December 2016, 35 patients with medial meniscus posterior horn injury were treated with percutaneous pie crusting deep medial collateral ligament release technique, including 21 males and 14 females, aged from 21 to 55 years old with an average of (39.1±6.5) years old. Degree of meniscus extrusion were recorded before and 24 months after operation. The knee valgus stress test was performed to evaluate stability of medial collateral ligament, and compared difference between healthy and affected side. Lysholm and IKDC functional scores were compared before and 24 months after operation. All patients were followed up from 27 to 60 months with an average of (36.7±6.8) months. All patients were underwent operation, the wound healed well without complications. Operative time ranged from 0.5 to 1.2 h with an average of (0.8±0.4) h. Nineteen patients were performed partial meniscectomy, 16 patients were performed repair suture. Convex of meniscus before operation was (1.5±0.7) mm, and (1.7±0.4) mm after operation;had no statistical difference(P>0.05). Lysholm score was improved from 53.4±8.8 before operation to 91.5±4.6 at 24 months after operation;IKDC score was increased from 50.7±9.2 before operation to 90.6±3.9 at 24 months after operation;there was statistically significant (P<0.05). Valgus stress test was performed on 0 ° and 30 ° position of knee flexion in affected side and compared with ipsilateral side, all patients showed negative. For patients with medial meniscus tear of posterior horn combined with tight medial compartment, percutaneous pie-crusting deep medial collateralligament release could improve medial compartment space, and Knee valgus instability and meniscus extrusion are not affected.


Synthesis and Morphological Control of Biocompatible Fluorescent/Magnetic Janus Nanoparticles Based on the Self-Assembly of Fluorescent Polyurethane and Fe₃O₄ Nanoparticles.
Functionalized Janus nanoparticles have received increasing interest due to their anisotropic shape and the particular utility in biomedicine areas. In this work, a simple and efficient method was developed to prepare fluorescent/magnetic composite Janus nanoparticles constituted of fluorescent polyurethane and hydrophobic nano Fe₃O₄. Two kinds of fluorescent polyurethane prepolymers were synthesized by the copolymerization of fluorescent dye monomers, and the fluorescent/magnetic nanoparticles were fabricated in one-pot via the process of mini-emulsification and self-assembly. The nanostructures of the resulting composite nanoparticles, including core/shell and Janus structure, could be controlled by the phase separation in assembly process according to the result of transmission electron microscopy, whereas the amount of the nonpolar segments of polyurethane played an important role in the particle morphology. The prominent magnetic and fluorescent properties of the Janus nanoparticles were also confirmed by vibrating magnetometer and confocal laser scanning microscope. Furthermore, the Janus nanoparticles featured excellent dispersity, storage stability, and cytocompatibility, which might benefit their potential application in biomedical areas.


Oxidized high-density lipoprotein impairs endothelial progenitor cells' function by activation of CD36-MAPK-TSP-1 pathways.
High-density lipoprotein (HDL) levels inversely correlate with cardiovascular events due to the protective effects on vascular wall and stem cells, which are susceptible to oxidative modifications and then lead to potential pro-atherosclerotic effects. We proposed that oxidized HDL (ox-HDL) might lead to endothelial progenitor cells (EPCs) dysfunction and investigated underlying mechanisms. ox-HDL was shown to increase apoptosis and intracellular reactive oxygen species levels, but to reduce migration, angiogenesis, and cholesterol efflux of EPCs in a dose-dependent manner. p38 mitogen-activated protein kinase (MAPK) and NF-κB were activated after ox-HDL stimulation, which also upregulated thrombospondin-1 (TSP-1) expression without affecting vascular endothelial growth factor. Effects caused by ox-HDL could be significantly attenuated by pretreatment with short hairpin RNA-mediated CD36 knockdown or probucol. Data of in vivo experiments and the inverse correlation of ox-HDL and circulating EPC numbers among patients with coronary artery diseases (CAD) or CAD and type 2 diabetes also supported it. Meanwhile, HDL separated from such patients could significantly increase cultured EPC's caspase 3 activity, further supporting our proposal. This is the most complete study to date of how ox-HDL would impair EPCs function, which was involved with activation of CD36-p38 MAPK-TSP-1 pathways and proved by not only the inverse relationship between ox-HDL and circulating EPCs in clinic but also pro-apoptotic effects of HDL separated from patients' serum. Activation of CD36-p38 MAPK-TSP-1 pathways contributes to the pathological effects of ox-HDL on EPCs' dysfunction, which might be one of the potential etiological factors responsible for the disturbed neovascularization in chronic ischemic disease.


Prooxidative chain transfer activity by thiol groups in biological systems.
Cysteine is arguably the best-studied biological amino acid, whose thiol group frequently participates in catalysis or ligand binding by proteins. Still, cysteine's unusual biological distribution has remained mysterious, being strikingly underrepresented in transmembrane domains and on accessible protein surfaces, particularly in aerobic life forms ("cysteine anomaly"). Noting that lipophilic thiols have been used for decades as radical chain transfer agents in polymer chemistry, we speculated that the rapid formation of thiyl radicals in hydrophobic phases might provide a rationale for the cysteine anomaly. Hence, we have investigated the effects of dodecylthiol and related compounds in isolated biomembranes, cultivated human cells and whole animals (C. elegans). We have found that lipophilic thiols at micromolar concentrations were efficient accelerators, but not inducers of lipid peroxidation, catalyzed fatty acid isomerization to trans-fatty acids, and evoked a massive cellular stress response related to protein and DNA damage. These effects were specific for lipophilic thiols and were absent with thioethers, alcohols or hydrophilic compounds. Catalytic chain transfer activity by thiyl radicals appears to have deeply influenced the structural biology of life as reflected in the cysteine anomaly. Chain transfer agents represent a novel class of biological cytotoxins that selectively accelerate oxidative damage in vivo.


CircSmg5 stimulates the osteogenic differentiation of bone marrow mesenchymal stem cells by targeting the miR-194-5p/Fzd6 axis and beta-catenin signaling.
Osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) is closely associated with bone diseases. Circular RNAs are reported to be involved in BMSC differentiation. CircSmg5 (circ_0001145) has been identified to be downregulated in an osteoporosis mouse model. In this study, we aimed to explore the function and regulatory mechanism of circSmg5 in BMSC osteogenic differentiation. The Alizarin Red staining and alkaline phosphatase staining assays were performed to explore the osteogenic differentiation of BMSCs. The interaction between circ_0001145, miR-194-5p, and frizzled class receptor 6 (Fzd6) was analyzed by luciferase reporter assay. The nuclear translocation of β-catenin was assessed using immunofluorescence staining. CircSmg5 is in stable circular structure. CircSmg5 expression was elevated in the process of BMSC osteogenic differentiation. CircSmg5 overexpression promoted the osteogenic differentiation of BMSCs. CircSmg5 bound with miR-194-5p, whose expression was decreased in the osteogenic differentiation of BMSCs. MiR-194-5p directly targeted the 3'UTR of Fzd6. The mRNA and protein levels of Fzd6 were positively modulated by circSmg5 and negatively regulated by miR-194-5p in BMSCs. CircSmg5 was demonstrated to promote the BMSC osteogenic differentiation by targeting the miR-194-5p/Fzd6 axis to activate the Wnt/β-catenin signaling.


Toll-like Receptor as a Molecular Link between Metabolic Syndrome and Inflammation: A Review.
Metabolic Syndrome (MetS) involves a cluster of five conditions, i.e. obesity, hyperglycaemia, hypertension, hypertriglyceridemia and low High-Density Lipoprotein (HDL) cholesterol. All components of MetS share an underlying chronic inflammatory aetiology, manifested by increased levels of pro-inflammatory cytokines. The pathogenic role of inflammation in the development of MetS suggested that toll-like receptor (TLR) activation may trigger MetS. This review summarises the supporting evidence on the interactions between MetS and TLR activation, bridged by the elevation of TLR ligands during MetS. The regulatory circuits mediated by TLR activation, which modulates signal propagation, leading to the state of chronic inflammation, are also discussed. Taken together, TLR activation could be the molecular basis in the development of MetS-induced inflammation.


Mapping genes for phosphorus utilization and correlated traits using a 4k SNP linkage map in Japanese quail (Coturnix japonica).
A large F2 cross with 920 Japanese quail was used to map QTL for phosphorus utilization, calcium utilization, feed per gain and body weight gain. In addition, four bone ash traits were included, because it is known that they are genetically correlated with the focal trait of phosphorus utilization. Trait recording was done at the juvenile stage of the birds. The individuals were genotyped genome-wide for about 4k SNPs and a linkage map constructed, which agreed well with the reference genome. QTL linkage mapping was performed using multimarker regression analysis in a line cross model. Single marker association mapping was done within the mapped QTL regions. The results revealed several genome-wide significant QTL. For the focal trait phosphorus utilization, a QTL on chromosome CJA3 could be detected by linkage mapping, which was substantiated by the results of the SNP association mapping. Four candidate genes were identified for this QTL, which should be investigated in future functional studies. Some overlap of QTL regions for different traits was detected, which is in agreement with the corresponding genetic correlations. It seems that all traits investigated are polygenic in nature with some significant QTL and probably many other small-effect QTL that were not detectable in this study.


[Optimization of the functional activity of the adrenal cortex in patients with sepsis].
The effect of hemosorption on adrenal steroidogenic reactions has been studied in 75 patients with sepsis. Different variants of steroidogenesis optimization have been used. Hemosorption led to an increase in the level of steroid hormones, with this background retained throughout the whole treatment period. The most effective of all the variants of stress adrenal steroidogenic reactions optimization was the application of B, B1, B6, B12 vitamin complex and taktivin. The least effective was the application of B vitamin complex, taktivin and ultraviolet blood irradiation. The technique elaborated makes it possible to avoid an increase in steroid blood level during treatment, which must have a favorable effect on the recovery of immune homeostasis in the patients.


Descriptive observational study on early functional recovery and postoperative pain in knee joint replacement surgery.
The management of postoperative pain in joint replacement surgery represents a challenge. Therefore, the advent of new strategies in the management of pain such as local infiltration analgesia (LIA) and the adductor canal block, allowed an adequate postoperative pain control and early rehabilitation of the patient in total knee arthroplasty. A descriptive, observational and longitudinal study of a prospective cohort of patients operated by the group of joint replacements between September 29, 2017 and November 30, 2017 was conducted. They were evaluated with the analog pain scale for postoperative pain at 5 different times (1:Upon leaving recovery; 2:On the first postoperative day; 3:Before initiating in-hospital physical therapy; 4:At the end of physical therapy, and 5:Upon departure from the clinic). Of the 141 patients operated on, 70.9% of the patients were managed with LIA and 29.1% with adductor canal block. There were no differences between groups in postoperative pain assessment during the 5 different times (P≥.45). Our results indicate that LIA and the adductor canal blockade demonstrated the same efficacy for the control of postoperative pain and rehabilitation.


Revisiting the factorial validity of the Revised UCLA Loneliness Scale: a test of competing models in a sample of teachers.
Over the years, various studies addressing different populations have consistently raised concerns about the unidimensionality of the Revised UCLA Loneliness Scale. In the present study, the factorial structure of the scale was examined by comparing it to alternative models. In a sample of 1,157 French Canadian teachers, results of confirmatory factor analysis support a three-factor model solution. Support for the invariance of this model across sexes and teaching levels (i.e., elementary and high school) was also obtained.


Modulation of internal model formation during force field-induced motor learning by anodal transcranial direct current stimulation of primary motor cortex.
Human subjects can quickly adapt and maintain performance of arm reaching when experiencing novel physical environments such as robot-induced velocity-dependent force fields. Using anodal transcranial direct current stimulation (tDCS) this study showed that the primary motor cortex may play a role in motor adaptation of this sort. Subjects performed arm reaching movement trials in three phases: in a null force field (baseline), in a velocity-dependent force field (adaptation; 25 N s m(-1)) and once again in a null force field (de-adaptation). Active or sham tDCS was directed to the motor cortex representation of biceps brachii muscle during the adaptation phase of the motor learning protocol. During the adaptation phase, the global error in arm reaching (summed error from an ideal trajectory) was similar in both tDCS conditions. However, active tDCS induced a significantly greater global reaching (overshoot) error during the early stage of de-adaptation compared to the sham tDCS condition. The overshoot error may be representative of the development of a greater predictive movement to overcome the expected imposed force. An estimate of the predictive, initial movement trajectory (signed error in the first 150 ms of movement) was significantly augmented during the adaptation phase with active tDCS compared to sham tDCS. Furthermore, this increase was linearly related to the change of the overshoot summed error in the de-adaptation process. Together the results suggest that anodal tDCS augments the development of an internal model of the novel adapted movement and suggests that the primary motor cortex is involved in adaptation of reaching movements of healthy human subjects.


The development of paediatric surgery in Papua New Guinea.
Until 1993 complex surgery for children in Papua New Guinea (PNG) was usually conducted by general surgeons, or by overseas paediatric surgeons during ad hoc visits. There had been little coordination of these occasional international visits and no involvement in the surgical teaching program in the University Department of Surgery. Over eight years from July 1993 to July 2000, three Australian surgeons spent 33 weeks teaching paediatric surgery and paediatric urology, using lectures, tutorials and operative demonstration sessions. This paper is a report of the teaching and service provided by one surgeon (PAD) over 27 weeks in PNG and describes the development and training of the surgeon who will be the country's first qualified paediatric surgeon (MM). Other support given has been assistance with the publication of scientific papers, reviews of Masters' theses, the development of a proposal for investigation of the prevalence of renal tract anomalies and the development of protocols for the surgical management of anorectal anomalies and Hirschsprung's disease. In addition to this teaching, 311 children have had surgery during the 11 visits that form the basis of this report. The visits have been supported and funded by AusAID and the Royal Australasian College of Surgeons through the Medical Officer, Nursing and Allied Health Professional (MONAHP) and Pacific Islands Project (PIP) programs. An indication of the impact on the care of children with surgical diseases is evident from the improved skills and the changed referral patterns over the eight years.


Evaluation of Bacillus thuringiensis Subsp. Israelensis and Bacillus sphaericus Combination Against Culex pipiens in Highly Vegetated Ditches.
Among the few mosquito larvicides available in the market, Bacillus thuringiensis subsp. israelensis (Bti) and B. sphaericus (Bs) represent the most environmentally safe alternatives. The combination of the 2 products is known to overcome their specific limitations by producing a synergistic effect. The aim of the study was to assess the effect and persistence of a single treatment with a granular Bti + Bs formulation on highly vegetated ditches in northeastern Italy that represents the primary rural larval sites for Culex pipiens, the primary vector of the West Nile virus in Europe. The analysis takes into account the nonlinear temporal effects on the population dynamics of larvae and pupae. The results showed a dramatic reduction in mosquito larval abundance 24 h posttreatment (93%) and was effective against larvae up to 22 days (100%). The residual effect after 28 days was 99.5%, and a limited residual effect was observed after 39 days (31.2%). A reduction in pupal density was observed after 4 days (70%) and was >98% from days 14 to 28 posttreatment, persisting for up to 39 days (84% after 39 days). The results demonstrate the effective use of the Bti + Bs formulation against Cx. pipiens in vegetated ditches in rural areas. Our modeling framework provides a flexible statistical approach to predict the residual effect of the product over time, in order to plan a seasonal intervention scheme.


Active and Former Injection Drug Users Report of HIV Risk Behaviors During Periods of Incarceration.
American prisons have increasing numbers of inmates incarcerated for drug offenses. This population is at high risk for HIV-infection and may continue HIV transmission risk behaviors while incarcerated. We find that 31% of injection drug users with a history of imprisonment had used illicit drugs in prison, and nearly half of these persons had injected drugs while incarcerated. Male gender and number of times incarcerated were associated with drug use in prison. Interventions for drug-using prisoners that are advocated in some European prisons, such as needle exchange programs and methadone maintenance, need attention in the United States.


Sudden death in a traveler following halofantrine administration--Togo, 2000.
On July 17, 2000, a previously healthy 22-year-old U.S. student collapsed and died suddenly while leading a teenage exchange group in West Africa. This report summarizes the results of the investigations of this incident, which implicate use of halofantrine for treatment of malaria as the cause of death. Travelers should be warned that halofantrine treatment may be dangerous in persons with cardiac abnormalities or in those taking mefloquine for malaria prophylaxis.


Custom-made trays for targeted treatment of oral vesiculobullous diseases: A proposed protocol.
Topical and intralesional corticosteroids (CS) are effective treatments for oral manifestations of autoimmune vesiculobullous diseases (VBD); however, may cause unnecessary absorption in the adjacent healthy mucosal surfaces, local infections, atrophy, and injection pain. To propose a protocol using a unique modality for targeted administration of CS for the treatment of local manifestations of oral VBD. The medical records of nine patients with systemic CS-resistant VBD oral lesions, were reviewed following 3 weeks of treatment and 3 months of tapering using a custom-made tray designed for precise application of topical CS targeted to the involved mucosa. Following treatment, lesions have healed in 8/9 patients. Involvement of surrounding mucosa was minimal, and no systemic adverse effects were recorded. The proposed protocol using custom-made trays for targeted CS treatment could be an effective alternative treatment for oral VBD.


Effects of commonly used food additives on haematological parameters of Wistar rats.
This study was done to investigate the effects of common food additives such as sodium benzoate (SB) and ascorbic acid (AA) on haematological parameters of male Wistar rats. Forty-eight (48) male albino rats with an average weight of 105 g were grouped into twelve (n = 4) of Basal Control and other 11 groups orally administered 1 mg of SB, 10 mg of SB, 10 mg of AA, 0.2 mg of AA + 0.5 mg of SB, 0.2 mg of AA + 1 mg of SB, 0.2 mg of AA + 10 mg of SB, 0.2 mg of SB + 0.1 mg of AA, 0.2 mg of SB + 0.5 mg of AA, carbonated soft drinks (CSD)+ 0.1 mg of AA, CSD + 1 mg of AA and CSD + 10 mg of AA, respectively for 21 non-consecutive days. At the end of the experiment, blood samples were collected in EDTA anticoagulant tubes, haematological parameters were evaluated, and data were analyzed. There was a dose-dependent increase (p < 0.05) in White Blood Cell counts of SB treated rats compared with the control group. The lymphocyte exhibited significant reduction (p < 0.05) in the groups treated with 1mg SB and 10mg SB/kg bodyweight of 67 ± 2.96 and 58 ± 4.18%, respectively. The mean corpuscular haemoglobin showed no significant difference at 95% confidence interval. However, mean corpuscular haemoglobin concentration, haematocrit and platelet were affected by an increase in the concentrations of SB. High SB concentrations increased the destruction of erythrocytes, which directly increased the catabolism of haemoglobin. However, AA administration mitigated the adverse effects of SB on the haematological parameters of the animal.


P53 gene mutations in acute myeloid leukemia with 17p monosomy.
We looked for mutations of exons 5 to 8 of the P53 gene in 10 patients with acute myeloid leukemia (AML) and 17p monosomy, and 36 patients with AML and no cytogenetic abnormalities of 17p. DNA was analyzed by polymerase chain reaction, single-strand conformation polymorphism analysis, and nucleotide sequencing. Four of the 10 patients with 17p monosomy showed point mutation, single-nucleotide deletion, or insertion in exons 7 or 8. By contrast, only 1 of the 36 patients with AML and no cytogenetic abnormalities of 17p showed a mutation of the P53 gene in exons 5 to 8 (P less than .01). These results suggest that alterations of the P53 gene may have a role in leukemogenesis in some cases of AML. The fact that P53 gene mutations occurred more often in patients with 17p monosomy seems to support the "recessive" model of tumor suppressive activity of the P53 gene rather than the "dominant" model, in which alteration of only one allele is sufficient for the development of malignancy.


[Chemotherapy-induced hemolytic uremic syndrome. A complication to mitomycin treatment of breast carcinoma].
Mitomycin is a cytotoxic antibiotic that was first introduced into clinical use in 1958. Not until twenty years later was it recognised that haemolytic uremic syndrome could develop after treatment with mitomycin. It can be asked whether this condition was undiagnosed in previous years, since its frequency is now reported to be 4-15%. The disease appears to be dose-related, since it rarely occurs in patients who have received mitomycin < 30 mg/m2. No effective therapy has been established. We describe two patients with breast cancer in remission after treatment with mitomycin in combination with 5-fluorouracil. Both developed haemolytic uremic syndrome with fatal outcome.


Unbalanced MOF-on-MOF growth for the production of a lopsided core-shell of MIL-88B@MIL-88A with mismatched cell parameters.
The unbalanced MOF-on-MOF growth of MIL-88A on the MIL-88B template, where both MOFs have a similar three-dimensional hexagonal structure but with mismatched cell parameters, results in the formation of an atypical lopsided core-shell of MIL-88B@MIL-88A with an off-centered core. The formation mechanism of the lopsided core-shell of MIL-88B@MIL-88A is verified via monitoring the growth process.


Contingency management for smoking cessation in adolescent smokers.
This pilot study evaluated the use of contingency management (CM) procedures in combination with cognitive-behavioral therapy (CBT) for smoking cessation in adolescents. Twenty-eight treatment-seeking adolescent smokers participated in a 1-month, school-based smoking cessation program and were randomly assigned to receive either CM with weekly CBT or CBT alone. In the CM+CBT group, biochemical verification of abstinence was obtained twice daily during the first 2 weeks, followed by daily appointments during the 3rd week and once every other day during the 4th week. Participants were monetarily reinforced for abstinence on an escalating magnitude schedule with a reset contingency. At the end of 1 week and 1 month of treatment, abstinence verified using quantitative urine cotinine levels was higher in participants in the CM+CBT group (1 week: 76.7%; 1 month: 53.0%) when compared with the CBT-alone group (1 week: 7.2%; 4 weeks: 0%). These preliminary results provide a strong initial signal supporting the utility of CM techniques for smoking cessation in adolescents and demonstrate the feasibility of implementing such a program in a school setting.


Theoretical study of unimolecular decomposition of catechol.
This study develops the reaction pathway map for the unimolecular decomposition of catechol, a model compound for various structural entities present in biomass, coal, and wood. Reaction rate constants at the high-pressure limit are calculated for the various possible initiation channels. It is found that catechol decomposition is initiated dominantly via hydroxyl H migration to a neighboring ortho carbon bearing an H atom. We identify the direct formation of o-benzoquinone to be unimportant at all temperatures, consistent with the absence of this species from experimental measurements. At temperatures higher than 1000 K, water elimination through concerted expulsion of a hydroxyl OH together with an ortho H becomes the most significant channel. Rice-Ramsperger-Kassel-Marcus simulations are performed to establish the branching ratio between these two important channels as a function of temperature and pressure. All unimolecular routes to the reported major experimental products (CO, 1,3-C(4)H(6) and cyclo-C(5)H(6)) are shown to incur large activation barriers. The results presented herein should be instrumental in gaining a better understanding of the decomposition behavior of catechol-related compounds.


Bayesian Approach to Multivariate Component-Based Logistic Regression: Analyzing Correlated Multivariate Ordinal Data.
Applications of component-based models have gained much attention as a means of accompanying dimension reduction in the regression setting and have been successfully implemented to model a univariate outcome in the behavioral and social sciences. Despite the prevalence of correlated ordinal outcome data in the fields, however, most of the extant component-based models have been extended to address the multivariate ordinal issue with a simplified but unrealistic assumption of independence, which may lead to biased statistical inferences. Thus, we propose a Bayesian methodology for a component-based model that accounts for unstructured residual covariances, while regressing multivariate ordinal outcomes on pre-defined sets of predictors. The proposed Bayesian multivariate ordinal logistic model re-expresses ordinal outcomes of interest with a set of latent continuous variables based on an approximate multivariate t-distribution. This contributes not only to developing an efficient Gibbs sampler, a Markov Chain Monte Carlo algorithm, but also to facilitating the interpretation of regression coefficients as log-transformed odds ratio. The empirical utility of the proposed method is demonstrated through analyzing a subset of data, extracted from the 2009 to 2010 Health Behavior in School-Aged Children study that investigates risk factors of four different forms of bullying perpetration and victimization: physical, social, racial, and cyber.


Human cytomegalovirus activation of ERK and myeloid cell leukemia-1 protein correlates with survival of latently infected cells.
The ability of human CMV (HCMV) to enter and establish a latent infection in myeloid cells is crucial for survival and transmission in the human population. Initial pathogen binding and entry triggers a number of antiviral responses, including the activation of proapoptotic cell death pathways, which must be countered during latency establishment. However, mechanisms responsible for a prosurvival state in myeloid cells upon latent HCMV infection remain completely undefined. We hypothesized that the cellular antiapoptotic machinery must be initially activated by HCMV to promote early survival events upon entry. Here we show that HCMV transiently protects nonpermissive myeloid cells from chemical and virus entry induced cell death by up-regulating a key myeloid cell survival gene, myeloid cell leukemia (MCL)-1 protein. The induction of MCL-1 expression was independent of viral gene expression but dependent on activation of the ERK-MAPK pathway by viral glycoprotein B. Inhibition of ERK-MAPK signaling, inhibition of HCMV fusion, antibody-mediated neutralization of glycoprotein B signaling or expression of a shRNA against MCL-1 all correlated with increased cell death in response to virus infection or chemical stimulation. Finally we show that activation of ERK-MAPK signaling impacts on long-term latency and reactivation in hematopoietic cells. Thus, HCMV primes myeloid cells for from the initial virus-cell encounter. Given the importance of ERK and MCL-1 for myeloid cell survival, the successful establishment of HCMV latency in myeloid progenitors begins at the point of virus entry.


Capsicum annuum L. methanolic extract inhibits ovalbumin-induced airway inflammation and oxidative stress in a mouse model of asthma.
The pepper fruit of Capsicum annuum L. is used as a food, spice, and topical medicine. Here, we investigated the effect of a methanolic C. annuum L. extract (CAE) in a mouse model of ovalbumin-induced allergic airway inflammation. Animals were treated with CAE by oral gavage before ovalbumin challenge. After ovalbumin challenge, airway responsiveness to methacholine, influx of inflammatory cells into the lung, cytokine levels in bronchoalveolar lavage fluid and lung, nuclear factor-κB (NF-κB) activity in lungs, and lung histopathology were assessed. Oral treatment with CAE significantly reduced the pathophysiological signs of allergic airway disease, including increased inflammatory cell recruitment to the airways, airway hyperresponsiveness, and increased levels of T-helper type 2 cytokines. Reactive oxygen species were also decreased in cells from bronchoalveolar lavage fluid. In addition, we found that administration of CAE attenuated ovalbumin-induced increases in NF-κB activity in lungs. Collectively, these results suggest that CAE may be an effective oral treatment for allergic airway inflammation by virtue of its antioxidant activity.


Activation of store-operated channels by noradrenaline via protein kinase C in rabbit portal vein myocytes.
In the present study we have investigated the role of diacylglycerol (DAG) and protein kinase C (PKC) in mediating activation of Ca(2+)-permeable store-operated channels (SOCs) by noradrenaline in rabbit portal vein smooth muscle cells. With cell-attached recording, bath application of noradrenaline, 1-oleoyl-acetyl-sn-glycerol (OAG) and phorbol 12,13-dibutyrate (PDBu) evoked single channel currents. The biophysical properties of these channel currents were similar to those of the channel currents activated by depletion of internal Ca(2+) stores with cyclopiazonic acid (CPA). The activation of SOCs in cell-attached recording by noradrenaline, OAG, PDBu, CPA and the acetoxymethyl ester form of BAPTA (BAPTA-AM) was markedly inhibited by the PKC inhibitors chelerythrine and RO-31-8220. In isolated outside-out patches CPA did not evoke SOCs but noradrenaline stimulated SOC activity, which was reduced by about 90 % by PKC inhibitors. The addition of the serine/threonine phosphatase inhibitors calyculin A and microcystin also stimulated SOCs in isolated outside-out patches. It is concluded that in rabbit portal vein myocytes, noradrenaline activates SOCs via DAG and PKC, possibly by a store-independent mechanism. In addition in this cell type it appears that PKC and phosphorylation may play an important role in stimulating SOC activity in response to depletion of internal Ca(2+) stores by CPA and BAPTA-AM.


Non-communicable respiratory disease in Malawi: a systematic review and meta-analysis.
Non-communicable respiratory diseases are important contributors to morbidity and mortality in sub-Saharan African countries such as Malawi. To conduct a systematic review of the available literature relating to chronic respiratory disease in Malawi. We conducted a systematic protocol-driven literature search of key scientific databases including Scopus and Medline. Papers were independently assessed for eligibility by two authors and included if they reported objective measures (including self-reported standard symptoms) of chronic respiratory disease and were conducted in Malawi. A meta-analysis of available estimates was then conducted. We re-analysed data from three of these studies in a secondary data analysis to allow for between-study comparisons. Our search identified 393 papers of which 17 (5 involving children and 12 involving adults) met the inclusion criteria. Wheeze was the symptom most frequently reported in children in the community (12.1%), hospital (11.2%) and HIV clinic (8.1%) settings. Cough was the symptom most frequently reported by adults in the community (3-18%). Spirometric abnormalities varied substantially between studies. For example, in adults, airflow obstruction varied between 2.3% and 20% and low forced vital capacity (FVC) varied between 2.7% and 52.8%. We identified a high burden of chronic respiratory symptoms and abnormal spirometry (particularly low FVC) within paediatric and adult populations in Malawi. The estimates for country-wide burden related to this disease were limited by the heterogeneity of the methods used to assess symptoms and spirometry. There is an urgent need to develop a better understanding of the determinants and natural history of non-communicable respiratory disease across the life-course in Malawi.


Issues and challenges facing rural hospitals.
Australia's rural hospitals face most of the same issues and challenges faced by metropolitan hospitals. However they also face additional challenges around geographic isolation, their iconic status and role as major employers in local communities, and their close relationship with community based health services. Future opportunities include more formal integration with community service through multipurpose service arrangements, regional networking with urban centers, expanded us of IT linkages, and with the expanding rural academic networks. Rural hospitals' roles as key aged care providers in the country is a particular challenge.


Cost-Effectiveness of Fecal Microbiota Transplantation in the Treatment of Recurrent Clostridium Difficile Infection: A Literature Review.
Clostridium difficile (C. difficile) is a common cause of antibiotic--associated diarrhea (AAD), being responsible for 15--25% of all AAD cases. The purpose of this literature review is to determine the cost-effectiveness of fecal microbiota transplantation (FMT) and how it compares in this regard to the standard treatments of choice for recurrent C. difficile infection (CDI). The review of the literature along with the evaluation of three comparative cost effective analyses yielded findings consistent with the view that FMT is the most cost-effective option in treating recurrent CDI. There are some (but considerably less) data indicating that FMT may be a cost effective strategy in treating initial CDI, as well. The superior cost-effectiveness of FMT as compared to the preferred standards of treatment for recurrent CDI suggest FMT use should become more integrated in routine clinical practice. Increased utilization of FMTs would allow for better control of this increasingly problematic disease as well as lower costs associated with its management.


Low density lipoproteins develop resistance to oxidative modification due to inhibition of cholesteryl ester transfer protein by a monoclonal antibody.
Although numerous studies have investigated the relationship between cholesteryl ester transfer protein (CETP) and high density lipoprotein (HDL) remodeling, the relationship between CETP and low density lipoproteins (LDL) is still not fully understood. In the present study, we examined the effect of the inhibition of CETP on both LDL oxidation and the uptake of the oxidized LDL, which were made from LDL under condition of CETP inhibition, by macrophages using a monoclonal antibody (mAb) to CETP in incubated plasma. The 6-h incubation of plasma derived from healthy, fasting human subjects led to the transfer of cholesteryl ester (CE) from HDL to VLDL and LDL, and of triglycerides (TG) from VLDL to HDL and LDL. These net mass transfers of neutral lipids among the lipoproteins were eliminated by the mAb. The incubation of plasma either with or without the mAb did not affect the phospholipid compositions in any lipoproteins. As a result, the LDL fractionated from the plasma incubated with the mAb contained significantly less CE and TG in comparison to the LDL fractionated from the plasma incubated without the mAb. The percentage of fatty acid composition of LDL did not differ among the unincubated plasma, the plasma incubated with the mAb, and that incubated without the mAb. When LDL were oxidized with CuSO4, the LDL fractionated from the plasma incubated with the mAb were significantly resistant to the oxidative modification determined by measuring the amount of TBARS and by continuously monitoring the formation of the conjugated dienes, in comparison to the LDL fractionated from the plasma incubated without the mAb. The accumulation of cholesteryl ester of oxidized LDL, which had been oxidized for 2 h with CuSO4, in J774.1 cells also decreased significantly in the LDL fractionated from the plasma incubated with mAb in comparison to the LDL fractionated from the plasma incubated without the mAb. These results indicate that CETP inhibition reduces the composition of CE and TG in LDL and makes the LDL resistant to oxidation. In addition, the uptake of the oxidized LDL, which was made from the LDL under condition of CETP inhibition, by macrophages also decreased.


A globular protein with slower amide proton exchange from an alpha helix than from antiparallel beta sheets.
In proteinase inhibitor IIA from bull seminal plasma, which is a small globular protein with 57 amino acid residues, measurements of individual amide proton exchange rates by two-dimensional correlated 1H NMR spectroscopy (COSY) showed that the exchange was slowest for some hydrogen bonded amide groups in an alpha-helix. This contrasts with all other proteins which were so far studied in detail, where the slowest exchange rates were observed for hydrogen bonded amide protons in antiparallel beta-sheets.


NRP-1 Receptor Expression Mismatch in Skin of Subjects with Experimental and Diabetic Small Fiber Neuropathy.
The in vivo cutaneous nerve regeneration model using capsaicin is applied extensively to study the regenerative mechanisms and therapeutic efficacy of disease modifying molecules for small fiber neuropathy (SFN). Since mismatches between functional and morphological nerve fiber recovery are described for this model, we aimed at determining the capability of the capsaicin model to truly mimic the morphological manifestations of SFN in diabetes. As nerve and blood vessel growth and regenerative capacities are defective in diabetes, we focused on studying the key regulator of these processes, the neuropilin-1 (NRP-1)/semaphorin pathway. This led us to the evaluation of NRP-1 receptor expression in epidermis and dermis of subjects presenting experimentally induced small fiber neuropathy, diabetic polyneuropathy and of diabetic subjects without clinical signs of small fiber neuropathy. The NRP-1 receptor was co-stained with CD31 vessel-marker using immunofluorescence and analyzed with Definiens® technology. This study indicates that capsaicin application results in significant loss of epidermal NRP-1 receptor expression, whereas diabetic subjects presenting small fiber neuropathy show full epidermal NRP-1 expression in contrast to the basal expression pattern seen in healthy controls. Capsaicin induced a decrease in dermal non-vascular NRP-1 receptor expression which did not appear in diabetic polyneuropathy. We can conclude that the capsaicin model does not mimic diabetic neuropathy related changes for cutaneous NRP-1 receptor expression. In addition, our data suggest that NRP-1 might play an important role in epidermal nerve fiber loss and/or defective regeneration and that NRP-1 receptor could change the epidermal environment to a nerve fiber repellant bed possibly through Sem3A in diabetes.


2D:4D Ratio Differs in Ischemic Stroke: A Single Center Experience.
The index to ring finger length (2D:4D) ratio is a proxy biomarker for prenatal exposure of sex hormones. Sex hormones are associated with the pathogenesis of ischemic stroke. The purpose of the study was to demonstrate the association between 2D:4D and ischemic stroke. This study retrospectively reviewed the data of 100 patients with first ever ischemic stroke between September, 2016 and June, 2017. The lengths of index finger and ring finger of both hands were measured using electronic calipers and calculated for 2D:4D ratios. Receive operating characteristic (ROC) mode was used to detect predicting performance of 2D:4D ratios for ischemic stroke. 2D:4D ratios in ischemic stroke patients were higher than controls in both hands (P < 0.05), except right 2D:4D ratio in females. The ROC analyses showed that the area under the curve (AUC) were 0.635 (95%CI: 0.527-0.743) for left 2D:4D ratio, and 0.647 for right (95%CI: 0.539-0.755) (P < 0.05). The AUC of left and right 2D:4D ratio in male were 0.667 (95%CI: 0.514-0.820) and 0.670 (95%CI: 0.519-0.822) (P < 0.05). In female, no significance were found in ROC analysis. And there were no correlation between 2D:4D value and stroke severity (P > 0.05). The current study indicated that the diagnostic value of 2D:4D ratio was limited in ischemic stroke. Further research is required to explore the role of it in screening ischemic stroke.


Unraveling algal lipid metabolism: Recent advances in gene identification.
Microalgae are now the focus of intensive research due to their potential as a renewable feedstock for biodiesel. This research requires a thorough understanding of the biochemistry and genetics of these organisms' lipid-biosynthesis pathways. Genes encoding lipid-biosynthesis enzymes can now be identified in the genomes of various eukaryotic microalgae. However, an examination of the predicted proteins at the biochemical and molecular levels is mandatory to verify their function. The essential molecular and genetic tools are now available for a comprehensive characterization of genes coding for enzymes of the lipid-biosynthesis pathways in some algal species. This review mainly summarizes the novel information emerging from recently obtained algal gene identification.


Cloning and enhanced expression of the cytochrome P450nor gene (nicA; CYP55A5) encoding nitric oxide reductase from Aspergillus oryzae.
We cloned and characterized the gene and cDNA of Aspergillus oryzae cytochrome P450nor (Anor). The Anor gene (nicA; CYP55A5) has a different gene structure from other P450nor genes in that it has an extra intron. There were not only two kinds of mRNA but also two sets of TATA-box and CCAAT-box, and it appears that this gene has two expression patterns, like CYP55A1 of Fusarium oxysporum. A reporter analysis using the uidA gene indicated that gene expression of CYP55A5 was induced under anaerobic conditions, like CYP55A1. When the CYP55A5 gene was overexpressed in A. oryzae, a large amount of active Anor were accumulated as intracellular protein. Anor employed both NADH and NADPH as electron donors for reducing nitric oxide to nitrous oxide. Anor measured the amount of NO generated from 3-(2-Hydroxy-1-(1-methylethyl)-2-nitrosohydrazino)-1-propanamine (NOC5) with a spectrophotometer. The sensitivity was 10 nmol/ml.


A Container-Attachable Inertial Sensor for Real-Time Hydration Tracking.
Various sensors have been proposed to address the negative health ramifications of inadequate fluid consumption. Amongst these solutions, motion-based sensors estimate fluid intake using the characteristics of drinking kinematics. This sensing approach is complicated due to the mutual influence of both the drink volume and the current fill level on the resulting motion pattern, along with differences in biomechanics across individuals. While motion-based strategies are a promising approach due to the proliferation of inertial sensors, previous studies have been characterized by limited accuracy and substantial variability in performance across subjects. This research seeks to address these limitations for a container-attachable triaxial accelerometer sensor. Drink volume is computed using support vector machine regression models with hand-engineered features describing the container's estimated inclination. Results are presented for a large-scale data collection consisting of 1908 drinks consumed from a refillable bottle by 84 individuals. Per-drink mean absolute percentage error is reduced by 11.05% versus previous state-of-the-art results for a single wrist-wearable inertial measurement unit (IMU) sensor assessed using a similar experimental protocol. Estimates of aggregate consumption are also improved versus previously reported results for an attachable sensor architecture. An alternative tracking approach using the fill level from which a drink is consumed is also explored herein. Fill level regression models are shown to exhibit improved accuracy and reduced inter-subject variability versus volume estimators. A technique for segmenting the entire drink motion sequence into transport and sip phases is also assessed, along with a multi-target framework for addressing the known interdependence of volume and fill level on the resulting drink motion signature.


Upregulation of monocyte urokinase plasminogen activator receptor during human endotoxemia.
The receptor for urokinase-type plasminogen activator (uPAR) (CD87) plays an important role in leukocyte adhesion and migration. To assess the effect of endotoxin on cellular uPAR, uPAR expression was determined on leukocytes by fluorescence-activated cell sorter analysis in seven healthy subjects following intravenous injection of endotoxin (lot G; 4 ng/kg). Endotoxin induced a transient increase in uPAR expression on monocytes, reaching a 92% +/- 46% increase over baseline expression after 6 h (P < 0.05). Endotoxin did not influence uPAR expression on granulocytes, while uPAR remained undetectable on lymphocytes. Endotoxin also increased soluble uPAR levels in plasma (P < 0.05). Stimulation of human whole blood with endotoxin or gram-positive stimuli in vitro also resulted in an upregulation of monocyte uPAR expression. Although tumor necrosis factor alpha (TNF) upregulated monocyte uPAR expression, anti-TNF did not influence the endotoxin-induced increase in monocyte uPAR expression. These data suggest that infectious stimuli may influence monocyte function in vivo by enhancing the expression of uPAR.


Evaluation of Direct Oral Anticoagulation Therapy in Heart and Lung Transplant Recipients.
Anticoagulation therapy is common in thoracic transplant recipients. Direct oral anticoagulants (DOACs) are alternatives to warfarin therapy, but characterization of their use in solid organ transplant is absent. The primary objective of this study was to describe a thoracic transplant patient population initiated on DOAC therapy. Secondary objectives were to assess adverse reactions, venous thromboembolism (VTE) recurrence, and drug-drug interactions during DOAC therapy. Single-center retrospective cohort study. A tertiary care medical center including inpatient hospitalization and outpatient transplant clinic visits. Thoracic transplant recipients who were initiated on DOACs between May 1, 2011, and March 1, 2015, at the University of Pittsburgh Medical Center were included. A total of 37 patients were included in the analysis. A majority of the patients were lung transplant recipients (86.4%) with a median age of 60.7 years. Twenty-eight patients had a history of VTE. The primary indication for DOAC initiation was VTE (86.5%). Rivaroxaban (78.4%) was the most commonly utilized agent. Dose reductions for major drug interactions (37.8%), renal insufficiency (10.8%), or both (8.1%) occurred within the study. Two patients had breakthrough VTE during DOAC therapy. Eight bleeding events were reported in the cohort, one of which was considered a major bleed. There was no difference in the incidence of bleeding in patients with drug-drug interactions and without drug-drug interactions during DOAC therapy (26.0% vs 7.1%, P = .154). Direct oral anticoagulant therapy was well tolerated by thoracic transplant recipients. Drug interactions and renal dose adjustments were common.


Identification of human calculi with time-of-flight secondary ion mass spectrometry.
Time-of-flight secondary ion mass spectrometry was used to study four human calculi and to compare the results with those from twelve commercially available urinary calculi minerals including three organic compounds (L-cystine, uric acid and sodium urate). Phase identification of calcium phosphate compounds was carried out by considering the relative ion abundances of [Ca(2)O](+) and [CaPO(2)](+). Deprotonated [M-H](-) and protonated [M+H](+) uric acid were detected and used for component recognition in pure uric acid and in the mixed samples of struvite, calcium oxalate and uric acid. Iodine related to the medical history of a patient was also detected.


Coordination by registered dieticians for nutritional and dietary support in disaster in Japan.
Yamada, a town of Iwate Prefecture in north-eastern Japan, was struck by the tsunami from the Great East Japan Earthquake. In Yamada, it was challenging to manage nutritional and diet support for food aid because these services were unavoidably drawn out for several months in evacuation shelters. In Japan, food aid in disasters is often provided, divided and distributed erratically due to poor efforts made with regards to dietary support from the perspective of nutrition. The need for nationally registered dieticians to coordinate nutritional and dietary support in evacuation shelters was considered in this disaster. A dietary support team was formed of nationally registered dieticians to study the dietary conditions of evacuees in shelters in Yamada and to develop a system to ensure the nutritional and balanced dietary needs of the evacuees. In this disaster response, model menus were prepared and a menu-food matching system was put in place to order and distribute foods required for balanced meals. Every effort was made to avoid excesses and deficiencies in nutrition; the meals consisted of a staple, main dish, side dish and soup. Along with that, food sanitation and stock management were improved. The menu-food matching system put together by the nationally registered dieticians was useful for nutritional and dietary support in this particular disaster. It is recommended that similar nutritional and dietary support coordinated by nationally registered dieticians be considered for disaster management plans where appropriate.


Relationship between mucosal TNF-α expression and Th1, Th17, Th22 and Treg responses in Helicobacter pylori infection.
Helicobacter pylori (H. pylori)-induced gastric inflammation in the gastric mucosa and significantly increases the risk of developing gastritis and peptic ulcer disease (PUD). The objective of this research is to determine the role of tumor necrosis factor-α (TNF-α) expression in the gastric mucosa of patients with H. pylori-associated gastritis and PUD compared to uninfected patients, and we determined the relation between TNF-α expression and Th1/Th17/Th22, and Treg cells. Fifty-five patients with H. pylori-associated gastritis, 47 patients with H. pylori-associated PUD, and 48 uninfected patients were in this research. Antrum biopsy was used to detect H. pylori, virulence factors and histopathological assessments. Expression of TNF-α in the infected group was significantly higher than the uninfected group. Also, cagA/oipA-positive infected patients induce significantly more TNF-α expression than do cagA/oipA-negative infected patients. Expression of TNF-α was significantly increased in the PUD group than the gastritis group. Notably, TNF-α expression had a significant positive correlation with the frequency of Th1/Th17/Th22 lymphocytes in the PUD group. These findings indicate the importance of increasing TNF-α with Th1, Th17, Th22 responses increase as an important risk factor for PUD in context of H. pylori infection.


Ionosphere-Constrained Single-Frequency PPP with an Android Smartphone and Assessment of GNSS Observations.
With the development of Global Navigation Satellite System (GNSS) and the opening of Application Programming Interface (API) of Android terminals, the positioning research of Android terminals has attracted the attention of GNSS community. In this paper, three static experiments were conducted to analyze the raw GNSS observations quality and positioning performances of the smartphones. For the two experimental smartphones, the numbers of visible satellites with dual-frequency signals are unstable and not enough for dual-frequency Precise Point Positioning (PPP) processing all through the day. Therefore, the ionosphere-constrained single-frequency PPP model was employed to improve the positioning with the smartphones, and its performance was evaluated and compared with those of the Single Point Positioning (SPP) and the traditional PPP models. The results show that horizontal positioning accuracies of the smartphones with the improved PPP model are better than 1 m, while those with the SPP and the traditional PPP models are about 2 m.


Clinical trials: what cancer patients need to know. The British Association of Cancer United Patients (BACUP).
In this report we highlight guidance issued by BACUP, the British Association of Cancer United Patients, on what cancer patients need to know about clinical trials. The information is to help nurses, GPs and other health professionals give patients the information they need to make an informed decision about participating in a trial.


Implementation of an enterprise risk-management program in a community teaching hospital.
As the complexity of healthcare and expectations of comprehensive and transparent public accountability heighten, so too must a hospital's approach to assessing and managing risk. Over a period of two years, the area of patient safety and risk at our hospital has moved from a traditional focus on clinical risk management to an enterprise-wide risk management approach. One of the first community hospitals to embrace enterprise risk management (ERM), St. Joseph's Health Centre, in Toronto, Ontario, has seen early benefits in this transformational journey. This article discusses our approach to the development of an ERM program, tools used and lessons learned.


Neuroendocrine Tumors of the Fallopian Tube: Report of a Case Series and Review of the Literature.
Primary neuroendocrine tumors of the fallopian tube are extremely rare with a few reported cases of high-grade neuroendocrine carcinoma and a single report of a carcinoid tumor arising in a teratoma. We report 4 cases of probable primary neuroendocrine tumors of the fallopian tube (2 carcinoid tumors/low-grade neuroendocrine tumors and 2 high-grade neuroendocrine carcinomas) in patients aged 49 to 71. These represent the first reported cases of primary tubal carcinoid tumor unassociated with a teratoma. We review the published literature regarding primary neuroendocrine tumors of the fallopian tube and speculate on the possible histogenesis of these neoplasms.


The nonlymphoid microenvironment of reactive follicles and lymphomas of follicular origin as defined by immunohistology on paraffin-embedded tissues.
Twenty-five reactive lymph nodes, 10 palatine tonsils, and 72 B-cell non-Hodgkin's lymphomas (NHLs) of supposed follicular origin were investigated in an immunohistologic study of fixed, paraffin-embedded tissues using a panel of monoclonal antibodies reactive with antigens resistant against fixation and paraffin-embedding techniques together with polyclonal antibodies. The results concerning the microenvironmental organization of reactive lymphoid follicles confirmed that the distribution of CD21+ and CD23+ dendritic reticulum cells, vimentin+ fibroblastic reticulum cells, and CD68+ tingible-body macrophages is heterogeneous with reference to their immunostaining patterns and topographic localization within the germinal center and mantle zone. Moreover, a close microenvironmental similarity between the follicular lymphomas of supposed germinal center or mantle zone origin and their normal counterparts was noted. The study of the microenvironment of the B-zone small lymphocytic lymphoma cases, showing the same distribution patterns for the nonlymphoid cells as seen in mantle zone lymphomas, corroborated the supposed follicular origin of this unusual variant of small lymphocytic lymphoma. In conclusion, this study shows that monoclonal antibodies recognizing CD21, CD23, and CD68 antigens may be valuable additions to vimentin, S-100 protein, laminin, and type IV collagen antibodies for investigating the microenvironmental organization of lymphoid tissues in both normal and neoplastic conditions.


Continuous-Flow Synthesis of (R)-Tamsulosin Utilizing Sequential Heterogeneous Catalysis.
We describe the continuous-flow synthesis of (R)-tamsulosin, a blockbuster therapeutic drug employed for dysuria associated with urinary stones and benign prostatic hyperplasia, by utilizing sequential heterogeneous catalysis. Two heterogeneous catalysts have been developed for the synthesis, and the key step involves reductive amination of nitriles using dimethylpolysilane-modified Pd on activated carbon/calcium phosphate. Overall, (R)-tamsulosin was obtained in 60 % yield and 64 % ee (99 % ee after recrystallization) in a flow stream through four catalytic transformations without the need for the isolation or purification of any intermediates or byproduct.


Genetic and Antigenic Diversity of Infectious Bronchitis Virus in South America.
Infectious bronchitis virus (IBV) is one of the economically most important diseases affecting the South American poultry industry. The extensive genomic heterogeneity of IBV is a consequence of high mutation rates and recombination events followed by selection. Nucleotide heterogeneity is much higher in the S1 coding region of the relevant spike protein; thus, the S1 sequence is widely used for the IBV genetic classification in genotypes and lineages. Two main lineages (GI-11 and GI-16) extensively circulate in South American chicken flocks. The GI-11 lineage, found exclusively in South America, emerged in the 1950s and is currently the predominant lineage in Brazil and Uruguay. The GI-16 lineage emerged around 1979 and is now circulating in most South American regions. All South American countries include Massachusetts-type strains (GI-1 lineage) in the IBV vaccination programs. The GI-11 and GI-16 lineages display very low antigenic relatedness to Massachusetts vaccine strains. Because these vaccine strains may not confer complete protection against South American lineages, other vaccination strategies have been reported to control GI-11 and GI-16 outbreaks. Analysis of the few full-length genomes of South American strains highlights a complex recombination history of IBV in the continent. A broader geographic and temporal sampling is needed to understand the pattern of genetic variability and the evolutionary history of IBV variants in South America.


Fully computational lead-development of fluoro-olefin polymerization catalysts.
Coordinative polymerization of fluorinated olefins is still a challenging task. We analyzed the catalytic properties of diimido chromium VI compounds computationally by density functional methods. It was found that reactivity predictions depend strongly on the density functional chosen for the computations. Therefore, DFT calculations were calibrated to high level wave function theory calculations. Then geometrical parameters, which can be tuned by a ligand, were scanned to elucidate their influence on the catalytic activity. Having arrived at an optimal parameter set, we randomly assembled chromium complexes from a fragments database and automatically checked for their activities. This led to the proposal of three exemplary catalyst candidates.


Increased Diameters of the Internal Cerebral Veins and the Basal Veins of Rosenthal Are Associated with White Matter Hyperintensity Volume.
White matter hyperintensities on T2-weighted MR imaging are typical in older adults and have been linked to several poor health outcomes, including cognitive impairment and Alzheimer disease. The presence and severity of white matter hyperintensities have traditionally been attributed to occlusive arteriopathy, but recent evidence also implicates deep medullary venule collagenosis and associated vasogenic edema. Historically, postmortem analyses have been the sole way to analyze cerebral veins, but SWI can be now used to examine cortical veins in vivo. The aim of the current study was to determine whether there is an association between the diameters of the large draining cerebral veins/sinuses and white matter hyperintensity volume. T2-weighted FLAIR and SWI were performed in 682 older adults without dementia (mean age, 73.9 ± 5.9 years; 59.1% women). Total and regional white matter hyperintensity volume was derived. We measured the diameters of 5 regions of the cerebral venous draining system: internal cerebral veins, basal veins of Rosenthal, superior sagittal sinus, vein of Galen, and straight sinus terminus. Increased diameter of the internal cerebral veins was associated with greater total white matter hyperintensity volume (β = 0.09, P = .02) and regionally in the parietal (β = 0.10, P = .006), frontal (β = 0.09, P = .02), and temporal (β = 0.09, P = .02) lobes. Increased diameter of the basal veins of Rosenthal was associated with greater total (β = 0.10, P = .01), frontal (β = 0.11, P = .003), and temporal (β = 0.09, P = .02) white matter hyperintensity volume. Our results suggest that the caliber of the internal cerebral veins and of the basal veins of Rosenthal relates to regional white matter disease.


The Placenta in Congenital Heart Disease: Form, Function and Outcomes.
The maternal-fetal environment, controlled and modulated by the placenta, plays a critical role in the development and well-being of the fetus, with long-term impact through programming of lifelong health. The fetal cardiovascular system and placenta emerge at the same time embryologically, and thus placental form and function are altered in the presence of congenital heart disease (CHD). In this review, we report on what is known about the placenta from a structural and functional perspective when there is CHD. We describe the various unique pathologic findings as well as the diagnostic imaging tools used to characterize placental function in utero. With growing interest in the placenta, a standardized approach to characterizing placental pathology has emerged. Furthermore, application of ultrasonography techniques and magnetic resonance imaging now allow for insights into placental blood flow and functionality in vivo. An improved understanding of the intriguing relationship between the placenta and the fetal cardiovascular system will provide opportunities to develop novel ways to optimize outcomes. Once better understood, therapeutic modulation of placental function offered during the vulnerable period of fetal plasticity may be one of the most impactful ways to alter the course of CHD and its complications.


Catastrophic cervical spine injuries in the collision sport athlete, part 2: principles of emergency care.
Catastrophic cervical spine injuries can lead to devastating consequences for the collision athlete. Improved understanding of these injuries can lead to identification of risk factors, early diagnosis, and effective on-field management. This article is the second in a 2-part series. The first part, published in the June 2004 issue, reviewed the current concepts regarding the epidemiology, functional anatomy, and diagnostic considerations relevant to cervical spine trauma in collision sports. In this article, the principles of on-field emergency care of the spine-injured athlete are reviewed. The authors discuss the need for effective pre-event planning, on-field evaluation and management of cervical spine injuries, and the transition of care from the playing field to the emergency room. The protocol for equipment removal, when necessary, is also reviewed. An organized, rapid approach to the management of cervical spine-injured collision athletes can help to optimize the outcomes of these catastrophic injuries.


Three-dimensional culture of endometrial cells: an in vitro model of endometriosis.
Endometriosis is the presence of endometrial tissue outside of the uterine cavity and is the most common gynecologic disorder in women of reproductive age. Although the quality of life for women with endometriosis is severely compromised, very little is known about the pathophysiology of endometriosis and current therapeutic strategies provide temporary symptomatic relief but not a cure. Endometriosis remains poorly understood primarily because of an inability to identify patients with early stage disease. Animal models have been developed to study early endometriosis but all have some problems that limit their usefulness in determination of the pathophysiology of endometriosis as it occurs in the human. We have preliminary evidence that in the presence of a three-dimensional fibrin matrix, human endometrial glands, stroma, and neovascularization can develop in vitro, mimicking the earliest stages of endometriosis. We believe this model system reflects the situation in the peritoneal cavity of women following retrograde menstruation when endometrial fragments, fibrin, leucocytes and cytokines are trapped in pockets in the dependent parts of the pelvis, allowing endometrial cell proliferation, invasion and angiogenesis to occur. In the present review article, we will further discuss this in vitro model of early endometriosis and discuss possible anti-angiogenic drugs that are already commercially available in an attempt to find an effective and specific treatment for endometriosis.


[Study of the mouse liver microsomes by the methods of proteome analysis].
Proteome maps of microsomes and their ghosts (i.e. membranes purified from "ballast" proteins) were obtained using highly purified mouse liver microsomes. Comparative analysis of protein composition of ghosts without and after the induction with phenobarbital (cytochromes P450 inducer) by using 1D- and 2D-electrophoresis and MALDI-TOF-mass-spectrometry revealed more than 30 new proteins, in the course of induction in the 45-60 kDa range (corresponding to the mol. weights of cytochromes P450). In the 17 kDa range (corresponding to the mol. wt. of cytochrome b5) there were 4 additional protein stains about 20 proteins disappeared over the entire electrophoregram). Separation of microsomal ghosts proteins by 1D electrophoresis followed by mass-spectrometric analysis allowed to identify cytochromes P450. The present investigation demonstrates the efficiency of different proteomic methods combination (1D- and 2D-electrophoresis, mass-spectrometry, bioinformatics and determination of the enzyme activities) for cytochromes P450 identification and elucidation of their functioning in different animal tissues and then extrapolating this approach to humans.


Diabetic amyotrophy progressing to severe quadriparesis.
Diabetic amyotrophy is a distinctive form of diabetic neuropathy usually characterized by the abrupt onset of pain and asymmetric proximal leg weakness and wasting. Involvement of the upper limbs is unusual, and prognosis is said to be good. We describe two patients, each with type II diabetes mellitus, who presented with diabetic amyotrophy progressing to severe quadriparesis. One patient remains severely disabled. The clinical spectrum of diabetic amyotrophy includes progression to severe quadriparesis.


Barometric pressure and triaxial accelerometry-based falls event detection.
Falls and fall related injuries are a significant cause of morbidity, disability, and health care utilization, particularly among the age group of 65 years and over. The ability to detect falls events in an unsupervised manner would lead to improved prognoses for falls victims. Several wearable accelerometry and gyroscope-based falls detection devices have been described in the literature; however, they all suffer from unacceptable false positive rates. This paper investigates the augmentation of such systems with a barometric pressure sensor, as a surrogate measure of altitude, to assist in discriminating real fall events from normal activities of daily living. The acceleration and air pressure data are recorded using a wearable device attached to the subject's waist and analyzed offline. The study incorporates several protocols including simulated falls onto a mattress and simulated activities of daily living, in a cohort of 20 young healthy volunteers (12 male and 8 female; age: 23.7 ±3.0 years). A heuristically trained decision tree classifier is used to label suspected falls. The proposed system demonstrated considerable improvements in comparison to an existing accelerometry-based technique; showing an accuracy, sensitivity and specificity of 96.9%, 97.5%, and 96.5%, respectively, in the indoor environment, with no false positives generated during extended testing during activities of daily living. This is compared to 85.3%, 75%, and 91.5% for the same measures, respectively, when using accelerometry alone. The increased specificity of this system may enhance the usage of falls detectors among the elderly population.


Investigating the role of ischemia vs. elevated hydrostatic pressure associated with acute obstructive uropathy.
Obstructive uropathy can cause irreversible renal damage. It has been hypothesized that elevated hydrostatic pressure within renal tubules and/or renal ischemia contributes to cellular injury following obstruction. However, these assaults are essentially impossible to isolate in vivo. Therefore, we developed a novel pressure system to evaluate the isolated and coordinated effects of elevated hydrostatic pressure and ischemic insults on renal cells in vitro. Cells were subjected to: (1) elevated hydrostatic pressure (80 cm H(2)O); (2) ischemic insults (hypoxia (0% O(2)), hypercapnia (20% CO(2)), and 0 mM glucose media); and (3) elevated pressure + ischemic insults. Cellular responses including cell density, lactate dehydrogenase (LDH) release, and intracellular LDH (LDH(i)), were recorded after 24 h of insult and following recovery. Data were analyzed to assess the primary effects of ischemic insults and elevated pressure. Unlike pressure, ischemic insults exerted a primary effect on nearly all response measurements. We also evaluated the data for insult interactions and identified significant interactions between ischemic insults and pressure. Altogether, findings indicate that pressure may sub-lethally effect cells and alter cellular metabolism (LDH(i)) and membrane properties. Results suggest that renal ischemia may be the primary, but not the sole, cause of cellular injury induced by obstructive uropathy.


[Pulsatile release of gonadotropins in athletic women].
In athletic women menstrual disorders such as luteal insufficiency, oligomenorrhea and amenorrhea are often seen. It has been suggested that such disorders may be related to hypothalamic-pituitary axis insufficiency caused by physical activity. To investigate the mechanism by which the disorder is promoted, episodic gonadotropin secretion was studied in 10 athletic women (normal ovulatory, 5; short luteal, 5) and 6 non-athletic controls. In the early follicular phase of the menstrual cycle, blood samples were obtained through an indwelling venous catheter at 15-minute intervals for 4 hours. The concentrations of luteinizing hormone (LH) and follicle-stimulating hormones (FSH) were measured by radioimmunoassay. The mean LH and FSH concentrations in the athletic groups were lower than those in the control group. LH pulse frequencies in the short luteal group were 2.8 +/- 0.2 times/four hours (mean +/- standard error) and decreased compared with that of the control group (3.7 +/- 0.2 times/four hours; p less than 0.001). The LH pulse amplitudes of all three groups were similar. Pulsatile FSH secretion was also observed in all the women. Dynamism of FSH was the same as that of LH except for low pulse amplitude in the athletic groups. These data suggest that hypothalamic-pituitary axis insufficiency, especially of hypothalamic origin may be caused by athletic activity.


Philometra (Ranjhinema) salgadoi n. sp. (Nematoda: Philometridae) from the ocular cavity of the red grouper Epinephelus morio (Pisces: Serranidae) from the yucatan peninsula, Mexico.
A new nematode species, Philometra (Ranjhinema) salgadoi n. sp., is described from females found in the ocular cavity of the red grouper (Epinephelus morio) from the Mexican coast of the Gulf of Mexico (type locality Telchac, Yucatán). The nematode is characterized by the presence of 8 conspicuously large, crescent-shaped, fleshy cephalic papillae of the external circle and 2 small, subterminal papilla-like projections on the caudal end. Another philometrid species, Philometroides belousae n. sp., is established by indication for P. parasiluri (Yamaguti, 1935) sensu Ermolenko, 1984; its adult females are parasitic in the subcutaneous tissues of Percottus glehni in the Russian Far East.


A novel cytoplasmic hemimethylated oriC binding activity.
Using hemimethylated, fully methylated, and unmethylated oligonucleotide probes corresponding to part of the origin of Escherichia coli DNA replication, oriC (+81-136), we have characterized a novel hemimethylated DNA-specific protein binding activity. This activity appears to be located in the cytoplasm rather than in membrane fractions. It has been partially purified and, in DNase footprinting analysis, found to preferentially protect only a subset of the hemimethylated GATC sites present in the minimal oriC. These sites are found adjacent to the DnaA binding box, R1, and overlap the integration host factor binding site. The activity does not correspond to known hemimethylated binding proteins, although in the seqA deletion mutant, there is a 3-fold reduction of the activity. The stage of the cell cycle in synchronized PC2 cultures does not seem to significantly affect thte relative levels of this binding activity. A possible role in sequestration of the newly replicated hemimethylated origin is discussed.


A new Paracoccidioides brasiliensis 70-kDa heat shock protein reacts with sera from paracoccidioidomycosis patients.
A cDNA coding for a new member of the 70-kDa heat shock proteins (HSP70) family from the dimorphic and pathogenic fungus, Paracoccidioides brasiliensis, was cloned and characterized. The cDNA-deduced sequence coded for 655 amino acid residues and showed 95% identity to a previously described P. brasiliensis hsp70 gene. Cytoplasmic and typical nuclear localization signals, which indicate induction upon stress, were identified in the deduced peptide. The complete hsp70 cDNA coding region was cloned into a pGEX 4T-3 plasmid and expressed in Escherichia coli as a glutathione-S-transferase-tagged fusion protein. The recombinant protein reacted with a rabbit polyclonal antibody against HSP70. Western immunoblot experiments demonstrated that sera from paracoccidioidomycosis patients recognized the purified recombinant protein, suggesting an immunological role for this protein in the infectious process. The antigenicity analysis of rHSP70 detected three internal peptides that could act as activators of T-cell proliferation.


The Prevalence of Polypharmacy and Potentially Inappropriate Medications and Its Relationship with Cognitive Status in Portuguese Institutionalized Older Adults: A Cross-Sectional Study.
The aim of this study was to evaluate the prevalence of polypharmacy and potentially inappropriate medications (PIMs) in a population of older adults living in nursing homes. Furthermore, we also intended to assess the possible association between polypharmacy, potentially inappropriate medications and cognitive impairment in institutionalized older adults. A cross-sectional study analyzed data from 193 nursing home residents in the district of Viseu, Portugal, between September 2018 and June 2019, with a mean age of 82.4 ± 6.2 years (ranging from 65 to 95 years old); 72.5% (n = 140) were female participants. Major polypharmacy was presented in 80.8% of the study population, who took 7.6 ± 3.3 drugs per day. Using the Beers Criteria, we found that 79.3% took PIMs. There was a positive association between polypharmacy and PIM (p < 0.001), showing that higher medicines intake increased the number of PIMs. Polypharmacy was not associated with the functionality of the older adults to perform activities of daily living, but was associated with cognitive impairment. The older adults with lower scores on the Mini Mental State Examination (MMSE) took more drugs (p = 0.039) and used more PIM (p < 0.001). Moreover, patients taking five or more prescription drugs per day (major polypharmacy) consuming any psychiatric, gastrointestinal or oral antidiabetic agents (regardless of whether they were considered potentially inappropriate or not) had higher odds of displaying cognitive impairment than those who did not (p < 0.05). Older adult residents of the studied nursing homes were potentially affected by polypharmacy and inappropriate polypharmacy. This observation reveals the need to adopt and implement strategies that make drug therapy more adequate and safer for older adults.


The potential of ecosystem-based management to integrate biodiversity conservation and ecosystem service provision in aquatic ecosystems.
Global aquatic biodiversity keeps declining rapidly, despite international efforts providing a variety of policies and legislations that identify goals for, and give directions to protecting the world's aquatic fauna and flora. With the H2020 project AQUACROSS, we have made an unprecedented effort to unify policy strategies, knowledge, and management concepts of freshwater, coastal, and marine ecosystems to support the achievement of the targets set by the EU Biodiversity Strategy to 2020. AQUACROSS has embraced the concept of ecosystem-based management (EBM), which approaches environmental management from a social-ecological system perspective to protect biodiversity and to sustainably harvest ecosystem services. This special issue includes contributions resulting from AQUACROSS, which either tackle selected EBM challenges from a theoretical point of view or apply EBM in one of the selected case studies across Europe. In this article, we introduce relevant topics, address the most important lessons learnt, and suggest where research should go with aquatic EBM. We hope that this special issue will foster and facilitate the uptake of EBM in aquatic ecosystems and, therewith, provide the on-ground applications needed for evaluating EBM's utility to safeguard aquatic biodiversity.


Thrombolysis in acute ischemic stroke: successes, failures, and new hopes.
Evidence from randomized clinical trials indicates that systemic administration of recombinant tissue plasminogen activator (rtPA) is a highly effective treatment for acute ischemic stroke, provided that treatment is administered within the first 3 h after stroke onset. An absolute increase in favorable outcome of up to 13% has been reported, and a pooled analysis of six randomized trials has shown that, although the sooner rtPA is given the greater the benefit, efficacy is present up to 4.5 h after stroke onset. Despite of the spreading use of tPA in different countries and continents, there are still a number of burdens and failures in the optimal accomplishment of thrombolytic treatment. rtPA is used in less than 4% of patients, reperfusion and complete recovery is achieved in less than 50% of patients, and treatment is denied to many patients. However, important advances in clinical investigation suggest that new aims and hopes will be achieved in the near future. Ultrasound-enhanced systemic thrombolysis, the use of MRI for selecting acute stroke patients for IV or IA thrombolysis after 3 h, mechanical embolus disruption or removal in proximal artery occlusions, and the potential usefulness of new biomarkers of blood brain barrier disruption and hemorrhagic risk are promising strategies that may improve the risk/benefit ratio and increase the number of patients who will benefit from thrombolytic therapy.


Angiotensin II action in isolated microperfused rabbit afferent arterioles is modulated by flow.
We have recently presented evidence that endogenous nitric oxide (NO) and prostaglandins (PGs) modulate angiotensin II (Ang II) action in microperfused afferent arterioles (Af-Arts). Because flow may be a physiological stimulus of endothelial release of NO and PGs, we tested the hypothesis that flow through the lumen of the Af-Art stimulates the endothelium to produce NO and PGs, which in turn modulate the action of Ang II. We microdissected the terminal segment of an interlobular artery together with two Af-Arts, their glomeruli and efferent arterioles (Ef-Art). The two Af-Arts were perfused simultaneously from the interlobular artery, while one Ef-Art was occluded. Since the arteriolar perfusate contained 5% albumin, oncotic pressure built up in the glomerulus with the occluded Ef-Art and opposed the force of filtration, resulting in little or no flow through the corresponding Af-Art. Thus this preparation allowed us to observe Ang II action in free-flow and non-flow Af-Arts simultaneously. Ang II-induced constriction was weaker in free-flow than non-flow Af-Arts, with the luminal diameter decreasing by 8 +/- 2% and 23 +/- 3% at 10(-9) M, respectively (P < 0.013 free-flow vs. non-flow; N = 9). Disrupting the endothelium augmented Ang II action in free-flow (33 +/- 5.1%; P < 0.01 vs. intact endothelium) but not non-flow Af-Arts (31 +/- 5.3%), thus abolishing the differences between them (N = 8). Pretreatment with an inhibitor of either NO synthase (N-nitro-L-arginine methyl ester) or cyclooxygenase (indomethacin) augmented Ang II action more in free-flow than non-flow Af-Arts, likewise abolishing the differences between them. These results suggest that intraluminal flow modulates the vasoconstrictor action of Ang II in Af-Arts via endothelium-derived NO and PGs. Thus flow may be important in the fine control of glomerular hemodynamics.