Chemical and thermal unfolding of calreticulin.
Calreticulin is a soluble endoplasmic reticulum chaperone, which has a relatively low melting point due to its remarkable structure with a relatively high content of flexible structural elements. Using far ultraviolet circular dichroism (CD) spectroscopy and a fluorescent dye binding thermal shift assay, we have investigated the chemical and thermal stability of calreticulin. When the chemical stability of calreticulin was assessed, a midpoint for calreticulin unfolding was calculated to 3.0M urea using CD data at 222 nm. Using the fluorescent dye binding thermal shift assay, calreticulin was found to obtain a molten structure in urea concentrations between 1-1.5 M urea, and to unfold/aggregate at high and low pH values. The results demonstrated that the fluorescent dye binding assay could measure the thermal stability of calreticulin in aqueous buffers with results comparable to melting points obtained by other techniques.


Antiviral characteristics of GSK1265744, an HIV integrase inhibitor dosed orally or by long-acting injection.
GSK1265744 is a new HIV integrase strand transfer inhibitor (INSTI) engineered to deliver efficient antiviral activity with a once-daily, low-milligram dose that does not require a pharmacokinetic booster. The in vitro antiviral profile and mechanism of action of GSK1265744 were established through integrase enzyme assays, resistance passage experiments, and cellular assays with site-directed molecular (SDM) HIV clones resistant to other classes of anti-HIV-1 agents and earlier INSTIs. GSK1265744 inhibited HIV replication with low or subnanomolar efficacy and with a selectivity index of at least 22,000 under the same culture conditions. The protein-adjusted half-maximal inhibitory concentration (PA-EC50) extrapolated to 100% human serum was 102 nM. When the virus was passaged in the presence of GSK1265744, highly resistant mutants with more than a 10-fold change (FC) in EC50 relative to that of the wild-type were not observed for up to 112 days of culture. GSK1265744 demonstrated activity against SDM clones containing the raltegravir (RAL)-resistant Y143R, Q148K, N155H, and G140S/Q148H signature variants (FC less than 6.1), while these mutants had a high FC in the EC50 for RAL (11 to >130). Either additive or synergistic effects were observed when GSK1265744 was tested in combination with representative anti-HIV agents, and no antagonistic effects were seen. These findings demonstrate that, similar to dolutegravir, GSK1265744 is differentiated as a new INSTI, having a markedly distinct resistance profile compared with earlier INSTIs, RAL, and elvitegravir (EVG). The collective data set supports further clinical development of GSK1265744.


Increased granulocyte colony-stimulating factor responsiveness but normal resting granulopoiesis in mice carrying a targeted granulocyte colony-stimulating factor receptor mutation derived from a patient with severe congenital neutropenia.
The role of mutations of the granulocyte colony-stimulating factor receptor (G-CSFR) in the pathogenesis of severe congenital neutropenia (SCN) and the subsequent development of acute myeloid leukemia (AML) is controversial. Mice carrying a targeted mutation of their G-CSFR that reproduces the mutation found in a patient with SCN and AML have been generated. The mutant G-CSFR allele is expressed in a myeloid-specific fashion at levels comparable to the wild-type allele. Mice heterozygous or homozygous for this mutation have normal levels of circulating neutrophils and no evidence for a block in myeloid maturation, indicating that resting granulopoiesis is normal. However, in response to G-CSF treatment, these mice demonstrate a significantly greater fold increase in the level of circulating neutrophils. This effect appears to be due to increased neutrophil production as the absolute number of G-CSF-responsive progenitors in the bone marrow and their proliferation in response to G-CSF is increased. Furthermore, the in vitro survival and G-CSF-dependent suppression of apoptosis of mutant neutrophils are normal. Despite this evidence for a hyperproliferative response to G-CSF, no cases of AML have been detected to date. These data demonstrate that the G-CSFR mutation found in patients with SCN is not sufficient to induce an SCN phenotype or AML in mice.


2-Triazenoazaindoles: α novel class of triazenes inducing transcriptional down-regulation of EGFR and HER-2 in human pancreatic cancer cells.
Pancreatic cancer is a complex malignancy arising from the accumulation of genetic and epigenetic defects in the affected cells. Standard chemotherapy for patients with advanced disease shows only modest effects and is associated with considerable toxicity. Overexpression or aberrant activation of members of the epidermal growth factor receptor tyrosine kinase family, which includes EGFR and HER-2, occurs frequently and is associated with multiple drug resistance and decreased patient survival. In this study, we have investigated the therapeutic potential of AS104, a novel compound of the triazene class, with potential inhibitory effects on EGFR. We found that treatment of cells with AS104 causes significant reduction of cell growth and metabolic activity in four human pancreatic cancer cell lines. Furthermore, we show that the AS104-mediated induction of apoptotic cell death is associated with stimulation of autophagy in a dose-dependent manner. Treatment of cells with AS104 results in significant down-regulation of EGFR and HER-2 expression and activity and subsequent inhibition of downstream signaling proteins. Quantitative RT-PCR analysis and assays with proteasome inhibitors revealed that AS104 regulates the expression of EGFR and HER-2 at the transcriptional level. These findings provide for the first time experimental evidence for efficacy of AS104 in the simultaneous transcriptional repression of EGFR and HER-2 genes and suggest that AS104 may have therapeutic potential in the treatment of pancreatic cancers that express high levels of the aforementioned receptor tyrosine kinases.


Characterization of the protease processing sites in a multidomain proteinase inhibitor precursor from Nicotiana alata.
A gene encoding a 40.3-kDa serine proteinase inhibitor (PI) precursor is expressed at high levels in the stigma of the ornamental tobacco, Nicotiana alata. The precursor is processed proteolytically in vivo to release five homologous proteinase inhibitors of approximately 6 kDa, as well as two flanking peptides. The five PIs have been purified from stigmas and identified by N-terminal sequencing, electrospray mass spectrometry and inhibition activity against chymotrypsin or trypsin. One of the PIs inhibits chymotrypsin and the other four are most active on trypsin. Cleavage occurs in a linker region (EEKKND) that is repeated six times in the precursor molecule. In the plant, the initial cleavage probably occurs between asparagine and the aspartate residues and ragged ends are formed by subsequent trimming. In vitro, the protease-sensitive linker region is selectively cleaved by the endoproteinases Asp-N, Glu-C and Lys-C to release fully active approximately 6-kDa PIs that are resistant to further proteolytic digestion. The precursor, produced by a recombinant baculovirus, inhibits chymotrypsin more effectively than trypsin. The stoichiometry of 2.6 trypsin molecules/1 precursor molecule indicates that processing is required to activate or expose all of the four trypsin inhibitory sites.


Critical Reviews: How we treat sickle cell patients with leg ulcers.
The past five decades have seen an improvement in the mortality and morbidity of sickle cell disease (SCD) because of prophylaxis against infectious complications, improved and expanded red cell transfusions, implementation of hydroxyurea therapy, and advances in supportive care. Now that the majority of patients in the western hemisphere reaches adulthood, end organ diseases are frequent, which include vasculopathic complications such as chronic leg ulcers. The management of patients with leg ulcers requires the hematologist to lead a team of health care professionals, and investigates the presence of associated, but potentially still occult signs of vasculopathy, such as pulmonary hypertension, renal disease, priapism and retinopathy. These complications may be asynchronous, and long term careful screening is indicated, in order to ensure early diagnosis and intervention. It is crucial to address both the immediate consequences of pain, infection and disability, and long term effects on quality of life, employment and stigma associated with chronic ulceration. Recent insights into their pathophysiology may have practical implications. We propose a holistic approach to the management of patients' physical and emotional problems and mechanisms of ulcers formation and delayed healing. An overview of topical and systemic therapies for chronic ulcers is given, with the understanding that wound care therapy is best left to the wound specialists, medical and surgical, with whom the hematologist must keep an open line of communication. In the absence of evidence-based guidelines, our opinion is based on both a critical review of the literature and our personal clinical and research experience.


MFV-class: a multi-faceted visualization tool of object classes.
Classes are key software components in an object-oriented software system. In many industrial OO software systems, there are some classes that have complicated structure and relationships. So in the processes of software maintenance, testing, software reengineering, software reuse and software restructure, it is a challenge for software engineers to understand these classes thoroughly. This paper proposes a class comprehension model based on constructivist learning theory, and implements a software visualization tool (MFV-Class) to help in the comprehension of a class. The tool provides multiple views of class to uncover manifold facets of class contents. It enables visualizing three object-oriented metrics of classes to help users focus on the understanding process. A case study was conducted to evaluate our approach and the toolkit.


A novel homozygous variant in REN in a family presenting with classic features of disorders involving the renin-angiotensin pathway, without renal tubular dysgenesis.
Autosomal recessively inherited pathogenic variants in genes associated with the renin-angiotensin-aldosterone system (RAAS) result in early onset oligohydramnios and clinical features of the Potter sequence, typically in association with proximal renal tubules dysgenesis. We describe two siblings and a first cousin who had severe oligohydramnios in the second trimester, and presented at birth with loose skin, wide fontanelles and sutures, and pulmonary insufficiency. Two had refractory hypotension during their brief lives and one received palliative care after birth. All were found to have a homozygous nonsense variant, REN: c.891delG; p.Tyr287*, on exome sequencing. Autopsy limited to the genitourinary system in two of the children revealed normal renal tubular histology in both. Immunoblotting confirmed diminished expression of renin within cultured skin fibroblasts. To our knowledge, this is the first identification of an association between biallelic variants in REN and oligohydramnios in the absence of renal tubular dysgenesis. Due to its role in the RAAS, it has previously been proposed that the decreased expression of REN results in hypotension, ischemia, and decreased urine production. We suggest sequencing of genes in the RAAS, including REN, should be considered in cases of severe early onset oligohydramnios, even when renal morphology and histology are normal.


Ribosome Levels Selectively Regulate Translation and Lineage Commitment in Human Hematopoiesis.
Blood cell formation is classically thought to occur through a hierarchical differentiation process, although recent studies have shown that lineage commitment may occur earlier in hematopoietic stem and progenitor cells (HSPCs). The relevance to human blood diseases and the underlying regulation of these refined models remain poorly understood. By studying a genetic blood disorder, Diamond-Blackfan anemia (DBA), where the majority of mutations affect ribosomal proteins and the erythroid lineage is selectively perturbed, we are able to gain mechanistic insight into how lineage commitment is programmed normally and disrupted in disease. We show that in DBA, the pool of available ribosomes is limited, while ribosome composition remains constant. Surprisingly, this global reduction in ribosome levels more profoundly alters translation of a select subset of transcripts. We show how the reduced translation of select transcripts in HSPCs can impair erythroid lineage commitment, illuminating a regulatory role for ribosome levels in cellular differentiation.


Inactivation of stressed Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes in hummus using low dose gamma irradiation.
Hummus is a popular dip in the Middle East region prepared by mixing the boiled chickpeas with tahini and other ingredients, and because its consumption has increased world-wide some notoriety has developed following an increase in the incidence of hummus-related illness outbreaks and recalls. The objectives of the current research were (i) to study the efficiency of low dose gamma irradiation to inhibit Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes in hummus, and (ii) to assess the effect of environmental stresses namely cold, heat, and desiccation on the resistance of these pathogens to gamma irradiation. The samples of hummus were prepared and then individually inoculated with approximately 7.0 log CFU/g of unstressed or cold-, heat-, or desiccated-stressed cocktail cultures of each of E. coli O157:H7, S. enterica, and L. monocytogenes. The inoculated samples were then exposed to gamma irradiation at doses of 0.1 to 0.6 kGy. The numbers of unstressed E. coli O157:H7, S. enterica, and L. monocytogenes were decreased by 0.6-3.9, 0.7-2.9, and 1.0-3.0 log CFU/g, respectively, by irradiation treatment. The resistance of E. coli O157:H7 to gamma irradiation was not affected by desiccation, heat, and cold stresses. However, the pre-exposure of S. enterica and L. monocytogenes cells to these stresses reduced their resistance toward gamma irradiation. PRACTICAL APPLICATION: Gamma irradiation is a non-thermal treatment that can be used in food processing to ensure food safety and quality. The current study proved that low levels (≤0.6 kGy) of gamma irradiation can effectively decrease the risk of unstressed and cold-, heat-, or desiccation-stressed Salmonella enterica, Listeria monocytogenes, or Escherichia coli O157:H7 in hummus.


Acute myocardial infarction complicating ischemic stroke: is there room for cangrelor?
Acute myocardial infarction (AMI) complicating ischemic stroke is a well known and undertreated event. A conservative management is not infrequent in these settings, due to the fear of hemorrhagic complications related to antithrombotic therapy. Notably, an invasive approach with a primary percutaneous coronary intervention (PCI) has been shown to be associated with a lower in-hospital mortality in patients with concomitant ischemic stroke and AMI. The optimal antiplatelet regimen in these cases has been not clearly defined, yet. We report two cases of patients with AMI complicating ischemic stroke, successfully treated with cangrelor infusion, which was started during PCI and maintained up to 48 h at bridge therapy dosage (0.75 mcg/kg/min). Both patients underwent successful PCI in the acute phase, and neither ischemic nor hemorrhagic complications occurred during in-hospital stay.


Influence of age of onset and patient's sex on the prevalence and severity of manifestations of Behçet's syndrome.
Eye disease, arthritis, folliculitis, and thrombophlebitis were more common among males, and erythema nodosum among females, in 297 patients with Behćet syndrome (BS) at their first visit despite an identical disease duration. Younger males and females (age of onset 24 years or less) had a higher prevalence of eye disease and total clinical activity than did the older patients (age of onset 25 years or more). Among the 51 patients followed up for 52 months (SD 7 months) the total clinical activity became significantly less (p less than 0.05) in the whole group at the end of this period. This was particularly true for older females. While male sex and a younger age of onset are associated with more severe disease in BS, overall, the syndrome ran an improving or stable course over 4 1/2 years.


Loss of parietal cell superoxide dismutase leads to gastric oxidative stress and increased injury susceptibility in mice.
Mitochondrial superoxide dismutase (SOD2) prevents accumulation of the superoxide that arises as a consequence of oxidative phosphorylation. However, SOD2 is a target of oxidative/nitrosative inactivation, and reduced SOD2 activity has been demonstrated to contribute to portal hypertensive gastropathy. We investigated the consequences of gastric parietal cell-specific SOD2 deficiency on mitochondrial function and gastric injury susceptibility. Mice expressing Cre recombinase under control of the parietal cell Atpase4b gene promoter were crossed with mice harboring loxP sequences flanking the sod2 gene (SOD2 floxed mice). Cre-positive mice and Cre-negative littermates (controls) were used in studies of SOD2 expression, parietal cell function (ATP synthesis, acid secretion, and mitochondrial enzymatic activity), increased oxidative/nitrosative stress, and gastric susceptibility to acute injury. Parietal cell SOD2 deficiency was accompanied by a 20% (P < 0.05) reduction in total gastric SOD activity and a 93% (P < 0.001) reduction in gastric SOD2 activity. In SOD2-deficient mice, mitochondrial aconitase and ATP synthase activities were impaired by 36% (P < 0.0001) and 44% (P < 0.005), respectively. Gastric tissue ATP content was reduced by 34% (P < 0.002). Basal acid secretion and peak secretagogue (histamine)-induced acid secretion were reduced by 43% (P < 0.0001) and 40% (P < 0.0005), respectively. There was a fourfold (P < 0.02) increase in gastric mucosal apoptosis and 41% (P < 0.001) greater alcohol-induced gastric damage in the parietal cell SOD2-deficient mice. Our findings indicate that loss of parietal cell SOD2 leads to mitochondrial dysfunction, resulting in perturbed energy metabolism, impaired parietal cell function, and increased gastric mucosal oxidative stress. These alterations render the gastric mucosa significantly more susceptible to acute injury.


Discrimination between Gyrodactylus salaris, G. derjavini and G. truttae (Platyhelminthes: Monogenea) using restriction fragment length polymorphisms and an oligonucleotide probe within the small subunit ribosomal RNA gene.
The small subunit ribosomal RNA (srRNA) gene was amplified from Gyrodactylus salaris using the polymerase chain reaction (PCR), cloned, and the complete gene sequence of 1966 bp determined. The V4 region of the srRNA gene was identified and amplified from single specimens of G. salaris, G. derjavini and G. truttae. Comparison of the V4 sequences from these three species revealed sequence differences from which restriction fragment length polymorphisms (RFLPs) were predicted and an oligonucleotide probe (GsV4) specific to G. salaris designed. Digestion of the amplified V4 region of the srRNA gene with Hae III and either Alw I, BstY I, Dde I or Mbo I provided a means of discriminating between G. salaris, G. derjavini and G. truttae. The GsV4 probe was used to detect the srRNA gene from G. salaris in Southern and dot blots of the amplified V4 region.


Point-of-care platelet function testing results in a dog with Bernard-Soulier syndrome.
Bernard-Soulier syndrome (BSS), also known as hemorrhagiparous thrombocytic dystrophy (OMIA 002207-9615), is a rare defect in platelet function recognized in both dogs and humans. It is caused by a deficiency in glycoprotein 1b-IX-V, the platelet surface protein which acts as a receptor for the von Willebrand factor. The characteristic features of BSS in humans and dogs include macrothrombocytes and mild-to-moderate thrombocytopenia with a bleeding tendency. This condition has previously been reported in European Cocker Spaniel dogs; however, the results of platelet function tests in these animals have not been reported. This case report describes a European Cocker Spaniel dog with spontaneously occurring Bernard-Soulier syndrome and the results of point-of-care platelet function tests, including a prolonged buccal mucosal bleeding time (>8 min), prolongation (>300 s) of PFA-200 COL/ADP, COL/EPI, and P2Y closure times, and reduced aggregation (15%-48%) with Plateletworks ADP, but with normal aggregation (92%) with Plateletworks AA. This is the first description of the results of platelet function tests in canine Bernard-Soulier syndrome.


Establishment and characterization of a rat pancreatic stellate cell line by spontaneous immortalization.
Activated pancreatic stellate cells (PSCs) have been implicated in the pathogenesis of pancreatic fibrosis and inflammation. Primary PSCs can be subcultured only several times because of their limited growth potential. A continuous cell line may therefore be valuable in studying molecular mechanisms of these pancreatic disorders. The aim of this study was to establish a cell line of rat PSCs by spontaneous immortalization. PSCs were isolated from the pancreas of male Wistar rats, and conventional subcultivation was performed repeatedly. Telomerase activity was measured using the telomere repeat amplification protocol. Activation of transcription factors was assessed by electrophoretic mobility shift assay. Activation of mitogen-activated protein (MAP) kinases was examined by Western blotting using anti-phosphospecific antibodies. Expression of cytokine-induced neutrophil chemoattractant-1 was determined by enzyme immunoassay. Conventional subcultivation yielded actively growing cells. One clone was obtained after limiting dilution, and designated as SIPS. This cell line has been passaged repeatedly more than 2 years, and is thus likely immortalized. SIPS cells retained morphological characteristics of primary, culture-activated PSCs. SIPS expressed alpha-smooth muscle actin, glial acidic fibrillary protein, vimentin, desmin, type I collagen, fibronectin, and prolyl hydroxylases. Telomerase activity and p53 expression were negative. Proliferation of SIPS cells was serum-dependent, and stimulated with platelet-derived growth factor-BB through the activation of extracellular signal-regulated kinase. Interleukin-1beta activated nuclear factor-kappaB, activator protein-1, and MAP kinases. Interleukin-1beta induced cytokine-induced neutrophil chemoattractant-1 expression through the activation of nuclear factor-kappaB and MAP kinases. SIPS cells can be useful for in vitro studies of cell biology and signal transduction of PSCs.


An epidemiologic study of sacroiliac fusion in some human skeletal remains.
A case-control study was undertaken to generate some hypotheses concerning the etiology of sacroiliac fusion in a group of skeletons dating from the 18th and 19th centuries. Forty-one skeletons with fusion of the sacroiliac joint were compared with eighty-two adult skeletons without the condition. The sacroiliac joints were most frequently fused with bridging osteophytes and no preference for site or side of fusion could be detected. Except where there were other features in the skeleton suggestive of a sero-negative arthropathy, radiography demonstrated that there was no intra-articular ankylosis. We were able to confirm earlier observations that the condition is more prevalent in males and in older age-groups. The study also showed a significant association between sacroiliac fusion and the presence of diffuse idiopathic skeletal hyperostosis (DISH) and osteoarthritis of the spine but not for osteoarthritis at any other site. There was a highly significant association between sacroiliac fusion and the phenomenon that we refer to as "bone forming." We devised a series of bone-former scores and were able to show a significant excess of cases with high scores compared with the controls. This association persisted when allowance had been made for potential confounding factors such as DISH, osteoarthritis of the spine, and age.


The Hidden Burden of Plasmodium vivax Malaria in Pregnancy in the Amazon: An Observational Study in Northwestern Brazil.
We measured the prevalence of malaria in pregnancy and estimated its impact on birth weight and length and maternal hemoglobin in 1,180 women from Juruá Valley, the main malaria hotspot in Brazil. Antenatal malaria episodes, 74.6% of them due to Plasmodium vivax, were microscopically diagnosed in 8.0% of the women and were associated with an average reduction in birth weight z-scores of 0.35 (95% confidence interval [CI] = 0.14-0.57) and in birth length z-scores of 0.31 (95% CI = 0.08-0.54), compared with malaria-free pregnancies. Affected mothers had a mean decrease in hemoglobin concentration at delivery of 0.33 g/100 mL (95% CI = 0.05-0.62 g/100 mL); 51.6% were anemic. The timing and frequency of antenatal infections influenced pregnancy outcomes and first- or second-trimester infections were not associated with decreased birth weight and length and maternal hemoglobin at delivery. Although repeated antenatal vivax infections were associated with poorer birth outcomes, even a single vivax malaria episode was associated with a significant reduction in birth weight and length and maternal hemoglobin. Overall, 7.5% women had the parasite's DNA found in peripheral blood at delivery. Most (83.1%) of these 89 perinatal infections were due to P. vivax and only 7.9% of them progressed to symptomatic disease after delivery. Plasmodium vivax and Plasmodium falciparum DNA was found in 0.6% and 0.3% of 637 cord blood samples examined, respectively, but only one newborn developed clinical neonatal malaria. Our results further challenge the notion that vivax malaria is relatively benign during pregnancy and call for better strategies for its prevention.


Interference and facilitation in overt speech production investigated with event-related potentials.
We report an event-related potential study investigating the neural basis of interference and facilitation in the picture-word interference paradigm with immediate overt naming. We used the high temporal resolution of the electrophysiological response to dissociate general and specific interference processes, by comparing unrelated word distractors to nonlinguistic (a row of Xs), surface feature denoting, and category member distractors. Our results first indicate that the increased naming latencies for linguistic relative to nonlinguistic distractors are because of general conflict-monitoring processes, associated with early event-related potential effects (120-220 ms) and increased activity in the anterior cingulate cortex. Next, distractors specifying a surface feature of the picture seem to facilitate its identification within the same time window, which involves widespread networks. Finally, nonlinguistic and surface feature distractors also reduced the N400 amplitude, relative to unrelated word distractors. Taken together our results support the view that several distinct processes give rise to the reaction time results often observed in picture naming.


Expression of proliferation markers Ki67, cyclin A, geminin and aurora-kinase A in primary breast carcinomas and corresponding distant metastases.
To assess the expression of the following cell cycle regulatory proteins in primary metastatic breast carcinomas (MBCs) and on availability in matched distant metastases (DMs): Ki67, cyclin A, geminin and aurora-kinase A (aurkA); and to compare the expression of these markers in early MBC (EMBC) and late MBC separated into groups according to median time point on metastatic event occurred (28 months). The expression of the above mentioned markers was analysed in a total of 47 primary MBCs and 59 DMs (out of which 37 were pairs) by immunohistochemistry. Fourteen breast carcinomas with no relapse over a 10-year follow-up period were utilised as control cases (CBC). Among the MBCs, 22 metastasised to the bone, 4 to the lung and 21 to the central nervous system (CNS). Geminin (p<0.001) and Ki67 (p=0.001) were increased in the MBCs while aurkA and cyclin A showed no difference when compared with CBCs. There were no differences between aurkA, cyclin A and geminin expression in MBCs and DMs in general. Expression of Ki67 was, however, elevated (p=0.027) in DMs. In CNS metastases all markers showed elevated expression as compared to MBCs. In bone metastases, geminin was lower (p<0.001) compared with primary MBCs. In the metastases of the lung, the evaluated markers did not show different expression. According to the median follow-up until the metastatic event, Ki67 was found to be significantly elevated in EMBC (p=0.018). Ki67 index and geminin distinguish a fraction of MBC with worse prognosis, showing increased levels in the latter in comparison to CBC being tumour-free over a 10-year follow-up period. Ki67 could possibly identify a group of MBCs that develop early DMs.


Nucleotide sequence of the hepatitis B virus genome (subtype ayw) cloned in E. coli.
The complete nucleotide sequence of hepatitis B virus genome (subtype ayw) cloned in Escherichia coli has been determined using the Maxam and Gilbert method and the dideoxynucleotide method. This sequence is 3,182 nucleotides long. Location of the nonsense codons shows that the coding capacity of the L chain is larger than the coding capacity of the S chain. Eight open regions, able to code for polypeptide chains larger than 100 amino acids, have been located. Region 6, which is the largest, covers more than 80% of the genome. The gene S which codes for polypeptide I of the Hbs Ag and was previously located between coordinates 95.1 and 73.6 is contained in region 7.


Novel combinatorial selection of phosphorothioate oligonucleotide aptamers.
A novel combinatorial approach is described for construction and screening of enhanced nuclease-resistant phosphorothioate DNA "decoys" or "aptamers." Aptamers have been selected that bind tightly to the nuclear factor for human IL6 (NF-IL6), a basic leucine zipper transcription factor involved in the induction of acute-phase responsive and cytokine gene promotors in response to inflammation. Using a random combinatorial selection approach and dNTP(alphaS)'s in the PCR amplification, we can select specific thio-substituted agents which have the highest specificity in binding to target NF-IL6. Using a 22-nucleotide-long duplex random library, nanomolar binding, specific 22-mer thiophosphate backbone substitution sequences (at dA positions only) were selected. These show a different consensus sequence than normal phosphate backbone CCAAT/enhancer binding protein recognition sites for NF-IL6. Unlike the wild-type 10-mer sequences, which bind 1 protein dimer/duplex, these 22-mer thiophosphate aptamers bind with a stoichiometry of 2 dimers/duplex.


StcU-2 Gene Mutation via CRISPR/Cas9 Leads to Misregulation of Spore-Cyst Formation in Ascosphaera apis.
Ascosphaera apis is the causative agent of honey bee chalkbrood disease, and spores are the only known source of infections. Interference with sporulation is therefore a promising way to manage A. apis. The versicolorin reductase gene (StcU-2) is a ketoreductase protein related to sporulation and melanin biosynthesis. To study the StcU-2 gene in ascospore production of A. apis, CRISPR/Cas9 was used, and eight hygromycin B antibiotic-resistant transformants incorporating enhanced green fluorescent protein (EGFP) were made and analyzed. PCR amplification, gel electrophoresis, and sequence analysis were used for target gene editing analysis and verification. The CRISPR/Cas9 editing successfully knocked out the StcU-2 gene in A. apis. StcU-2 mutants had shown albino and non-functional spore-cyst development and lost effective sporulation. In conclusion, editing of StcU-2 gene has shown direct relation with sporulation and melanin biosynthesis of A. apis; this effective sporulation reduction would reduce the spread and pathogenicity of A. apis to managed honey bee. To the best of our knowledge, this is the first time CRISPR/Cas9-mediated gene editing has been efficiently performed in A. apis, a fungal honey bee brood pathogen, which offers a comprehensive set of procedural references that contributes to A. apis gene function studies and consequent control of chalkbrood disease.


Differentially expressed microRNAs in chondrocytes from distinct regions of developing human cartilage.
There is compelling in vivo evidence from reports on human genetic mutations and transgenic mice that some microRNAs (miRNAs) play an important functional role in regulating skeletal development and growth. A number of published in vitro studies also point toward a role for miRNAs in controlling chondrocyte gene expression and differentiation. However, information on miRNAs that may regulate a specific phase of chondrocyte differentiation (i.e. production of progenitor, differentiated or hypertrophic chondrocytes) is lacking. To attempt to bridge this knowledge gap, we have investigated miRNA expression patterns in human embryonic cartilage tissue. Specifically, a developmental time point was selected, prior to endochondral ossification in the embryonic limb, to permit analysis of three distinct populations of chondrocytes. The location of chondroprogenitor cells, differentiated chondrocytes and hypertrophic chondrocytes in gestational day 54-56 human embryonic limb tissue sections was confirmed both histologically and by specific collagen expression patterns. Laser capture microdissection was utilized to separate the three chondrocyte populations and a miRNA profiling study was carried out using TaqMan® OpenArray® Human MicroRNA Panels (Applied Biosystems®). Here we report on abundantly expressed miRNAs in human embryonic cartilage tissue and, more importantly, we have identified miRNAs that are significantly differentially expressed between precursor, differentiated and hypertrophic chondrocytes by 2-fold or more. Some of the miRNAs identified in this study have been described in other aspects of cartilage or bone biology, while others have not yet been reported in chondrocytes. Finally, a bioinformatics approach was applied to begin to decipher developmental cellular pathways that may be regulated by groups of differentially expressed miRNAs during distinct stages of chondrogenesis. Data obtained from this work will serve as an important resource of information for the field of cartilage biology and will enhance our understanding of miRNA-driven mechanisms regulating cartilage and endochondral bone development, regeneration and repair.


Neural correlates of focused attention during a brief mindfulness induction.
Mindfulness meditation-the practice of attending to present moment experience and allowing emotions and thoughts to pass without judgment-has shown to be beneficial in clinical populations across diverse outcomes. However, the basic neural mechanisms by which mindfulness operates and relates to everyday outcomes in novices remain unexplored. Focused attention is a common mindfulness induction where practitioners focus on specific physical sensations, typically the breath. The present study explores the neural mechanisms of this common mindfulness induction among novice practitioners. Healthy novice participants completed a brief task with both mindful attention [focused breathing (FB)] and control (unfocused attention) conditions during functional magnetic resonance imaging (fMRI). Relative to the control condition, FB recruited an attention network including parietal and prefrontal structures and trait-level mindfulness during this comparison also correlated with parietal activation. Results suggest that the neural mechanisms of a brief mindfulness induction are related to attention processes in novices and that trait mindfulness positively moderates this activation.


Root standing crop and chemistry after six years of soil warming in a temperate forest.
Examining the responses of root standing crop (biomass and necromass) and chemistry to soil warming is crucial for understanding root dynamics and functioning in the face of global climate change. We assessed the standing crop, total nitrogen (N) and carbon (C) compounds in tree roots and soil net N mineralization over the growing season after 6 years of experimental soil warming in a temperate deciduous forest in 2008. Roots were sorted into four different categories: live and dead fine roots (≤1mm in diameter) and live and dead coarse roots (1-4 mm in diameter). Total root standing crop (live plus dead) in the top 10 cm of soil in the warmed area was 42.5% (378.4 vs. 658.5 g m(-2)) lower than in the control area, while live root standing crop in the warmed area was 62% lower than in the control area. Soil net N mineralization over the growing season increased by 79.4% in the warmed relative to the control area. Soil warming did not significantly change the concentrations of C and C compounds (sugar, starch, hemicellulose, cellulose and lignin) in the four root categories. However, total N concentration in the live fine roots in the warmed area was 10.5% (13.7 vs. 12.4 mg g(-1)) higher and C:N ratio was 8.6% (38.5 vs. 42.1) lower than in the control area. The increase in N concentration in the live fine roots could be attributed to the increase in soil N availability due to soil warming. Net N mineralization was negatively correlated with both live and dead fine roots in the mineral soil that is home to the majority of roots, suggesting that soil warming increases N mineralization, decreases fine root biomass and thus decreases C allocation belowground.


Cooperative photoinduced two-dimensional condensation in Langmuir films observed using nanosecond pump-probe Brewster angle microscopy.
Two-dimensional condensation was initiated in a self-assembled mixed monolayer of spiropyran and octadecanol by a nanosecond laser pulse. The dynamics of the process were monitored using nanosecond pump-probe Brewster angle microscopy. Domain growth followed a power law with a growth exponent of 0.47 at a velocity approaching 20 ms(-1). This represents a limit for the rate of longitudinal signaling of pressure waves through a self-assembled amphiphilic layer at an interface and adds to our understanding of signal transmission rates in biomimetic membranes where morphological change in one region can be signaled to a more remote region.


Caffeic acid phenethyl ester reversed cadmium-induced cell death in hippocampus and cortex and subsequent cognitive disorders in mice: Involvements of AMPK/SIRT1 pathway and amyloid-tau-neuroinflammation axis.
Exposure to nonbiodegradable cadmium (Cd) causes many health problems including the damage to the nervous system. This study aimed to increase knowledge about its neurotoxic effects and the neuroprotective potential of caffeic acid phenethyl ester (CAPE, a polyphenol abundant in honeybee propolis). In mice, CAPE (10 μmol/kg/day body weight) attenuated significantly learning and memory deficits induced by CdCl2 (1.5 mg/kg/day body weight). For the CdCl2-treated mice, CAPE increased crossing number in open field test, decreased the alternation in Y-maze test, and increased the latency time and error number in step down test. CAPE also inhibited CdCl2-initiated Aβ accumulation and activation of pro-inflammatory factors and microglia in the brains. Therefore, CAPE could be a food-derived neuroprotective agent against Cd-induced neurotoxicity and neurodegenerative disorders, through attenuating neuronal apoptosis and neuroinflammation via the AMPK/SIRT1 pathway and amyloid-tau-neuroinflammation axis.


Limited gene flow and pronounced population genetic structure of Eastern Massasauga (Sistrurus catenatus) in a Midwestern prairie remnant.
As anthropogenic changes continue to ecologically stress wildlife, obtaining measures of gene flow and genetic diversity are crucial for evaluating population trends and considering management and conservation strategies for small, imperiled populations. In our study, we conducted a molecular assessment to expand on previous work to elucidate patterns of diversity and connectivity in the remaining disjunct Eastern Massasauga Rattlesnake (Sistrurus catenatus) hibernacula in Illinois. We assayed genetic data for 327 samples collected during 1999-2015 from the Carlyle Lake study area across 21 microsatellite loci. We found hibernacula formed distinct genetic clusters corresponding to the three main study areas (Dam Recreation Areas, Eldon Hazlet State Park, and South Shore State Park). Genetic structuring and low estimates of dispersal indicated that connectivity among these study areas is limited and each is demographically independent. Hibernacula exhibited moderate levels of heterozygosity (0.60-0.73), but estimates of effective population size (5.2-41.0) were low and track census sizes generated via long-term mark-recapture data. Hibernacula at Carlyle Lake, which represent the only Eastern Massasauga remaining in Illinois, are vulnerable to future loss of genetic diversity through lack of gene flow as well as demographic and environmental stochastic processes. Our work highlights the need to include population-level genetic data in recovery planning and suggests that recovery efforts should focus on managing the three major study areas as separate conservation units in order to preserve and maintain long-term adaptive potential of these populations. Specific management goals should include improving connectivity among hibernacula, maintaining existing wet grassland habitat, and minimizing anthropogenic sources of mortality caused by habitat management (e.g., mowing, prescribed fire) and recreational activities. Our molecular study provides additional details about demographic parameters and connectivity at Carlyle Lake that can be used to guide recovery of Eastern Massasauga in Illinois and throughout its range.


Antimicrobial activity of antidepressants on normal gut microbiota: Results of the in vitro study.
Currently, there is little published data on the effects of antidepressants on normal gut microbiota and the consequences of such effects on treatment outcomes. The aim of the study: was to evaluate the growth kinetics of normal human gut microorganisms with antidepressants most common in routine clinical practice. Materials and methods: Research objects were species of microorganisms representing normal gut microbiota: Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Candida albicans ATCC 24433, Bifidobacterium 791, Enterococcus faecalis ATCC 29212, Lactobacillus rhamnosus ATCC 53103. All microorganisms were cultivated in Schaedler broth (HiMedia) under aerobic/anaerobic conditions. The active substances of all studied antidepressants (fluvoxamine, fluoxetine, escitalopram, duloxetine, venlafaxine, mirtazapine) were extracted from ground preparations by dimethyl sulfoxide and centrifuged. Each solution of antidepressants was added to a Schaedler broth containing a certain microorganism's strain and diluted to final concentrations-200 μg/ml, 500 μg/ml, and 700 μg/ml. For a quantitative assessment of the effect, the specific growth rates (μ, h-1) of microorganisms were calculated as the slope of the initial part of the growth curve in coordinates (lnA, t). To evaluate the antidepressant effects on representatives of the normal microbiota in vitro, the following parameters were chosen: specific growth rate and IC50. Results: All antidepressants had an inhibitory effect on the growth of all studied microorganisms. Fluvoxamine and venlafaxine had the least effect on the growth activity of all studied microorganisms. Fluoxetine showed a pronounced effect on growth activity against E. coli, E. feacalis, S. aureus, and the least effect against C. albicans. Escitalopram had a greater effect on the growth rate of E. coli, E. feacalis, B. bifidum, L. rhamnosus, and C. albicans, which puts it among the leaders in terms of its effect on the growth activity of the microorganisms we studied. Mirtazapine, according to the results of our experiment, showed the greatest activity against L. rhamnosus and C. albicans. Conclusions: Our results confirm the effects of antidepressants on the growth activity of the normal gut microbiota individual strains. Further study of the antimicrobial activity of antidepressants may become one of the new directions for optimizing the personalized therapy of patients with depression.


Evidence that mnesic states govern normal and disordered memory.
Although it is recognized that retrieval may be state-dependent, only recently has a paradigm been identified that allows state-dependence in rats to be demonstrated reliably and at relevant doses of CNS agents. In humans, the effects of scopolamine constitute a valuable model of disordered memory. We used this paradigm to analyze the effects of scopolamine on different memory processes. Rats treated with either saline or 0.01-10 mg/kg doses of scopolamine learned to lever press for milk reward, and were then tested for retrieval while given the same or a different treatment. Saline-to-scopolamine as well as scopolamine-to-saline state changes produced robust failures to retrieve the response. Remarkably, the state produced by 2.5 mg/kg scopolamine, like that produced by saline, produced little intrinsic effect on learning or any other memory process (i.e. when the prevailing state was left unchanged). However, changing the implemented state from one to the other between two different processes, disrupted not only retrieval, but also learning, encoding and retention. We also determined whether the graded state changes produced by 0.01-10 mg/kg doses of scopolamine could mimic the peculiar and poorly understood temporally graded retrograde amnesia that occurs in Alzheimer's disease. In rats that had acquired a complex drug-discrimination task over a 6-month period, scopolamine-induced state changes seemed to produce dose-dependent deficits in the retrieval of recent information while preserving those abilities that had been acquired in the increasingly remote past. Beyond its role in retrieval, the findings implicate state dependence in learning, encoding and retention, and suggest that physiologically defined mnesic states govern each of these. The changes of mnesic state that are likely associated with excessively labile cholinergic neurotransmission may conceivably cause the complex disabilities of Alzheimer's disease.


Quantifying potential N turnover rates in hypersaline microbial mats by 15N tracer techniques.
Microbial mats, due to stratification of the redox zones, have a potential to include a complete N cycle, however an attempt to evaluate a complete N cycle in these ecosystems has not been yet made. In this study, occurrence and rates of major N cycle processes were evaluated in intact microbial mats from Elkhorn Slough, Monterey Bay, CA, USA, and Baja California Sur, Mexico under oxic and anoxic conditions using 15N-labeling techniques. All of the major N transformation pathways, with the exception of anammox, were detected in both microbial mats. Nitrification rates were found to be low at both sites for both seasons investigated. The highest rates of ammonium assimilation were measured in Elkhorn Slough mats in April and corresponded to high in situ ammonium concentration in the overlying water. Baja mats featured higher ammonification than ammonium assimilation rates and this, along with their higher affinity for nitrate compared to ammonium and low dissimilatory nitrate reduction to ammonium rates, characterized their differences from Elkhorn Slough mats. Nitrogen fixation rates in Elkhorn Slough microbial mats were found to be low implying that other processes such as recycling and assimilation from water are main sources of N for these mats at the times sampled. Denitrification in all of the mats was incomplete with nitrous oxide as end product and not dinitrogen. Our findings highlight N cycling features not previously quantified in microbial mats and indicate a need of further investigations in these microbial ecosystems.Importance: Nitrogen is essential for life. The nitrogen cycle on Earth is mediated by microbial activity and has had a profound impact on both the atmosphere and the biosphere throughout geologic time. Microbial mats, present in many modern environments, have been regarded as living records of the organisms, genes, and phylogenies of microbes, as they are one of the most ancient ecosystems on Earth. While rates of major nitrogen metabolic pathways have been evaluated in a number of ecosystems, it remains elusive in microbial mats. In particular it is unclear what factors affect nitrogen cycling in these ecosystems and how morphological differences between mats impact nitrogen transformations. In this study we investigate nitrogen cycling in two microbial mats having morphological differences. Our findings provide insight for further understanding of biogeochemistry and microbial ecology of microbial mats.


Carotid intima-media thickness and risk of cardiovascular events in high-risk patients. Results of the Osaka Follow-Up Study for Carotid Atherosclerosis 2 (OSACA2 Study).
There is epidemiological evidence that increased carotid intima-media thickness (IMT) is a predictor of cardiovascular disease (CVD) events. However, the significance of carotid IMT in high-risk patients in whom risk factors are managed clinically has not been adequately investigated. The purpose of this study was to determine the usefulness of carotid IMT measurement in such patients. The study comprised 900 outpatients with cardiovascular risk factors or established atherosclerosis. Carotid IMT was calculated as the mean bilateral IMT of the common carotid artery, bifurcation, and internal carotid artery. Baseline vascular risk factors, medications, and history of CVD were recorded at the time of enrollment. The incidence of CVD events was determined prospectively. During a mean follow-up period of 2.6 years, there were 64 CVD events. The relative risk (RR) of a CVD event increased with increased IMT. Association between CVD events and carotid IMT was significant after adjustment for risk factors and history of CVD, showing an increased risk per IMT tertile from the middle tertile (RR, 2.5; 95% confidence interval [CI]: 1.0-6.3) to the highest (RR, 3.6; 95% CI: 1.4-9.0). When patients with a history of CVD were excluded (n = 574), the predictive value of IMT was significant even after adjustment for risk factors (hazard ratio per 1 SD IMT increase was 1.57 [95% CI: 1.11-2.20]). Carotid IMT is an independent predictor of vascular events in high-risk patients in whom risk factors are managed clinically.


Posttranslational Protein Modifications in Type 1 Diabetes - Genetic Studies with PCMT1, the Repair Enzyme Protein Isoaspartate Methyltransferase (PIMT) Encoding Gene.
Posttranslational protein modifications have been implicated in the development of autoimmunity. Protein L-isoaspartate (D-aspartate) O-methyltransferase (PIMT) repairs modified proteins and is encoded by PCMT1, located in a region linked to type 1 diabetes (T1D), namely IDDM5. To evaluate the association between genetic variations in the PCMT1 gene and T1D. Firstly, PCMT1 was sequenced in 26 patients with T1D (linked to IDDM5) and 10 control subjects. The variations found in PCMT1 were then tested (alone and interacting with a functional polymorphism in SUMO4 and with HLA) for association with T1D in 253 families (using transmission disequilibrium test). In a third step, the association of the functional variation in PCMT1 (rs4816) with T1D was analyzed in 778 T1D patients and 749 controls (using chi-square test). In vitro promoter activity was assessed by transfecting INS-1E cells with PCMT1 promoter constructs and a reporter gene, with or without cytokine stimulation. Four polymorphisms in complete linkage disequilibrium were identified in PCMT1 (5' to the gene (rs11155676), exon 5 (rs4816) and exon 8 (rs7818 and rs4552)). In the whole cohort of 253 families, the allele associated with increased PIMT enzyme activity (rs4816, allele A) was less frequently transmitted to the affected than to the non-affected offspring (46% vs. 53%, p = 0.099). This finding was even more evident in the subset of families where the proband had high-risk SUMO4 (p = 0.069) or low-risk HLA (p = 0.086). Surprisingly, in the case-control study with 778 cases and 749 controls, an inverse trend was found (40.36% of patients and 36.98% of controls had the allele, p = 0.055). PCMT1 promoter activity increased with cytokine stimulation, but no differences were detected between the constructs adjacent to rs11155676. PCMT1 was virtually associated with T1D in groups defined by other risk genes (SUMO4 and HLA). A general association in a not further defined sample of T1D patients was not evident. Verification in a larger population is needed.


Is the AO spine thoracolumbar injury classification system reliable and practical? a systematic review.
Controversy surrounding the classification of thoracolumbar injuries has given rise to various classification systems over the years, including the most recent AOSpine Thoracolumbar Injury Classification System (ATLICS). This systematic review aims to provide an up-to-date evaluation of the literature, including assessment of a further three studies not analysed in previous reviews. In doing so, this is the first systematic review to include the reliability among non-spine subspecialty professionals and to document the wide variety between reliability across studies, particularly with regard to sub-type classification. Relevant studies were found via a systematic search of PubMed, EBESCO, Cochrane and Web of Science. Data extraction and quality assessment were conducted in line with Cochrane Collaboration guidelines. Twelve articles assessing the reliability of ATLICS were included in this review. The overall inter-observer reliability varied from fair to substantial, but the three additional studies in this review, compared to previous reviews, presented on average only fair reliability. The greatest variation of results was seen in A1 and B3 subtypes. Least reliably classified on average was A4 subtype. This systematic review concludes that ATLICS is reliable for the majority of injuries, but the variability within subtypes suggests the need for further research in assessing the needs of users in order to increase familiarity with ATLICS or perhaps the necessity to include more subtype-specific criteria into the system. Further research is also recommended on the reliability of modifiers, neurological classification and the application of ATLICS in a paediatric context.


Psychological functioning and bodily conditions in patients with pain disorder associated with psychological factors.
A sample of chronic pain patients (N = 40) was described with reference to defence mechanisms, interpersonal problems, psychological symptoms and bodily conditions. The relationships between pain intensity and different psychological and bodily indexes were examined. The defence mechanisms of somatization and denial measured by the Minnesota Multiphasic Personality Inventory (MMPI) characterized the sample. Interpersonal relations were typically overly nurturant, exploitable, non-assertive and socially avoidant according to the circumplex version of Inventory of Interpersonal Problems (IIP-C). Somatization, obsession, depression and anxiety were the highest symptom scales on the Symptom Check-List 90, revised (SCL-90-R). The Comprehensive Body Examination (CBE) produced moderate findings mainly reflecting stiffness, and the intensity of pain was medium high. The MMPI psychosomatic pattern, the combined IIP-C index consisting of the elevated subscales, and the elevated subscales on SCL-90-R were all moderately correlated with pain intensity. Contrary to our prediction, the global bodily stiffness score was unrelated to pain.


Increased brainstem angiotensin-I levels in chronic 2 KIC hypertensive rats.
A study was undertaken on the role of brainstem renin-angiotensin system in maintenance of hypertension in chronic renovascular hypertensive rats. Hypertension was induced by unilateral renal artery clamping, while the contralateral kidney was left intact (2 KIC). Blood pressure (BP), plasma renin activity (PRA) and brainstem angiotensin (ang-I) levels were measured after 24 days, in hypertensive and sham-operated animals. In separate subgroups of these animals, the effect of intracerebroventricular administration of captopril on the parameters listed was studied. The results showed high ang-I levels in 2 KIC rats as compared to controls (P less than 0.05). Captopril administration (500 micrograms/50 microliters icv) caused a fall in BP and increase in brainstem ang-I levels (P less than 0.01). In control animals, however, captopril produced a rise in BP without any significant change in brainstem ang-I levels. Peripheral plasma renin activity was normal, despite significant sodium retention in 2 KIC rats. The results are suggestive of activation of brainstem renin-angiotensin system (RAS) in chronic 2 KIC hypertension.


[Familial adenomatous polyposis: establishing a registry and genetic and molecular analysis].
Familial adenomatous polyposis (FAP), a dominantly inherited disease, is caused by a mutation in the adenomatous polyposis coli gene in chromosome 5q21. The gene has 15 exons, a physical length of 10 Kb and an open reading frame of 8.5 Kb. Exon 15 codes 66% of the mRNA and has a mutation cluster region which accounts for over 50% of mutations. The disease usually leads to the appearance of hundreds of adenomatous polyps in the transverse and descending colon between puberty and age 20 years and to colon cancer before the age of 40. Early detection is essential to prevent the development of metastasizing cancer. Since 1994 we have recruited 23 families for genetic counseling. DNA was obtained from 19 unrelated FAP patients and 219 high risk relatives in 19 unrelated families following confirmation of the diagnosis. In addition to linkage studies, direct mutational analysis was performed using the protein truncation test for most of exon 15 and single strand conformation polymorphism analysis for the other exons. These exons account for most of the mutations identified to date. Of 19 unrelated probands, 14 had detectable mutations. Exon 15 accounted for 6 families, exons 5, 7 and 14 for 1 each, exon 9 for 3, and exon 8 for 2. Combined mutational and linkage analysis identified 18 presymptomatic carriers who received genetic and clinical counseling. Our FAP patients did not differ significantly from those of larger studies in other countries with regard to the distribution of the mutations, gender and genotype-phenotype correlation, or ethnic distribution.


Segmentation of psoriasis vulgaris images using multiresolution-based orthogonal subspace techniques.
In this paper, a method is proposed for the segmentation of color images using a multiresolution-based signature subspace classifier (MSSC) with application to psoriasis images. The essential techniques consist of feature extraction and image segmentation (classification) methods. In this approach, the fuzzy texture spectrum and the two-dimensional fuzzy color histogram in the hue-saturation space are first adopted as the feature vector to locate homogeneous regions in the image. Then these regions are used to compute the signature matrices for the orthogonal subspace classifier to obtain a more accurate segmentation. To reduce the computational requirement, the MSSC has been developed. In the experiments, the method is quantitatively evaluated by using a similarity function and compared with the well-known LS-SVM method. The results show that the proposed algorithm can effectively segment psoriasis images. The proposed approach can also be applied to general color texture segmentation applications.


Peer support in anesthesia: turning war stories into wellness.
Peer support, a method of providing for the well being of healthcare providers following adverse or stressful events, is garnering increased attention in light of the increased prevalence and awareness of burnout, depression and suicidality in physicians. In this review, we will summarize the evolution of the 'second victim,' explore methods of support and examine how new regulatory requirements are changing the peer support landscape. As peer support and the second victim are investigated more, themes are emerging regarding the natural history of recovery. As these are delineated, more targeted peer support models are being developed. One major change in 2017 is the institution of new Accreditation Council for Graduate Medical Education's Common Program Requirements, now including topics targeted on well being. Effective and accessible peer support is developing in many departments nationwide and can only be expected to continue, given new regulatory requirements. As these programs develop, and research on their effect continues, best practices will likely emerge.


Life cycle of the camel tick Hyalomma dromedarii (Acari: Ixodidae) under field conditions in Northern Sudan.
Engorged Hyalomma dromedarii females were placed for development in shade and sun (open non-shaded site) in April, August and December for two successive years. Engorged nymphs were also placed at the same sites in January, March, May, June, July and August. Preoviposition periods ranged between 9.8 and 11.7 days in the shade but longer in the sun in December (14.7 days). Egg production index was higher in August (0.72) than in April and December (0.46 and 0.39, respectively) in shade and in August compared to December in the sun (0.65 and 0.29, respectively). In shade and in sun, the prehatching periods were longer in December (50.7 and 48.2 days) than in April and August (25.6 and 29.0 days). Percentage hatchability ranged between 82 to 94.1% and 56.2 and 58.9% in the shade and sun, respectively. Number of eggs laid and eggs hatched were positively correlated with females' engorgement weights. There were high mortality rates of females in the sun and most of the eggs desiccated. Slight increased ambient and soil temperatures increased egg production, shortened preoviposition periods and decreased hatchability while increased humidity shortened preoviposition and prehatching periods and increased hatchability. Nymphal-adult moulting periods were long in January (20.7 days in the shade and 14.4 days in the sun) and short in August in the shade (7.8 days) while they were only 6 days in May and June in the sun. High numbers moulted in all months in the shade (ranges 23.3-29.5 nymphs). The mean mortality of engorged nymphs in the shade was very low. However, in the sun mortality was high (12.9-30 nymphs). Premoulting periods were negatively correlated with ambient temperature but positively correlated with humidity. In the shade, mean number of nymphs moulting increased with increased mean ambient temperature, humidity and rainfall.


Metastasis of melanoma to the gastrointestinal tract. Report of three cases
Cutaneous melanoma is one of the most common malignancies to metastasize to the gastrointestinal tract. Presence of tumor metastasis is an important prognostic factor. Despite clinical advances, the five-year survival rate of patients with stage IV malignant melanoma is only about 14%. We present three clinical cases with metastasis of cutaneous melanoma. Man 68 years old and woman 55 years old, with colon metastasis; and man 75 years old with metastasis to the small intestine. All of them underwent resection alone when they presented gastrointestinal symptoms and they obtained relief after it. Two patients had initial cutaneous lesions with Breslow´s thickness < 2 mm. The three patients died months after the surgery. Meticulous attention to patients with prior history of cutaneous melanoma who present with subtle gastrointestinal symptoms is highly recommended.


Para-Cardiac Inflammatory Mass Compressing the Heart: A possible association with COVID-19.
Infection with the SARS-CoV-2 virus causes coronavirus disease 2019 (COVID-19). COVID-19 usually affects the lungs but may also involve other organs such as the heart. We report a case of a para-cardiac mass in a previously healthy 45-year-old male who developed persistent dyspnea following SARS-CoV-2 infection. The patient underwent cardiac surgery since the mass was attached to the pericardium and was causing constrictive pericarditis. The pathology report indicated an inflammatory pattern for the mass. Based on the authors' knowledge there has been no previous report of developing a para-cardiac inflammatory mass after SARS-CoV-2 infection. This report aimed to increase awareness regarding the possibility of developing a para-cardiac inflammatory mass following COVID-19.


Muscle function in glenohumeral joint stability during lifting task.
Ensuring glenohumeral stability during repetitive lifting tasks is a key factor to reduce the risk of shoulder injuries. Nevertheless, the literature reveals some lack concerning the assessment of the muscles that ensure glenohumeral stability during specific lifting tasks. Therefore, the purpose of this study was to assess the stabilization function of shoulder muscles during a lifting task. Kinematics and muscle electromyograms (n = 9) were recorded from 13 healthy adults during a bi-manual lifting task performed from the hip to the shoulder level. A generic upper-limb OpenSim model was implemented to simulate glenohumeral stability and instability by performing static optimizations with and without glenohumeral stability constraints. This procedure enabled to compute the level of shoulder muscle activity and forces in the two conditions. Without the stability constraint, the simulated movement was unstable during 74%±16% of the time. The force of the supraspinatus was significantly increased of 107% (p<0.002) when the glenohumeral stability constraint was implemented. The increased supraspinatus force led to greater compressive force (p<0.001) and smaller shear force (p<0.001), which contributed to improved glenohumeral stability. It was concluded that the supraspinatus may be the main contributor to glenohumeral stability during lifting task.


Identification and characterization of a thermostable MutS homolog from Thermus aquaticus.
Recognition of mispaired or unpaired bases during DNA mismatch repair is carried out by the MutS protein family. Here, we describe the isolation and characterization of a thermostable MutS homolog from Thermus aquaticus YT-1. Sequencing of the mutS gene predicts an 89.3-kDa polypeptide sharing extensive amino acid sequence homology with MutS homologs from both prokaryotes and eukaryotes. Expression of the T. aquaticus mutS gene in Escherichia coli results in a dominant mutator phenotype. Initial biochemical characterization of the thermostable MutS protein, which was purified to apparent homogeneity, reveals two thermostable activities, an ATP hydrolysis activity in which ATP is hydrolyzed to ADP and Pi and a specific DNA mismatch binding activity with affinities for heteroduplex DNAs containing either an insertion/deletion of one base or a GT mismatch. The ATPase activity exhibits a temperature optimum of approximately 80 degrees C. Heteroduplex DNA binding by the T. aquaticus MutS protein requires Mg2+ and occurs over a broad temperature range from 0 degrees C to at least 70 degrees C. The thermostable MutS protein may be useful for further biochemical and structural studies of mismatch binding and for applications involving mutation detection.


Efficacy and complications of argon plasma coagulation for hematochezia related to radiation proctopathy.
Endoscopic treatments effectively control bleeding caused by radiation proctopathy. The aims of this study were to determine the efficacy and side effects of argon plasma coagulation in the treatment of this type of bleeding. Records of 21 consecutive patients in whom argon plasma coagulation was used to treat hemorrhagic radiation proctopathy were reviewed. Pharmacologic measures had been unsuccessful in 12 patients. Endoscopic treatment had been unsuccessful in 5 patients. All patients were anemic and 4 had received blood transfusions. The mean number of treatment sessions was 1.7, and 10 patients were successfully treated in single session. Rectal bleeding resolved within 1 month of the last treatment in 19 patients, usually on the day of the last procedure. Bleeding resolved 2 months after cessation of therapy in another patient. Short-term side effects occurred in 3 (14%) patients (rectal pain, tenesmus, and/or abdominal distention); long-term complications (rectal pain, tenesmus, diarrhea) developed in 4 patients (19%). Hematochezia caused by radiation proctopathy is effectively controlled by argon plasma coagulation, in some cases after a single treatment session. Treatment may result in protracted bowel symptoms.


The Localization of Cytochrome P450s CYP1A1 and CYP1A2 into Different Lipid Microdomains Is Governed by Their N-terminal and Internal Protein Regions.
In cellular membranes, different lipid species are heterogeneously distributed forming domains with different characteristics. Ordered domains are tightly packed with cholesterol, sphingomyelin, and saturated fatty acids, whereas disordered domains contain high levels of unsaturated fatty acids. Our laboratory has shown that membrane heterogeneity affects the organization of cytochrome P450s and their cognate redox partner, the cytochrome P450 reductase (CPR). Despite the high degree of sequence similarity, CYP1A1 was found to localize to disordered regions, whereas CYP1A2 resided in ordered domains. We hypothesized that regions of amino acid sequence variability may contain signal motifs that direct CYP1A proteins into ordered or disordered domains. Thus, chimeric constructs of CYP1A1 and CYP1A2 were created, and their localization was tested in HEK293T cells. CYP1A2, containing the N-terminal regions from CYP1A1, no longer localized in ordered domains, whereas the N terminus of CYP1A2 partially directed CYP1A1 into ordered regions. In addition, intact CYP1A2 containing a 206-302-residue peptide segment of CYP1A1 had less affinity to bind to ordered microdomains. After expression, the catalytic activity of CYP1A2 was higher than that of the CYP1A1-CYP1A2 chimera containing the N-terminal end of CYP1A1 with subsaturating CPR concentrations, but it was approximately equal with excess CPR suggesting that the localization of the CYP1A enzyme in ordered domains favored its interaction with CPR. These data demonstrate that both the N-terminal end and an internal region of CYP1A2 play roles in targeting CYP1A2 to ordered domains, and domain localization may influence P450 function under conditions that resemble those found in vivo.


Mechanism of regulation of amylase release by alpha- and beta-adrenergic agonists in rat parotid tissue.
The effect of forskolin and isobutyl-methylxanthine on amylase release and cyclic AMP accumulation by alpha- and beta-adrenergic agonists was studied in rat parotid slices. A small increase in cyclic AMP by beta-adrenergic agonists was sufficient for maximum stimulation of amylase release (Yoshimura et al., 1982a), but a similar increase in cyclic AMP by forskolin did not stimulate the maximum amount of amylase release. The amount of amylase release and cyclic AMP changed in parallel when lower doses of isobutyl-methylxanthine were used, but higher doses of isobutyl-methylxanthine further increased cyclic AMP without causing a significant increase in amylase release. Amylase release stimulated to its maximum by isobutyl-methylxanthine was much lower than that by isoproterenol. These results suggest that cyclic AMP is not the only chemical correlate for the activation of amylase release by beta-adrenergic agonists. The effect of phenylephrine and methoxamine on amylase release was augmented by either forskolin or isobutyl-methylxanthine, but the effect of methacholine was not. Phenylephrine increased the cyclic AMP concentration under the same conditions, but methoxamine did not. The inhibition of the effect of phenylephrine plus forskolin on cyclic AMP accumulation by propranolol was almost complete and stereospecific, but the inhibition of their effects on amylase release was incomplete and not stereospecific. Synergism of amylase release was observed in the effect between methoxamine and dibutyryl-cyclic AMP. These results suggest that the augmentation of the effect of alpha-adrenergic agonists on amylase release by forskolin or isobutyl-methylxanthine cannot be explained only on changes in cyclic AMP and that some other factor in collaboration with cyclic AMP may participate in the regulation of amylase release by alpha-adrenergic agonists.


Can a Portable Balance System Predict Fall Risk in Community-Dwelling Older Women?
Increased fall risk has been linked to age, being female, and age-related changes in the postural control system. The purpose of this study was to determine if a portable balance system could predict fall risk by determining the relationship among the modified Clinical Test for Sensory Interaction on Balance (mCTSIB) scores, age, the Activities-specific Balance Confidence Scale (ABC) score, and the Berg Balance Scale (BBS) score in community-dwelling older women. Insight into these relationships may facilitate early intervention and decrease fall risk in older women. This study was a non-experimental, prospective, cross-sectional, exploratory analysis to determine the relationship among the mCTSIB, age, ABC, and BBS. Women aged 65 years and over were selected from two independent living facilities using a sample of convenience (n=42). The mCTSIB firm surface, eyes open and the ABC predicted fall risk (BBS) scores, F(2, 36)=35.72, p<0.001, R2=0.67, but adding age did not significantly improve the model, b= -0.17, t(36)= -1.71, p=0.10. A portable balance system may be an effective screening tool to predict fall risk in community-dwelling older women and may be used by a variety of allied health professionals. Postural sway, and perceived balance, predicted fall risk scores (BBS). More specifically, data obtained from the mCTSIB firm surface, eyes open test condition when combined with ABC scores could lead to identification of increased fall risk, allowing clinicians to recommend early treatment intervention to prevent future falls.


Relationship between growth behaviour, micro and macroscopic morphologies and freezing sensitivity of the ripening starter Geotrichum candidum is strain specific and mostly related to the morphotypes: the arthrospores/hyphae parameter.
Microscopic conformation, growth behaviour and freezing sensitivity of seven strains of Geotrichum candidum, a ripening starter, were studied and compared according to their macroscopic morphotypes. It has been shown that the thallus forming units (TFU)×ml-1/OD600nm ratio as a function of time is an interesting parameter to follow G. candidum sporulation through the growth behaviour. Microscopic conformation, growth behaviour and freezing sensitivity are clearly strain specific and mostly related to their corresponding morphotypes "yeast", "mould" or "intermediate". The two "mould" strains that sporulate weakly (UCMA103, UCMA499) showed a low survival rate to freezing stress whereas the "yeast" strains expressed a significant resistance owing to the arthrospore abundance. Interestingly, one strain (UCMA96) which appeared on solid medium in accord with the "mould" morphotype respond similarly to freezing stress.


Efficacy and safety of ritonavir-boosted dual protease inhibitor therapy in antiretroviral-naive HIV-1-infected patients: the 2IP ANRS 127 study.
We evaluate the efficacy and tolerability of ritonavir-boosted dual protease inhibitor as a nucleoside reverse transcriptase inhibitor-sparing regimen in a prospective open-label randomized pilot trial in antiretroviral-naive patients. Thirty patients received fosamprenavir/atazanavir/ritonavir (Group 1) and 31 patients received saquinavir/atazanavir/ritonavir (Group 2). The primary endpoint for efficacy was the rate of early virological success, defined as plasma viral load <50 copies/mL at week 16. The study is registered with ClinicalTrials.gov (NCT00122603). At baseline, median (range) viral load was 4.8 log(10) copies/mL (4.0-5.7) and the median CD4 cell count was 271/mm(3) (197-740). Viral load was <50 copies/mL in 12/30 patients [40%, 95% confidence interval (CI) 23%-58%] and 13/31 patients (42%, 95% CI 25%-59%) at week 16 in Groups 1 and 2, respectively. Patients with failing regimens (viral load >or=400 copies/mL at week 16 or >or=50 copies/mL at week 24) were switched to a standard antiretroviral regimen. At week 48, by an intention-to-treat analysis, 23/30 patients (77%) and 26/31 patients (84%) had plasma HIV-1 RNA <50 copies/mL in Groups 1 and 2, respectively. Four patients discontinued treatment for adverse events, all before week 4. No major changes in the protease gene were detected at treatment failure relative to baseline. Baseline viral load <50 000 copies/mL was the only predictor of virological success at week 16. Ritonavir-boosted dual protease inhibitor regimens targeting only one step of viral replication were insufficient to rapidly suppress plasma HIV RNA to <50 copies/mL in antiretroviral-naive patients with high viral load at baseline.


Evidence-based recommendations for the use of antiemetics in radiotherapy.
To report recommendations given in the Multinational Association of Supportive Care in Cancer (MASCC) International Consensus Conference regarding the use of antiemetics in radiotherapy. A steering committee under MASCC auspice chose panel participants for the guidelines development process on prevention of chemotherapy- and radiotherapy-induced emesis (RIE). Pertinent information from published literature as of March 2004 was reviewed for the guideline process. Both the MASCC level of scientific confidence and level of consensus, and the American Society of Clinical Oncology (ASCO) type of evidence and grade for recommendation were adopted. Total body irradiation is classified at high risk, upper abdomen at moderate, lower thorax, pelvis, cranium (radiosurgery) and craniospinal at low, head and neck, extremities, cranium and breast at minimal risk. The recommendations for the use of antiemetics in radiotherapy are as follows: prophylaxis with a 5-HT3 antagonist in patients at high and moderate risk levels of RIE (+/-dexamethasone in the former group), prophylaxis or rescue with a 5-HT3 antagonist in the low risk group, and rescue with dopamine or a 5-HT3 receptor antagonist in minimal risk level. These recommendations represent a valid tool for prophylaxis and treatment of RIE in clinical practice.


Characterization of Mycobacterium tuberculosis var. africanum isolated from a patient with pulmonary tuberculosis in Brazil.
Human tuberculosis (TB) is caused by members of the Mycobacterium tuberculosis complex (MTBC), including Mycobacterium tuberculosis var. tuberculosis (MTB) and Mycobacterium tuberculosis var. africanum (MAF). While MTB is isolated worldwide, MAF is almost completely restricted to the African continent, and despite the historical proximity between Brazil and Africa during the slave trade, no case of TB being caused by MAF has been reported in Brazil to date. We hereby describe the first case of TB caused by MAF in Brazil comparing its genome against the published ones. A female patient who had never visited Africa presented with clinical symptoms typical of pulmonary TB. Based on 16S rRNA gene sequencing, the cultured isolate was identified as belonging to MTBC and partial sequence of the hsp65 gene was identical to that of MAF. This was confirmed by genotyping based on detection of Single Nucleotide Polymorphism (SNP), Region of Difference (RD) and spoligotyping. The isolate presented the Shared International Typing (SIT) 181. In the whole-genome comparison against MAF genomes available on published EMBL-EBI European Nucleotide Archive (ENA), the Brazilian genome (MAFBRA00707) was identified as belonging to Lineage 6 and clustered with isolates from The Gambia. This is the first report of the isolation of MAF from a patient from Brazil, without evidence of having any contact with an African index case.


Chlorella vulgaris Ameliorates Oxidative Stress and Improves the Muscle Regenerative Capacity of Young and Old Sprague-Dawley Rats.
Muscle atrophy in ageing is a multifactorial degenerative process impacted by cellular ageing biology, which includes oxidative stress. Chlorella vulgaris is a coccoid green eukaryotic microalga rich in antioxidants. The aim of this study was to determine the effect of C. vulgaris in ameliorating oxidative stress, thus elucidating its mechanism in improving muscle mass, strength and function in young and old rats. Fifty-six male Sprague-Dawley (SD) rats aged 3 months (young) and 21 months (old) were divided into three groups: Group 1 (control) was given distilled water; Group 2 was treated with 150 mg/kg body weight (BW) of C. vulgaris; and Group 3 was treated with 300 mg/kg BW of C. vulgaris for three months. Grip and muscle strength and muscle integrity were determined on days 0, 30, 60, and 90 of treatment. Urine and blood were collected on days 0 and 90 of treatment for oxidative stress marker determination, while the gastrocnemius muscles were collected for muscle oxidative stress analysis. Increased grip strength of the front and hind paws was observed in young C. vulgaris-treated rats on days 30, 60, and 90 compared to the untreated control on the same days (p < 0.05). There was a significant increase in lean bone mineral content (BMC) in young rats treated with 300 mg/kg BW C. vulgaris compared to untreated rats on days 30 and 60. The fat mass was significantly decreased in young and old C. vulgaris-treated rats on day 90 compared to the untreated control. The total path was significantly increased for old rats treated with 300 mg/kg BW C. vulgaris on days 60 and 90 compared to day 0. Young and old C. vulgaris-treated rats demonstrated a significant decrease in urinary isoprostane F2t and plasma creatine kinase-MM (CKMM) compared to the control on day 90. A significant decrease in malondialdehyde (MDA) and 4-hydroxyalkenal (HAE) levels were observed in young and old rats treated with C. vulgaris. C. vulgaris improved the muscle mass, strength, and function in young and old rats. This effect could be due to its potency in ameliorating oxidative stress in the skeletal muscle of young and old rats.


[Favorable results of early 2nd-stage medication in rheumatoid arthritis].
To compare the effects of treatment with and without 'slow-acting antirheumatic drugs' (SAARDs) for patients with recent-onset rheumatoid arthritis (RA). Open randomized clinical trial. Outpatient clinics of six medical centers, the Netherlands. The study population consisted of 304 consecutive patients with recently diagnosed RA. A therapeutic strategy with delayed introduction of a SAARD was compared with a strategy comprising early start of SAARD treatment. Primary endpoints were functional disability, pain, joint score, erythrocyte sedimentation rate and progression of radiological abnormalities at 12 months. Four of the five endpoints improved statistically significantly more in the early-SAARD group than in the non-SAARD group; radiological abnormalities progressed at an equal rate in the two groups. Adverse reactions remained at an acceptable level and were the same in the two groups. Early introduction of a SAARD was more beneficial than a therapeutic strategy with delayed introduction of a SAARD for patients with recently diagnosed RA.


Regulatory Networks of the T4SS Control: From Host Cell Sensing to the Biogenesis and the Activity during the Infection.
Delivery of effectors, DNA or proteins, that hijack host cell processes to the benefit of bacteria is a mechanism widely used by bacterial pathogens. It is achieved by complex effector injection devices, the secretion systems, among which Type 4 Secretion Systems (T4SSs) play a key role in bacterial virulence of numerous animal and plant pathogens. Considerable progress has recently been made in the structure-function analyses of T4SSs. Nevertheless, the signals and processes that trigger machine assembly and activity during infection, as well as those involved in substrate recognition and transfer, are complex and still poorly understood. In this review, we aim at summarizing the last updates of the knowledge on signaling pathways that regulate the biogenesis and the activity of T4SSs in important bacterial pathogens.


Short-term effects of repetitive arm work and dynamic exercise on glucose metabolism and insulin sensitivity.
To determine whether repetitive arm work, with a large component of static muscle contraction alters glucose metabolism and insulin sensitivity. Euglycemic clamps (2 h) were started in ten healthy individuals 15 min after 37 min periods of: (1) repetitive arm work in a simulated occupational setting; (2) dynamic concentric exercise on a cycle ergometer at 60% of VO(2max) and (3) a resting regime as a control. During the experimental periods, blood samples were collected, blood pressure was measured repeatedly and electrocardiogram (ECG) was recorded continuously. During the clamps, euglycemia was maintained at 5 mmol l(-1) and insulin was infused at 56 mU m(-2) min(-1) for 120 min. The insulin-mediated glucose disposal rate (M-value) for the steady-state period (60-120 min) of the clamp, tended to be lower following arm work than for both cycling and resting regimes. When dividing the steady-state period into 20-min intervals, the insulin sensitivity index (ISI) was significantly lower for arm work compared with the resting control situation between 60-80 min (P = 0.04) and 80-100 min (P = 0.01), respectively. Catecholamines increased significantly for arm work and cycling compared with resting regime. Data from heart rate variability (HRV) measurements indicated significant sympathetic activation during repetitive arm work. The results indicate that repetitive arm work might acutely promote insulin resistance, whereas no such effect on insulin resistance was produced by dynamic concentric exercise.


In vitro modelling of epithelial and stromal interactions in non-malignant and malignant prostates.
To study the effects of stromal epithelial cell interactions on prostate cancer metastasis, we have used primary human prostatic stromal cells derived from malignant and non-malignant tissues and established epithelial cell lines from normal (PNT1a and PNT2-C2) and tumour (PC-3, DU145 and LNCaP) origins. The effects of stromal cells on epithelial cell growth were studied in direct and indirect (using culture inserts) co-culture and by exposure to stromal cell-conditioned medium (assessed by MTT assay). The influence of stromal cells on epithelial cell invasion was measured using matrigel invasion chambers and on epithelial cell motility using time lapse microscopy. Results indicated that epithelial cell line growth was similarly unaffected or inhibited by stromal cells derived from malignant (n = 8) or non-malignant tissue (n = 8). In contrast, PNT2-C2 and PC-3 cells were found to be the least and the most invasive and motile epithelia respectively. Stromal cultures enhanced the invasion of both epithelial cells, but no differences were observed between the use of malignant and non-malignant tissues. All stromal cultures modestly stimulated PNT2-C2 motility but displayed a greater stimulation of PC-3 cell motility, while stromal cells derived from malignant tissue stimulated PNT2-C2 and PC-3 cell motility more than stromal cultures from non-malignant tissues.


Hospital discharge as experienced by family carers of people with dementia: a case for quality improvement.
To explore whether hospital discharge practices meets the needs of the family carer of a person with dementia. Qualitative methodology utilizing semistructured interviews with the primary family carer of a person with dementia after hospital discharge. Purposive sampling methods were used to recruit family carers within 2 months of discharge from hospitals in the state of Victoria, Australia. Twenty-five family carers were interviewed. Key issues that families found problematic include: coordination of discharge planning, the ability of the staff to address family members' needs, poor engagement with family, and perceived lapses in care delivery. Findings suggest that a number of changes to hospital processes and health professionals' practices are needed in order to better meet the transitional needs of family carers of people with dementia. Considerations for practice to improve the quality of hospital discharge preparation for family carers of people with dementia are provided.


RASSF1A Deficiency Enhances RAS-Driven Lung Tumorigenesis.
Mutant K-RAS has been shown to have both tumor-promoting and -suppressing functions, and growing evidence suggests that the RASSF family of tumor suppressors can act as RAS apoptosis and senescence effectors. It has been hypothesized that inactivation of the RASSF1A tumor suppressor facilitates K-RAS-mediated transformation by uncoupling it from apoptotic pathways such as the Hippo pathway. In human lung tumors, combined activation of K-RAS and inactivation of RASSF1A is closely associated with the development of the most aggressive and worst prognosis tumors. Here, we describe the first transgenic mouse model for activation of K-RAS in the lung in a RASSF1A-defective background. RASSF1A deficiency profoundly enhanced the development of K-RAS-driven lung tumors in vivo Analysis of these tumors showed loss of RASSF1A-uncoupled RAS from the proapoptotic Hippo pathway as expected. We also observed an upregulation of AKT and RALGEF signaling in the RASSF1A- tumors. Heterozygosity of RASSF1A alone mimicked many of the effects of RAS activation on mitogenic signaling in lung tissue, yet no tumors developed, indicating that nonstandard Ras signaling pathways may be playing a key role in tumor formation in vivo In addition, we observed a marked increase in inflammation and IL6 production in RASSF1A-deficient tumors. Thus, RASSF1A loss profoundly affects RAS-driven lung tumorigenesis and mitogenic signaling in vivo Deregulation of inflammatory pathways due to loss of RASSF1A may be essential for RAS-mediated tumorigenesis. These results may have considerable ramifications for future targeted therapy against RAS+/RASSF1A- tumors.Significance: A transgenic mouse model shows that suppression of RASSF1A dramatically enhances Ras-driven tumorigenesis and alters Ras signaling pathway activity.Graphical Abstract: http://cancerres.aacrjournals.org/content/canres/78/10/2614/F1.large.jpg Cancer Res; 78(10); 2614-23. ©2018 AACR.


Young people's beliefs about the harmfulness of alcohol, cannabis and tobacco for mental disorders: findings from two Australian national youth surveys.
Using cross-sectional national survey data, we assessed young peoples' beliefs about the role of alcohol, tobacco and marijuana in the prevention and treatment of mental disorders as well as the predictors of these beliefs. We also compared these findings with those from a similar survey carried out in 2006. Between January and May 2011, a national computer-assisted telephone survey was conducted on a representative sample of Australian youths aged 15-25 years. A total of 3021 young people were presented with a case vignette portraying depression, depression with suicidal thoughts, psychosis, social phobia, depression with alcohol misuse or post-traumatic stress disorder in a young person. Respondents were asked about their beliefs regarding the role of using alcohol, tobacco and marijuana in preventing or dealing with the mental disorders described in the vignettes. Level of psychological distress was assessed by the Kessler 6 scale (K6). More than 75% of respondents agreed that the three substances were harmful for the young people in the vignettes, and that not using marijuana or drinking alcohol in excess is preventive. Males, young adults and more distressed respondents were less likely to endorse these beliefs. No significant changes were observed between surveys. Most young people in Australia are aware of the negative impact of substance use on mental disorders, but a few high-risk groups remain: males, young adults and those with more psychological distress. Future public health campaigns need to target these groups and focus on translating young people's substance use beliefs into behavioural change.


New data favouring that neurotrophins are of importance in arthritis.
Neurotrophins are important in inflammation. In an article in Arthritis Research & Therapy, Barthel and collaborators give new information on the existence of neurotrophin production in the synovial tissue of arthritic joints. These findings, together with other recent findings, stress that neurotrophins should be considered important factors in arthritis. This is reinforced by the facts that they are also produced by articular chondrocytes and that receptors for these are present in the synovial tissue and on chondrocytes. The importance of neurotrophins in joints should be further studied, including examinations on the efficacy of interfering with their effects in arthritis.


[Tijuana: endemic zone of Coccidioides immitis infection].
To consider a geographical zone as endemic for Coccidioidomycosis, three requisites are necessary: 1. isolation of the causal agent, Coccidioides immitis, from the environment, 2. a prevalence of infection among the population of that area of at least 5%, and 3. the demonstration of clinical cases of coccidioidomycosis in that population. There are no previous reports on the endemicity of Coccidioidomycosis in Tijuana, Baja California, mexico. We are reporting: a) the isolation of the causal agent, C. immitis from the environment, b) the results of a survey with intradermal antigens of the fungus, with a prevalence of infection among 1 128 healthy subjects of 10%, and c) the incidence of this systemic mycosis among the population of patients of the Tijuana General Hospital during a three-year period.


Bioprosthetic aortic valve durability in the era of transcatheter aortic valve implantation.
The main limitation of bioprosthetic valves is their limited durability, which exposes the patient to the risk of aortic valve reintervention. Transcatheter aortic valve implantation (TAVI) is considered a reasonable alternative to surgical aortic valve replacement (SAVR) in patients with intermediate or high surgical risk. TAVI is now rapidly expanding towards the lower risk populations. Although the results of midterm durability of the transcatheter bioprostheses are encouraging, their long-term durability remains largely unknown. The objective of this review article is to present the definition, mechanisms, incidence, outcome and management of structural valve deterioration of aortic bioprostheses with specific emphasis on TAVI. The structural valve deterioration can be categorised into three stages: stage 1: morphological abnormalities (fibrocalcific remodelling and tear) of bioprosthesis valve leaflets without hemodynamic valve deterioration; stage 2: morphological abnormalities and moderate hemodynamic deterioration (increase in gradient and/or new onset of transvalvular regurgitation); and stage 3: morphological abnormalities and severe hemodynamic deterioration. Several specifics inherent to the TAVI including valve oversizing, manipulation, delivery, positioning and deployment may cause injuries to the valve leaflets and increase leaflet mechanical stress, which may limit the long-term durability of transcatheter bioprostheses. The selection of the type of aortic valve replacement and bioprosthesis should thus take into account the ratio between the demonstrated durability of the bioprostheses versus the life expectancy of the patient. Pending the publication of robust data on long-term durability of transcatheter bioprostheses, it appears reasonable to select SAVR with a bioprosthesis model that has well-established long-term durability in patients with low surgical risk and long life expectancy.


Assignment of Centers for Disease Control group IVc-2 to the genus Ralstonia as Ralstonia paucula sp. nov.
An integrated genotypic and phenotypic analysis of 12 Centers for Disease Control (CDC) group IVc-2 strains revealed that this taxon represents a novel species belonging to the genus Ralstonia. Comparative 16S rDNA sequence analysis allocated a representative CDC group IVc-2 strain to the Ralstonia branch of the beta subclass of the Proteobacteria. DNA-DNA hybridizations did not detect significant binding levels towards any presently known Ralstonia species, including Ralstonia pickettii. Its DNA base ratio is between 65 and 67 mol%. The name Ralstonia paucula sp. nov. is proposed, with strain LMG 3244 (= CDC E6793), isolated from a human respiratory tract, as the type strain. R. paucula can be differentiated from other Ralstonia species by whole-cell protein analysis, amplified rDNA restriction analysis and a variety of classical biochemical tests. Strains have been isolated from various human clinical and environmental sources.