Measuring knowledge structure: reliability of concept mapping assessment in medical education.
To test the reliability of concept map assessment, which can be used to assess an individual's "knowledge structure," in a medical education setting. In 2004, 52 senior residents (pediatrics and internal medicine) and fourth-year medical students at the University of California-Davis School of Medicine created separate concept maps about two different subject domains (asthma and diabetes) on two separate occasions each (four total maps). Maps were rated using four different scoring systems: structural (S; counting propositions), quality (Q; rating the quality of propositions), importance/quality (I/Q; rating importance and quality of propositions), and a hybrid system (H; combining elements of S with I/Q). The authors used generalizability theory to determine reliability. Learners (universe score) contributed 40% to 44% to total score variation for the Q, I/Q, and H scoring systems, but only 10% for the S scoring system. There was a large learner-occasion-domain interaction effect (19%-23%). Subsequent analysis of each subject domain separately demonstrated a large learner-occasion interaction effect (31%-37%) and determined that administration on four to five occasions was necessary to achieve adequate reliability. Rater variation was uniformly low. The Q, I/Q, and H scoring systems demonstrated similar reliability and were all more reliable than the S system. The findings suggest that training and practice are required to perform the assessment task, and, as administered in this study, four to five testing occasions are required to achieve adequate reliability. Further research should focus on whether alterations in the concept mapping task could allow it to be administered over fewer occasions while maintaining adequate reliability.


Plant pathogens convergently evolved to counteract redundant nodes of an NLR immune receptor network.
In plants, nucleotide-binding domain and leucine-rich repeat (NLR)-containing proteins can form receptor networks to confer hypersensitive cell death and innate immunity. One class of NLRs, known as NLR required for cell death (NRCs), are central nodes in a complex network that protects against multiple pathogens and comprises up to half of the NLRome of solanaceous plants. Given the prevalence of this NLR network, we hypothesised that pathogens convergently evolved to secrete effectors that target NRC activities. To test this, we screened a library of 165 bacterial, oomycete, nematode, and aphid effectors for their capacity to suppress the cell death response triggered by the NRC-dependent disease resistance proteins Prf and Rpi-blb2. Among 5 of the identified suppressors, 1 cyst nematode protein and 1 oomycete protein suppress the activity of autoimmune mutants of NRC2 and NRC3, but not NRC4, indicating that they specifically counteract a subset of NRC proteins independently of their sensor NLR partners. Whereas the cyst nematode effector SPRYSEC15 binds the nucleotide-binding domain of NRC2 and NRC3, the oomycete effector AVRcap1b suppresses the response of these NRCs via the membrane trafficking-associated protein NbTOL9a (Target of Myb 1-like protein 9a). We conclude that plant pathogens have evolved to counteract central nodes of the NRC immune receptor network through different mechanisms. Coevolution with pathogen effectors may have driven NRC diversification into functionally redundant nodes in a massively expanded NLR network.


Switching biological functionalities of biointerfaces via dynamic covalent bonds.
The development of biointerfaces with switchable properties is of growing interest for fabricating advanced biomaterials since the adjustment of surface properties can be made on demand. Herein, we report a highly versatile approach for the preparation of a switchable biointerface through a dynamic covalent bond (DCB). The switchability can be achieved via the reversible attaching/detaching of aldehyde end-functionalized biomacromolecules onto/from an acylhydrazide anchored substrate surface. By applying the DCB protocol, three types of well-designed aldehyde end-functionalized biomacromolecules including aldehyde-poly(styrenesulfonate)-co-poly(ethylene glycol)methyl ether methacrylate (Ald-PSP, blood compatible), aldehyde-poly([2-(meth acryloyloxy)ethyl]trimethylammonium chloride) (Ald-PMT, antibacterial), and aldehyde-poly([2-(meth acryloyloxy)ethyl]trimethylammonium chloride-co-poly(ethylene glycol)methyl ether methacrylate) (Ald-PMP, combined antifouling and antibacterial) could be reversibly and alterably immobilized on the substrate surface by switching the pH conditions. As a result, we succeeded in altering the biointerface performances; excellent blood compatibility, antibacterial ability, or combined antifouling and antibacterial capabilities could be alterably achieved on the biointerface, which made the obtained material interfaces more adaptable and capable of satisfying different biofunctional requirements. Moreover, many other properties with specific biofunctions of interests can also be achieved via designing specific aldehyde-terminated molecules.


Application of the FFLUX Force Field to Molecular Crystals: A Study of Formamide.
In this work, we present the first application of the quantum chemical topology force field FFLUX to the solid state. FFLUX utilizes Gaussian process regression machine learning models trained on data from the interacting quantum atom partitioning scheme to predict atomic energies and flexible multipole moments that change with geometry. Here, the ambient (α) and high-pressure (β) polymorphs of formamide are used as test systems and optimized using FFLUX. Optimizing the structures with increasing multipolar ranks indicates that the lattice parameters of the α phase differ by less than 5% to the experimental structure when multipole moments up to the quadrupole are used. These differences are found to be in line with the dispersion-corrected density functional theory. Lattice dynamics calculations are also found to be possible using FFLUX, yielding harmonic phonon spectra comparable to dispersion-corrected DFT while enabling larger supercells to be considered than is typically possible with first-principles calculations. These promising results indicate that FFLUX can be used to accurately determine properties of molecular solids that are difficult to access using DFT, including the structural dynamics, free energies, and properties at finite temperature.


Depletion of Intracellular Glutamine Pools Triggers Toxoplasma gondii Stage Conversion in Human Glutamatergic Neurons.
Toxoplasma gondii chronically infects the brain as latent cysts containing bradyzoites and causes various effects in the host. Recently, the molecular mechanisms of cyst formation in the mouse brain have been elucidated, but those in the human brain remain largely unknown. Here, we show that abnormal glutamine metabolism caused by both interferon-γ (IFN-γ) stimulation and T. gondii infection induce cyst formation in human neuroblastoma cells regardless of the anti-T. gondii host factor nitric oxide (NO) level or Indoleamine 2,3-dioxygenase-1 (IDO1) expression. IFN-γ stimulation promoted intracellular glutamine degradation in human neuronal cells. Additionally, T. gondii infection inhibited the mRNA expression of the host glutamine transporters SLC38A1 and SLC38A2. These dual effects led to glutamine starvation and triggered T. gondii stage conversion in human neuronal cells. Furthermore, these mechanisms are conserved in human iPSC-derived glutamatergic neurons. Taken together, our data suggest that glutamine starvation in host cells is an important trigger of T. gondii stage conversion in human neurons.


Stiff skin syndrome--case report.
Stiff skin syndrome is a rare scleroderma-like disorder of unknown etiology characterized by stone-hard indurations of skin, mild hypertrichosis and limited joint mobility. No effective treatment has yet been found. Exercises and rehabilitative therapy are important in maintaining the patient's quality of life. The authors present a case of a two-year-old boy with progressive skin hardening since he was eight-month old and secondary restricted joint mobility, diagnosed as Stiff skin syndrome.


Changes in Optimism Are Associated with Changes in Health Over Time Among Older Adults.
Little is known about how optimism differs by age and changes over time, particularly among older adults. Even less is known about how changes in optimism are related to changes in physical health. We examined age differences and longitudinal changes in optimism in 9,790 older adults over a four-year period. We found an inverted U-shaped pattern between optimism and age both cross-sectionally and longitudinally, such that optimism generally increased in older adults before decreasing. Increases in optimism over a four-year period were associated with improvements in self-rated health and fewer chronic illnesses over the same time frame. The findings from the current study are consistent with changes in emotion regulation strategies employed by older adults and age-related changes in well-being.


Energy cost of exergaming: a comparison of the energy cost of 6 forms of exergaming.
To determine the relative effect of interactive digital exercise that features player movement (ie, exergames) on energy expenditure among children of various body mass indexes (BMIs; calculated as weight in kilograms divided by height in meters squared). Comparison study. GoKids Boston, a youth fitness research and training center located at University of Massachusetts, Boston. Thirty-nine boys and girls (mean [SD] age, 11.5 [2.0] years) recruited from local schools and after-school programs. Six forms of exergaming as well as treadmill walking. In addition to treadmill walking at 3 miles per hour (to convert miles to kilometers, multiply by 1.6), energy expenditure of the following exergames were examined: Dance Dance Revolution, LightSpace (Bug Invasion), Nintendo Wii (Boxing), Cybex Trazer (Goalie Wars), Sportwall, and Xavix (J-Mat). Energy expenditure was measured using the CosMed K4B2 portable metabolic cart. All forms of interactive gaming evaluated in our study increased energy expenditure above rest, with no between-group differences among normal (BMI < 85th percentile) and "at-risk" or overweight (BMI ≥ 85th percentile) children (P ≥ .05). Walking at 3 miles per hour resulted in a mean (SD) metabolic equivalent task value of 4.9 (0.7), whereas the intensity of exergaming resulted in mean (SD) metabolic equivalent task values of 4.2 (1.6) for Wii, 5.4 (1.8) for Dance Dance Revolution, 6.4 (1.6) for LightSpace, 7.0 (1.8) for Xavix, 5.9 (1.5) for Cybex Trazer, and 7.1 (1.7) for Sportwall. Enjoyment of the games was generally high but was highest for children with BMIs in the highest percentiles. All games used in our study elevated energy expenditure to moderate or vigorous intensity. Exergaming has the potential to increase physical activity and have a favorable influence on energy balance, and may be a viable alternative to traditional fitness activities for children of various BMI levels.


Aberrant RAG-mediated recombination contributes to multiple structural rearrangements in lymphoid blast crisis of chronic myeloid leukemia.
Blast crisis of chronic myeloid leukemia is associated with poor survival and the accumulation of genomic lesions. Using whole-exome and/or RNA sequencing of patients at chronic phase (CP, n = 49), myeloid blast crisis (MBC, n = 19), and lymphoid blast crisis (LBC, n = 20), we found 25 focal gene deletions and 14 fusions in 24 patients in BC. Deletions predominated in LBC (83% of structural variants). Transcriptional analysis identified the upregulation of genes involved in V(D)J recombination, including RAG1/2 and DNTT in LBC. RAG recombination is a reported mediator of IKZF1 deletion. We investigated the extent of RAG-mediated genomic lesions in BC. Molecular hallmarks of RAG activity; DNTT-mediated nucleotide insertions and a RAG-binding motif at structural variants were exclusively found in patients with high RAG expression. Structural variants in 65% of patients in LBC displayed these hallmarks compared with only 5% in MBC. RAG-mediated events included focal deletion and novel fusion of genes associated with hematologic cancer: IKZF1, RUNX1, CDKN2A/B, and RB1. Importantly, 8/8 patients with elevated DNTT at CP diagnosis progressed to LBC by 12 months, potentially enabling early prediction of LBC. This work confirms the central mutagenic role of RAG in LBC and describes potential clinical utility in CML management.


Impaired thiol/disulfide homeostasis in patients with mild acute pancreatitis.
The aim of this study was to compare the dynamic thiol/disulfide (SS) homeostasis and ischemia-modified albumin (IMA) concentration between healthy subjects and patients with mild acute pancreatitis (AP). A total of 28 patients with AP (AP group) and 35 age- and sex-matched healthy individuals (control group) were included in this study. Serum thiols/SS and IMA concentrations were measured and compared between the two groups. The mean serum native thiol (SH) and total thiol (TT) levels were significantly lower in the AP group than in the control group (224.7±80.3 μmol/L vs. 314.66±87.5 μmol/L, p<0.001 and 273.3±76.8 vs. 346.9±79 μmol/L, p<0.001, respectively). SS levels were significantly higher in the AP group than in the control group (24.2±11.1 μmol/L vs. 16.1±9.9 μmol/L, p<0.054). There were no differences in the IMA concentration and the mean IMA/albumin ratio (IMAR) between both the groups. We found that mild AP may affect serum thiol and SS levels, and cause impaired thiol/SS homeostasis.


Transactivation of RAGE mediates angiotensin-induced inflammation and atherogenesis.
Activation of the type 1 angiotensin II receptor (AT1) triggers proinflammatory signaling through pathways independent of classical Gq signaling that regulate vascular homeostasis. Here, we report that the AT1 receptor preformed a heteromeric complex with the receptor for advanced glycation endproducts (RAGE). Activation of the AT1 receptor by angiotensin II (Ang II) triggered transactivation of the cytosolic tail of RAGE and NF-κB-driven proinflammatory gene expression independently of the liberation of RAGE ligands or the ligand-binding ectodomain of RAGE. The importance of this transactivation pathway was demonstrated by our finding that adverse proinflammatory signaling events induced by AT1 receptor activation were attenuated when RAGE was deleted or transactivation of its cytosolic tail was inhibited. At the same time, classical homeostatic Gq signaling pathways were unaffected by RAGE deletion or inhibition. These data position RAGE transactivation by the AT1 receptor as a target for vasculoprotective interventions. As proof of concept, we showed that treatment with the mutant RAGE peptide S391A-RAGE362-404 was able to inhibit transactivation of RAGE and attenuate Ang II-dependent inflammation and atherogenesis. Furthermore, treatment with WT RAGE362-404 restored Ang II-dependent atherogenesis in Ager/Apoe-KO mice, without restoring ligand-mediated signaling via RAGE, suggesting that the major effector of RAGE activation was its transactivation.


A three-dimensional and temporo-spatial model to study invasiveness of cancer cells by heregulin and prostaglandin E2.
To study the temporal expression of motile structures and protease activity during colon cancer cell invasion by heregulin-beta1 (HRG) and prostaglandin E2 (PGE2), we have developed a three-dimensional spatial model system. HRG and PGE2 each induced the formation of well-organized, three-dimensional structures with empty spaces in the center and stimulated the expression of urokinase plasminogen activator (uPA) with differential localization of membrane-bound uPA at the focal adhesion points and leading edges of the motile cells. A specific cyclooxygenase-2 inhibitor blocked the formation of three-dimensional luminal glandular structures induced by HRG but did not block those induced by PGE2. A specific antagonist of uPA receptor completely blocked the formation of these luminal glandular structures induced by PGE2 and HRG. These findings suggest that HRG-mediated increased invasiveness of colon cancer cells is augmented at least in part by induction of PGE2 and uPA, and this augmentation may involve the formation of three-dimensional invasive structures via the uPA pathway. In addition, the three-dimensional model system presented here may have a wider application to screen the effects of therapeutic compounds and biomolecules on different spatial aspects of colonic biology, including cell growth, motility, invasion, survival, and apoptosis.


Thieno[2,3-b]pyridine derivatives: a new class of antiviral drugs against Mayaro virus.
Mayaro virus (MAYV) is an arthropod-borne virus and a member of the family Togaviridae, genus Alphavirus. Its infection leads to an acute illness accompanied by long-lasting arthralgia. To date, there are no antiviral drugs or vaccines against infection with MAYV and resources for the prevention or treatment of other alphaviruses are very limited. MAYV has served as a model to study the antiviral potential of several substances on alphavirus replication. In this work we evaluated the antiviral effect of seven new derivatives of thieno[2,3-b]pyridine against MAYV replication in a mammalian cell line. All derivatives were able to reduce viral production effectively at concentrations that were non-toxic for Vero cells. Molecular modeling assays predicted low toxicity risk and good oral bioavailability of the substances in humans. One of the molecules, selected for further study, demonstrated a strong anti-MAYV effect at early stages of replication, as it protected pre-treated cells and also during the late stages, affecting virus morphogenesis. This study is the first to demonstrate the antiviral effect of thienopyridine derivatives on MAYV replication in vitro, suggesting the potential application of these substances as antiviral molecules against alphaviruses. Additional in vivo research will be needed to expand the putative therapeutic applications.


Serotype-Specific IgG Antibody Waning after Pneumococcal Conjugate Primary Series Vaccinations with either the 10-Valent or the 13-Valent Vaccine.
The two currently available ten- and thirteen-valent pneumococcal conjugate vaccines (PCV10 and PCV13) both induce serotype-specific IgG anti-polysaccharide antibodies and are effective in preventing vaccine serotype induced invasive pneumococcal disease (IPD) as well as in reducing overall vaccine-serotype carriage and transmission and thereby inducing herd protection in the whole population. IgG levels decline after vaccination and could become too low to prevent carriage acquisition and/or pneumococcal disease. We compared the levels of 10-valent (PCV10) and 13-valent (PCV13) pneumococcal vaccine induced serum IgG antibodies at multiple time points after primary vaccinations. Data from two separate studies both performed in the Netherlands in infants vaccinated at 2, 3, and 4 months of age with either PCV10 or PCV13 were compared. Antibody levels were measured at 5, 8, and 11 months of age, during the interval between the primary immunization series and the 11-months booster dose. Serotype-specific IgG levels were determined by multiplex immunoassay. Although antibody kinetics showed significant variation between serotypes and between vaccines for the majority of the 10 shared serotypes, i.e., 1, 5, 7F, 9V, 14, 18C, and 23F, antibody concentrations were sufficiently high for both vaccines, immediately after the primary series and throughout the whole period until the booster dose. In contrast, for serotypes 4 and 19F in the PCV10 group and for serotypes 4 and 6B in the PCV13 group, IgG antibody concentrations already come within reach of the frequently used seroprotection level of 0.35 μg/mL immediately after the primary series at the five month time point and/or at eight months. This paper addresses the importance of revealing differences in serotype-specific and pneumococcal vaccine-dependent IgG antibody patterns during the interval between the primary series and the booster dose, an age period with a high IPD incidence. Trial registration: www.trialregister.nl NTR3069 and NTR2316.


Understanding the effects of intermittent shedding on the transmission of infectious diseases: example of salmonellosis in pigs.
A number of environmentally transmitted infectious diseases are characterized by intermittent infectiousness of infected hosts. However, it is unclear whether intermittent infectiousness must be explicitly accounted for in mathematical models for these diseases or if a simplified modelling approach is acceptable. To address this question we study the transmission of salmonellosis between penned pigs in a grower-finisher facility. The model considers indirect transmission, growth of free-living Salmonella within the environment, and environmental decontamination. The model is used to evaluate the role of intermittent fecal shedding by comparing the behaviour of the model with constant versus intermittent infectiousness. The basic reproduction number, [Formula: see text], is used to determine the long-term behaviour of the model regarding persistence or extinction of infection. The short-term behaviour of the model, relevant to swine production, is considered by examining the prevalence of infection at slaughter. Comparison of the two modelling approaches indicates that neglecting the intermittent pattern of infectiousness can result in biased estimates for [Formula: see text] and infection prevalence at slaughter. Therefore, models for salmonellosis or similar infections should explicitly account for the mechanism of intermittent infectiousness.


Insomnia Patients With Objective Short Sleep Duration Have a Blunted Response to Cognitive Behavioral Therapy for Insomnia.
This study examined whether individuals with insomnia and objective short sleep duration <6 h, a subgroup with greater risks of adverse health outcomes, differ in their response to cognitive-behavioral therapy for insomnia (CBT-I) when compared to individuals with insomnia and normal sleep duration ≥6 h. Secondary analyses of a randomized, clinical trial with 60 adult participants (n = 31 women) from a single academic medical center. Outpatient treatment lasted 8 weeks, with a final follow-up conducted at 6 months. Mixed-effects models controlling for age, sex, CBT-I treatment group assignment, and treatment provider examined sleep parameters gathered via actigraphy, sleep diaries, and an Insomnia Symptom Questionnaire (ISQ) across the treatment and follow-up period. Six months post-CBT-I treatment, individuals with insomnia and normal sleep duration ≥6 h fared significantly better on clinical improvement milestones than did those with insomnia and short sleep duration <6 h. Specifically, individuals with insomnia and normal sleep duration had significantly higher insomnia remission (ISQ < 36.5; χ2[1, N = 60] = 44.72, p < .0001), more normative sleep efficiency (SE) on actigraphy (SE > 80%; χ2[1, N = 60] = 21, p < .0001), normal levels of middle of the night wake after sleep onset (MWASO) <31 minutes (χ2[1, N = 60] = 37.85, p < .0001), and a >50% decline in MWASO (χ2[1, N = 60] = 60, p < .0001) compared to individuals with insomnia and short sleep duration. Additionally, those with insomnia and normal sleep duration had more success decreasing their total wake time (TWT) at the 6-month follow-up compared to those with insomnia and short sleep duration (χ2[2, N = 60] = 44.1, p < .0001). Receiver-operating characteristic curve analysis found that using a 6-h cutoff with actigraphy provided a 95.7% sensitivity and 91.9% specificity for determining insomnia remission, with the area under the curve = 0.986. Findings suggest that individuals with insomnia and objective short sleep duration <6 h are significantly less responsive to CBT-I than those with insomnia and normal sleep duration ≥6 h. Using an actigraphy TST cutoff of 6 hours to classify sleep duration groups was highly accurate and provided good discriminant value for determining insomnia remission.


Thoracoscopic major lung resection--indications, technique, and early results: experience from two centers in Asia.
The application of video-assisted thoracic surgery (VATS) to major lung resection is controversial. We reviewed our combined experience in this technique from two centers in Asia. From January 1993 to December 1995, 78 patients (44 male and 34 female patients with ages ranging from 16 to 85 years) successfully underwent VATS major lung resections. Selection criteria for this approach include (a) lesions < 5 cm in maximal diameter; (b) for primary lung carcinomas, clinical stage I status; (c) absence of chest wall involvement; (d) absence of pleural symphysis; and (e) complete or near complete interlobar fissures. Procedures included segmentectomy (1), lobectomy (69), bilobectomy (2), and pneumonectomy (6) together with mediastinal lymph node sampling in cases of primary malignancy. We emphasized not spreading ribs and using conventional thoracic instruments for dissection together with wound protection on specimen retrieval. There was one perioperative death and five nonfatal complications that included persistent air leak over 10 days in two patients, pneumonia in one, and persistent dysesthesia related to surgery in two. We conclude that VATS major lung resection is technically feasible. Stringent patient selection is important. Specific complications exist and special training is needed. The exact role of this approach in thoracic surgery remains to be defined by prospective randomized study compared with conventional thoracotomy with long-term follow-up.


The metabolome as a biomarker of mortality risk in the common marmoset.
Recently, the common marmoset has been proposed as a non-human primate model of aging. Their short lifespan coupled with pathologies that are similar to humans make them an ideal model to understand the genetic, metabolic, and environmental factors that influence aging and longevity. However, many of the underlying physiological changes that occur with age in the marmoset are unknown. Here, we attempt to determine if individual metabolites are predictive of future death and to recapitulate past metabolomic results after a change in environment (move across the country) was imposed on a colony of marmosets. We first determined that low levels of tryptophan metabolism metabolites were associated with risk of death in a 2-year follow-up in the animals, suggesting these metabolites may be used as future biomarkers of mortality. We also discovered that betaine metabolism and methionine metabolism are associated with aging regardless of environment for the animals, or of metabolomic assay technique. These two metabolic pathways are therefore of particular interest to examine as future targets for health and lifespan extending interventions. Many of the pathways associated with age in our first study of marmoset metabolomics were not found to have significant age effects in our second study, suggesting more work is needed to understand the reproducibility of large scale metabolomic studies in mammalian models. Overall, we were able to show that while several metabolomics markers show promise in understanding health and lifespan relationships with aging, it is possible that choice of technique for assay and reproducibility in these types of studies are still issues that need to be examined further.


Vitamin C requirement in surgical patients.
To summarize recent findings on vitamin C status and assess the requirement and optimal dose of supplementation in surgical patients. Blood vitamin C concentration falls after uncomplicated surgery and further decreases in surgical intensive care unit patients. The decline may be owing to increased demand caused by increased oxidative stress. To normalize plasma vitamin C concentration, much higher doses than the recommended daily allowance or doses recommended in parenteral nutrition guidelines are needed in these patients. In uncomplicated surgical patients, more than 500 mg/day of vitamin C may be required, with much higher doses in surgical intensive care unit patients. In uncomplicated gastrointestinal surgery, continuous parenteral administration of 500 mg/day of vitamin C reduced postoperative oxidative stress as manifested by reduced urinary excretion of isoprostane. In some studies, postoperative atrial fibrillation was prevented after cardiac surgery by perioperative vitamin C supplementation. In critically ill patients, some prospective randomized controlled trials support parenteral supplementation of high doses of vitamin C, E and trace elements. Vitamin C requirement is increased in surgical patients, and the potential advantage of supplementation is to increase the plasma and tissue levels of vitamin C and thereby reduce oxidative stress. Although some clinical benefits of high-dose vitamin C supplementation have been shown in the critically ill, the optimal dose for supplementation and the clinical benefits remain to be investigated in surgical patients.


Harmful help: the costs of backing-up behavior in teams.
Prior research on backing-up behavior has indicated that it is beneficial to teams (C. O. L. H. Porter, 2005; C. O. L. H. Porter et al., 2003). This literature has focused on how backing-up behavior aids backup recipients in tasks in which workload is unevenly distributed among team members. The authors of the present study examined different contexts of workload distribution and found that, in addition to the initial benefits to backup recipients, there are initial and subsequent costs. Backing-up behavior leads backup providers to neglect their own taskwork, especially when workload is evenly distributed. Team members who receive high amounts of backing-up behavior decrease their taskwork in a subsequent task, especially when a team member can observe their workload. These findings indicate that it is important to consider both the benefits and costs of engaging in backing-up behavior.


The role of Cdc2 feedback loop control in the DNA damage checkpoint in mammalian cells.
DNA damage inactivates cyclin-dependent kinases (CDKs) and arrests the cell cycle. Following DNA damage, the G1-S CDKs are inhibited by a mechanism involving p53-dependent induction of p21Cip1/Waf1; but how the Cdc2 is inhibited is less apparent. We found that the signal generated by the DNA damage checkpoint in G2 was dominant over that from the spindle microtubule-assembly checkpoint, because the high Cdc2 activity present in nocodazole or Taxol-arrested cells was reduced by DNA damage. Phosphorylation of the inhibitory residues in Cdc2, Thr14, and Tyr15 coincided with the inactivation of Cdc2 after DNA damage. Interpretation of this result, however, was not straightforward due to the regulation of Thr14/Tyr15 phosphorylation by feedback loops; hence, their phosphorylation can in principle result merely from the inhibition of Cdc2 activity. Consistent with this, Thr14/Tyr15 phosphorylation was induced when Cdc2 kinase activity was inhibited with butyrolactone-I. Given these complications, we undertook a more critical analysis of the mechanisms that regulate Cdc2 after DNA damage. Caffeine reversed the DNA damage-induced inhibition of Cdc2 by causing dephosphorylation of Cdc2, and this dephosphorylation still occurred even when the Cdc2 feedback loops were blocked with butyrolactone-I. These data suggest that the DNA damage checkpoint in part acts through Thr14/Tyr15 phosphorylation by a mechanism independent of Cdc2 activity, and this phosphorylation can be accentuated by the Cdc2 feedback loops involving Thr14/Tyr15 protein kinases and phosphatases. The kinase activity of the Wee1Hu Tyr15 protein kinase was unaltered after DNA damage, but the phosphatase activity of Cdc25C was reduced. Thus, the decrease in Cdc25C activity may in part account for the DNA damage-induced increase in Thr14/Tyr15 phosphorylation.


Five-factor model, technology enthusiasm and technology anxiety.
Older adults need to participate in the digital society, as societal and personal changes and what they do with the remaining time that they have in their older years has an undeniable effect on motivation, cognition and emotion. Changes in personality traits were investigated in older adults over the period 2019-2021. Technology enthusiasm and technology anxiety are attitudes that affect the relationship to the technology used. The changes in the score of technology enthusiasm and technology anxiety were the dependent variables. They were investigated with personality traits, age, gender, education, whether someone lives alone, cognitive function, digital social participation (DSP) and health literacy as predictors of the outcome. The Edwards-Nunnally index and logistic regression were used. The results indicated that DSP, lower age, lower neuroticism and higher education were indicative of less technology anxiety. High DSP and high extraversion are indicative of technology enthusiasm. DSP and attitude towards technology seem to be key in getting older adults to stay active online.


Meta-analysis of Plasmodium falciparum var Signatures Contributing to Severe Malaria in African Children and Indian Adults.
The clinical presentation of severe Plasmodium falciparum malaria differs between children and adults, but the mechanistic basis for this remains unclear. Contributing factors to disease severity include total parasite biomass and the diverse cytoadhesive properties mediated by the polymorphic var gene parasite ligand family displayed on infected erythrocytes. To explore these factors, we performed a multicohort analysis of the contribution of var expression and parasite biomass to severe malaria in two previously published pediatric cohorts in Tanzania and Malawi and an adult cohort in India. Machine learning analysis revealed independent and complementary roles for var adhesion types and parasite biomass in adult and pediatric severe malaria and showed that similar var profiles, including upregulation of group A and DC8 var, predict severe malaria in adults and children. Among adults, patients with multiorgan complications presented infections with significantly higher parasite biomass without significant differences in var adhesion types. Conversely, pediatric patients with specific complications showed distinct var signatures. Cerebral malaria patients showed broadly increased expression of var genes, in particular group A and DC8 var, while children with severe malaria anemia were classified based on high transcription of DC8 var only. This study represents the first large multisite meta-analysis of var expression, and it demonstrates the presence of common var profiles in severe malaria patients of different ages across distant geographical sites, as well as syndrome-specific disease signatures. The complex associations between parasite biomass, var adhesion type, and clinical presentation revealed here represent the most comprehensive picture so far of the relationship between cytoadhesion, parasite load, and clinical syndrome.IMPORTANCEP. falciparum malaria can cause multiple disease complications that differ by patient age. Previous studies have attempted to address the roles of parasite adhesion and biomass in disease severity; however, these studies have been limited to single geographical sites, and there is limited understanding of how parasite adhesion and biomass interact to influence disease manifestations. In this meta-analysis, we compared parasite disease determinants in African children and Indian adults. This study demonstrates that parasite biomass and specific subsets of var genes are independently associated with detrimental outcomes in both childhood and adult malaria. We also explored how parasite var adhesion types and biomass play different roles in the development of specific severe malaria pathologies, including childhood cerebral malaria and multiorgan complications in adults. This work represents the largest study to date of the role of both var adhesion types and biomass in severe malaria.


Effect of ethanol on hydrogen peroxide-induced AMPK phosphorylation.
AMP-activated protein kinase (AMPK) responds to oxidative stress. Previous work has shown that ethanol treatment of cultured hepatoma cells and of mice inhibited the activity of AMPK and reduced the amount of AMPK protein. Ethanol generates oxidative stress in the liver. Since AMPK is activated by reactive oxygen species, it seems paradoxical that ethanol would inhibit AMPK in the hepatoma cells. In an attempt to understand the mechanism whereby ethanol inhibits AMPK, we studied the effect of ethanol on AMPK activation by exogenous hydrogen peroxide. The effects of ethanol, hydrogen peroxide, and inhibitors of protein phosphatase 2A (PP2A) [either okadaic acid or PP2A small interference RNA (siRNA)] on AMPK phosphorylation and activity were examined in rat hepatoma cells (H4IIEC3) and HeLa cells. In H4IIEC3 cells, hydrogen peroxide (H(2)O(2), 1 mM) transiently increased the level of phospho-AMPK to 1.5-fold over control (P < 0.05). Similar findings were observed in HeLa cells, which do not express the upstream AMPK kinase, LKB1. H(2)O(2) markedly increased the phosphorylation of LKB1 in H4IIEC3 cells. Ethanol significantly inhibited the phosphorylation of PKC-zeta, LKB1, and AMPK caused by exposure to H(2)O(2). This inhibitory effect of ethanol required its metabolism. More importantly, the inhibitory effects of ethanol on H(2)O(2)-induced AMPK phosphorylation were attenuated by the presence of the PP2A inhibitor, okadaic acid, or PP2A siRNA. The inhibitory effect of ethanol on AMPK phosphorylation is exerted through the inhibition of PKC-zeta and LKB1 phosphorylation and the activation of PP2A.


Environmental oxygen regulates astrocyte proliferation to guide angiogenesis during retinal development.
Angiogenesis in the developing mammalian retina requires patterning cues from astrocytes. Developmental disorders of retinal vasculature, such as retinopathy of prematurity (ROP), involve arrest or mispatterning of angiogenesis. Whether these vascular pathologies involve astrocyte dysfunction remains untested. Here, we demonstrate that the major risk factor for ROP - transient neonatal exposure to excess oxygen - disrupts formation of the angiogenic astrocyte template. Exposing newborn mice to elevated oxygen (75%) suppressed astrocyte proliferation, whereas return to room air (21% oxygen) at postnatal day 4 triggered extensive proliferation, massively increasing astrocyte numbers and disturbing their spatial patterning prior to the arrival of developing vasculature. Proliferation required astrocytic HIF2α and was also stimulated by direct hypoxia (10% oxygen), suggesting that astrocyte oxygen sensing regulates the number of astrocytes produced during development. Along with astrocyte defects, return to room air also caused vascular defects reminiscent of ROP. Strikingly, these vascular phenotypes were more severe in animals that had larger numbers of excess astrocytes. Together, our findings suggest that fluctuations in environmental oxygen dysregulate molecular pathways controlling astrocyte proliferation, thereby generating excess astrocytes that interfere with retinal angiogenesis.


Role of UHRF1 in de novo DNA methylation in oocytes and maintenance methylation in preimplantation embryos.
The methylation of cytosine at CG sites in the mammalian genome is dynamically reprogrammed during gametogenesis and preimplantation development. It was previously shown that oocyte-derived DNMT1 (a maintenance methyltransferase) is essential for maintaining and propagating CG methylation at imprinting control regions in preimplantation embryos. In mammalian somatic cells, hemimethylated-CG-binding protein UHRF1 plays a critical role in maintaining CG methylation by recruiting DNMT1 to hemimethylated CG sites. However, the role of UHRF1 in oogenesis and preimplantation development is unknown. In the present study, we show that UHRF1 is mainly, but not exclusively, localized in the cytoplasm of oocytes and preimplantation embryos. However, smaller amounts of UHRF1 existed in the nucleus, consistent with the expected role in DNA methylation. We then generated oocyte-specific Uhrf1 knockout (KO) mice and found that, although oogenesis was itself unaffected, a large proportion of the embryos derived from the KO oocytes died before reaching the blastocyst stage (a maternal effect). Whole genome bisulfite sequencing revealed that blastocysts derived from KO oocytes have a greatly reduced level of CG methylation, suggesting that maternal UHRF1 is essential for maintaining CG methylation, particularly at the imprinting control regions, in preimplantation embryos. Surprisingly, UHRF1 was also found to contribute to de novo CG and non-CG methylation during oocyte growth: in Uhrf1 KO oocytes, transcriptionally-inactive regions gained less methylation, while actively transcribed regions, including the imprinting control regions, were unaffected or only slightly affected. We also found that de novo methylation was defective during the late stage of oocyte growth. To the best of our knowledge, this is the first study to demonstrate the role of UHRF1 in de novo DNA methylation in vivo. Our study reveals multiple functions of UHRF1 during the global epigenetic reprogramming of oocytes and early embryos.


The effects of oxygen content on bonding configurations and properties of low-k organosilicate glass dielectric films.
The low dielectric constant SiOC:H films of plasma enhanced chemical vapor deposition method have been developed with various precursor ratio. The reduction of the dielectric constant has been achieved by increasing the porosity in the films through the change of precursor ratio. In order to clarify the relation between dielectric constant and film porosity, the small angle X-ray scattering technique has been applied for characterizing pore size in the porous low-k dielectric films. The effects of the oxygen on the bonding configuration and electrical properties were investigated by adjusting TMS/O2 gas ratios. The porous SiOC:H film displays the small pore sizes and lower dielectric constant. It is found that the pore size of SiOC:H film is significant smaller than 1 nm and the pore size attributed to Si-O-Si cage structure change.


[Diagnostic and therapeutic utility of double balloon endoscopy: experience with 225 procedures].
Double balloon enteroscopy (DBE) is a new endoscopic method for the examination of the small intestine. To determine the diagnostic yield and therapeutic utility of DBE. All patients undergoing DBE using a Fuji-non intestinoscope for suspected small bowel diseases during a 2 1/2 year period were studied in a prospective single-center cohort study. All patients underwent prior EGD and colonoscopy. Patients underwent small bowel cleansing on the day before the procedure using a standard colon lavage solution. 225 DBE in 178 patients, (95 males, 83 females; mean age 59 years-old, range 12-93); oral route (n=160), anal (n=65). Indications (one or more per patient): GI bleeding (n=83), suspected Crohn's disease or evaluation of small bowel involvement or complications (n=35), diarrhea or malabsorption or suspected celiac disease (n=11), polyp removal in Peutz-Jeghers' syndrome or familial polyposis (n=23), tumor surveillance or search of primary tumor (n=14), abdominal pain (n=6) and miscellaneous (n=6). Mean duration of the procedure was 50 min, range 20 min to 150 min. Mean radiation exposure: 206 d Gy/cm2 (range 0-1492). The overall mean depth of small bowel insertion was 180 cm, ranging from 5 cm to the entire small bowel (650 cm). The mean depth of insertion via the oral route was 240 cm (range 20 cm to 650 cm) and via the anal route it was 65 cm (range 10 cm a 150 cm). A new diagnosis was reached in 108/178 patients (60%). Findings included: angiodysplasia, ulcerations, stenosis, polyps, portal jejunopathy, ischemic jejunitis and normal. DBE resulted in a therapeutic intervention (endoscopic, medical or surgical, excluding blood transfiusions) in 64% (115/178) of the patients. DBE was clinically useful for reaching a new diagnosis and to start new therapies, change existing therapies, and perform an operative intervention or to provide therapeutic endoscopy in two-thirds of the investigated patients. DBE is a useful and safe method to obtain tissue for diagnosis, to provide hemostasis and to perform polypectomy.


Effect of two length scales on the properties of MgB(2) for arbitrary applied magnetic field.
Experiments carried out on the intermetallic superconducting material MgB(2) have shown anomalous magnetic field dependence of upper critical field, small angle neutron scattering form factor, specific heat, critical current etc. Similarly, scanning tunnelling microscopy (STM) experiments on vortex structures have shown unusually large vortex core size and two different magnetic and spatial field scales. Also, whereas the specific heat measurements and isotope shift experiments have shown Bardeen-Cooper-Schrieffer-like (BCS-like) behaviour, the temperature dependences of the penetration depth experiments have shown non-BCS-like behaviour. These anomalous behaviours have been attributed to the multiband superconductivity of this material and the nature of the local spatial behaviour of the magnetic induction and the order parameter components having two length scales. We report an analytical investigation of the effect of two length scales on the temperature and the applied magnetic field dependence of several properties of MgB(2), such as, the penetration depth, single vortex and vortex lattice structure, vortex core radius, reversible magnetization, critical current, small angle neutron scattering form factor and the shear modulus of the vortex lattice within the framework of two-order parameter Ginzburg-Landau theory. We solve the corresponding nonlinear Ginzburg-Landau equations numerically exactly using an iterative method for arbitrary applied field H(c1) < H < H(c2), the Ginzburg-Landau parameter and vortex lattice symmetry. This enables us to compute the local spatial behaviour of the magnetic induction and the order parameters accurately for arbitrary applied field and a wide range of temperature. Comparison of the analytical results with experiments on MgB(2) gives very good agreement.


Anesthesia and the developing brain: A way forward for laboratory and clinical research.
All commonly used general anesthetics have been shown to cause neurotoxicity in animal models, including nonhuman primates. Opinion, however, remains divided over how cumulative evidence from preclinical and human studies in this field should be interpreted and its translation to current practices in pediatric anesthesia and surgery. A group of international experts in laboratory and clinical sciences recently convened in Genoa, Italy, to evaluate the current state of both laboratory and clinical research and discuss future directions for basic, translational, and clinical studies in this field. This paper describes those discussions and conclusions. A central goal identified was the importance of continuing to pursue laboratory research efforts to better understand the biological pathways underlying anesthesia neurotoxicity. The distinction between basic and translational experimental designs in this field was highlighted, and it was acknowledged that it will be important for future animal research to try to causally link structural changes with long-term cognitive abnormalities. While inherent limitations will continue to affect the ability of even large observational cohorts to determine if anesthesia impacts neurodevelopment or behavioral outcomes, the importance of conducting further large well-designed cohort studies was also emphasized. Adequately powered cohorts could clarify which populations are at increased risk, provide information on environmental and healthcare-related risk modifiers, and guide future interventional trials. If anesthetics cause structural or functional adverse neurological effects in young children, alternative or mitigating strategies need to be considered. While protective or mitigating strategies have been repeatedly studied in animals, there are currently no human data to support alternative anesthetic strategies in clinical practice. Lastly, it was noted that there is still considerable debate over the clinical relevance of anesthesia neurotoxicity, and the need to evaluate the impact of other aspects of perioperative care on neurodevelopment must also be considered.


Low-cost mobile phone microscopy with a reversed mobile phone camera lens.
The increasing capabilities and ubiquity of mobile phones and their associated digital cameras offer the possibility of extending low-cost, portable diagnostic microscopy to underserved and low-resource areas. However, mobile phone microscopes created by adding magnifying optics to the phone's camera module have been unable to make use of the full image sensor due to the specialized design of the embedded camera lens, exacerbating the tradeoff between resolution and field of view inherent to optical systems. This tradeoff is acutely felt for diagnostic applications, where the speed and cost of image-based diagnosis is related to the area of the sample that can be viewed at sufficient resolution. Here we present a simple and low-cost approach to mobile phone microscopy that uses a reversed mobile phone camera lens added to an intact mobile phone to enable high quality imaging over a significantly larger field of view than standard microscopy. We demonstrate use of the reversed lens mobile phone microscope to identify red and white blood cells in blood smears and soil-transmitted helminth eggs in stool samples.


The association between premature coronary artery disease and level of testosterone in young adult males.
Low testosterone levels in men have been associated with an increased risk of cardiovascular disease. We aimed to identify the association between serum testosterone level and premature coronary artery disease (CAD) and its predictors in young adult males. In this cross sectional study, consecutive male candidates for coronary angiography with unstable angina, no previous CAD and age ≤ 45 years were included. Serum levels of free (FT) and total testosterone (TT) as well as demographic and cardiovascular characteristics were compared between the CAD-positive and normal coronary subjects. The cutoff point for low TT was 2.5 ng/L. Additionally, the relationships between all the variables and the number of affected vessels and FT and TT and predictors of CAD were assessed. In this study, 191 patients with premature CAD were compared with 94 normal coronary subjects. Patients in the CAD group were significantly older (41.59 ± 3.79 versus 39.27 ± 4.97 years; P-value < 0.01), and had higher rates of diabetes mellitus (P-value = 0.04) and dyslipidemia (P-value = 0.01). Serum levels of FT and TT were significantly lower in the CAD group than the normal coronary subjects (P-value < 0.01 for both). The rate of subjects with low TT increased by the number of the affected vessels (p-value for trend <0.01) and there was a significant correlation between the Gensini score and FT and TT (r = -0.37, P-value < 0.01 and r = -0.34, P-value < 0.01, respectively). After adjustment for confounders, the association between low TT and CAD remained significant (Odds ratio = 4.30, 95% confidence interval: 1.99-9.32; P-value ≤ 0.001) CONCLUSION: Low levels of testosterone were associated with premature CAD and its severity in young adults.


Investigation of omeprazole and phenacetin first-pass metabolism in humans using a microscale bioreactor and pharmacokinetic models.
A new in vitro microfluidic platform (integrated insert dynamic microfluidic platform, IIDMP) allowing the co-culture of intestinal Caco-2 TC7 cells and of human primary hepatocytes was used to test the absorption and first-pass metabolism of two drugs: phenacetin and omeprazole. The metabolism of these drugs by CYP1A2, CYP2C19 and CYP3A4 was evaluated by the calculation of bioavailabilities and of intrinsic clearances using a pharmacokinetic (PK) model. To demonstrate the usefulness of the device and of the PK model, predictions were compared with in vitro and in vivo results from the literature. Based on the IIDMP experiments, hepatic in vivo clearances of phenacetin and omeprazole in the IIDMP were predicted to be 3.10 ± 0.36 and 1.46 ± 0.25 ml/min/kg body weight, respectively. This appeared lower than the in vivo observed data with values ranging between 11.9-19.6 and 5.8-7.5 ml/min/kg body weight, respectively. Then the calculated hepatic and intestinal clearances led to predicting an oral bioavailability of 0.85 and 0.77 for phenacetin and omeprazole versus 0.92 and 0.78 using separate data from the simple monoculture of Caco-2 TC7 cells and hepatocytes in Petri dishes. When compared with the in vivo data, the results of oral bioavailability were overestimated (0.37 and 0.71, respectively). The feasibility of co-culture in a device allowing the integration of intestinal absorption, intestinal metabolism and hepatic metabolism in a single model was demonstrated. Nevertheless, further experiments with other drugs are needed to extend knowledge of the device to predict oral bioavailability and intestinal first-pass metabolism.


Unravelling the Origin of the Vibronic Spectral Signatures in an Excitonically Coupled Indocarbocyanine Cy3 Dimer.
The indocarbocyanine Cy3 dye is widely used to probe the dynamics of proteins and DNA. Excitonically coupled Cy3 dimers exhibit very unique spectral signatures that depend on the interchromophoric geometrical orientation induced by the environment, making them powerful tools to infer the dynamics of their surroundings. Understanding the origin of the dimeric spectral signatures is a necessity for an accurate interpretation of the experimental results. In this work, we simulate the vibronic spectrum of an experimentally well-studied Cy3 dimer, and we explain the origin of the experimental signatures present in its linear absorption spectrum. The Franck-Condon harmonic approximations, among other tests, are used to probe the factors contributing to the spectrum. It is found that the first peak in the absorption spectrum originates from the lower energy excitonic state, while the next two peaks are vibrational progressions of the higher energy excitonic state. The polar solvent plays a crucial role in the appearance of the spectrum, being responsible for the localized S1 minimum, which results in an increased intensity of the first peak.


Impact of troop leader training on the implementation of physical activity opportunities in Girl Scout troop meetings.
Girl Scouts (GS) is a setting with large reach to target increased moderate-to-vigorous physical activity (MVPA) in girls. The Scouting Nutrition and Activity Program (SNAP) increased MVPA during intervention troop meetings; therefore, further examination of implementation of SNAP components is warranted to determine the processes contributing to intervention success. The purpose of this study is to examine variability in implementation of an active recreation (AR) policy into GS troop meetings. Troop leaders (n = 7) were randomized to receive SNAP, an intervention training on implementing a policy promoting AR, or a standard control. Meetings (seven meetings/troop) were observed, and girls in attendance (n = 76, 9-13 years, mean ± SD = 10.51 ± 1.19 years) wore ActiGraph GT1M accelerometers. Two observers attended each meeting and recorded start/stop points of AR time segments, as well as other tasks (i.e., opening/closing, snack, and Girl Scout curriculum [GSC]). Time-segmented accelerometer data were analyzed using Evenson cut points and paired with observation data. Researchers observed 181 segments (mean/day ± SD = 3.77 ± 1.24). Intervention troops implemented more AR segments/day (mean/day = 0.86; 95% CI = 0.62-1.09) than control troops (mean/day = 0.18; 95% CI = 0.00-0.39). A greater percentage of time (%time) was spent in MVPA during AR segments (mean = 10.99; 95% CI = 7.16-14.82) compared with GSC segments (mean = 0.73; 95% CI = 0.00-4.10), opening/closing (mean = 3.96; 95% CI = 0.54-7.38), and snack (mean = 0.74; 95% CI = 0.00-4.10) segments, and during opening/closing segments compared with GSC and snack segments. Intervention troops implemented more AR segments than control troops. Troop meeting patterns of PA were influenced by task. Adding time segments conducive to PA could increase the %time spent in MVPA during GS troop meetings.


Myoclonic Epilepsy with Ragged-red Fibers with Intranuclear Inclusions.
We herein report a case of myoclonic epilepsy with ragged-red fibers (MERRF) harboring a novel variant in mitochondrial cysteine transfer RNA (MT-TC). A 68-year-old woman presented with progressive myoclonic epilepsy with optic atrophy and peripheral neuropathy. A skin biopsy revealed p62-positive intranuclear inclusions. No mutations were found in the causative genes for diseases known to be related to intranuclear inclusions; however, a novel variant in MT-TC was found. The association between intranuclear inclusions and this newly identified MERRF-associated variant is unclear; however, the rare complication of intranuclear inclusions in a patient with typical MERRF symptoms should be noted for future studies.


Identical mutations in RAG1 or RAG2 genes leading to defective V(D)J recombinase activity can cause either T-B-severe combined immune deficiency or Omenn syndrome.
Omenn syndrome (OS) is an inherited disorder characterized by an absence of circulating B cells and an infiltration of the skin and the intestine by activated oligoclonal T lymphocytes, indicating that a profound defect in the lymphoid developmental program could be accountable for this condition. Inherited mutations in either the recombination activating genes RAG1 or RAG2, resulting in partial V(D)J recombinase activity, were shown to be responsible for OS. This study reports on the characterization of new RAG1/2 gene mutations in a series of 9 patients with OS. Given the occurrence of the same mutations in patients with T-B-severe combined immune deficiency or OS on 3 separate occasions, the proposal is made that an additional factor may be required in certain circumstances for the development of the Omenn phenotype. The nature of this factor is discussed.


Biological Co-treatment of H2S and reduction of CO2 to methane in an anoxic biological trickling filter upgrading biogas.
This study investigated the feasibility of co-treating H2S and CO2 in a biological trickling filter (BTF) inoculated with hydrogenotrophic methanogens (HMs) and nitrate-reducing, sulfur-oxidizing bacteria. This was accomplished by introducing a pure culture of Thiobacillus denitrificans in a BTF that was successfully upgrading a biogas mimic (60:40 CH4:CO2) to >97% methane using an enriched HM consortium. Nitrate was fed as the electron acceptor to oxidize H2S. The results revealed that a severe competition for hydrogen's electrons occurred between carbon dioxide and nitrate. Due to this competition, N:S loading rates of 16:1 were required to achieve >98% H2S removal, a ratio which is four times greater than the theoretical N:S ratio for complete sulfur oxidation. However, such high nitrate loading rates (>50 g N-NO3- m-3 h-1) had a negative impact on the BTF's biogas upgrading performance. An electron balance illustrated the increasing diversion of H2 electrons towards nitrate reduction as nitrate loading increased. Overall, this study showed that simultaneous biogas upgrading and H2S removal in a single bioreactor is possible, but that achieving high yields for both reactions requires further research in process and culture optimization.


Pharmacotherapy of relapsed metastatic testicular cancer.
Patients with metastatic testicular cancer exhibit an excellent prognosis. However, the outcome for patients with relapse after cisplatin-based chemotherapy remains unsatisfactory. Several larger studies have been recently published. To review the treatment of patients with testicular cancer after failure of first-line chemotherapy. A literature search was performed for studies investigating therapies for relapsed testicular cancer. The prognosis of patients relapsing after first-line cisplatin-based chemotherapy has improved with multimodality therapy, including conventional and high-dose chemotherapy, surgery and radiation. Prognostic factors are increasingly used to guide treatment intensity. High-dose chemotherapy has become an accepted treatment option, in particular in patients with poor risk factors at relapse. The outcome of patients with multiply relapsed or cisplatin-refractory disease remains particularly poor. Treatment of relapsed patients requires a close cooperation of medical oncologists, urologists, surgeons and radiation oncologists with extensive experience in this disease.


Spatial heterogeneity as a genetic mixing mechanism in highly philopatric colonial seabirds.
How genetic diversity is maintained in philopatric colonial systems remains unclear, and understanding the dynamic balance of philopatry and dispersal at all spatial scales is essential to the study of the evolution of coloniality. In the King penguin, Aptenodytes patagonicus, return rates of post-fledging chicks to their natal sub-colony are remarkably high. Empirical studies have shown that adults return year after year to their previous breeding territories within a radius of a few meters. Yet, little reliable data are available on intra- and inter-colonial dispersal in this species. Here, we present the first fine-scale study of the genetic structure in a king penguin colony in the Crozet Archipelago. Samples were collected from individual chicks and analysed at 8 microsatellite loci. Precise geolocation data of hatching sites and selective pressures associated with habitat features were recorded for all sampling locations. We found that despite strong natal and breeding site fidelity, king penguins retain a high degree of panmixia and genetic diversity. Yet, genetic structure appears markedly heterogeneous across the colony, with higher-than-expected inbreeding levels, and local inbreeding and relatedness hotspots that overlap predicted higher-quality nesting locations. This points towards heterogeneous population structure at the sub-colony level, in which fine-scale environmental features drive local philopatric behaviour, while lower-quality patches may act as genetic mixing mechanisms at the colony level. These findings show how a lack of global genetic structuring can emerge from small-scale heterogeneity in ecological parameters, as opposed to the classical model of homogeneous dispersal. Our results also emphasize the importance of sampling design for estimation of population parameters in colonial seabirds, as at high spatial resolution, basic genetic features are shown to be location-dependent. Finally, this study stresses the importance of understanding intra-colonial dispersal and genetic mixing mechanisms in order to better estimate species-wide gene flows and population dynamics.


Non-small cell lung cancer is characterized by dramatic changes in phospholipid profiles.
Non-small cell lung cancer (NSCLC) is the leading cause of cancer death globally. To develop better diagnostics and more effective treatments, research in the past decades has focused on identification of molecular changes in the genome, transcriptome, proteome, and more recently also the metabolome. Phospholipids, which nevertheless play a central role in cell functioning, remain poorly explored. Here, using a mass spectrometry (MS)-based phospholipidomics approach, we profiled 179 phospholipid species in malignant and matched non-malignant lung tissue of 162 NSCLC patients (73 in a discovery cohort and 89 in a validation cohort). We identified 91 phospholipid species that were differentially expressed in cancer versus non-malignant tissues. Most prominent changes included a decrease in sphingomyelins (SMs) and an increase in specific phosphatidylinositols (PIs). Also a decrease in multiple phosphatidylserines (PSs) was observed, along with an increase in several phosphatidylethanolamine (PE) and phosphatidylcholine (PC) species, particularly those with 40 or 42 carbon atoms in both fatty acyl chains together. 2D-imaging MS of the most differentially expressed phospholipids confirmed their differential abundance in cancer cells. We identified lipid markers that can discriminate tumor versus normal tissue and different NSCLC subtypes with an AUC (area under the ROC curve) of 0.999 and 0.885, respectively. In conclusion, using both shotgun and 2D-imaging lipidomics analysis, we uncovered a hitherto unrecognized alteration in phospholipid profiles in NSCLC. These changes may have important biological implications and may have significant potential for biomarker development.


[Diagnostic relevance of fine needle aspiration cytology in nodular thyroid lesions].
Fine needle aspiration cytology (FNAC) of the thyroid gland has been used as an initial investigative procedure of thyroid nodule(s) in the Department of Pathology at the Institute of Oncology of Vojvodina for more than 20 years. This procedure is rapid, inexpensive and technologically simple, yet it has found only limited, albeit increasing acceptance in medical practice in Serbia. The aim of the study was to evaluate our FNAC findings by correlating cytological results with histological diagnosis and to define the sensitivity, diagnostic accuracy and positive predictive value of FNAC. A total of 266 patients with thyroid swellings were aspirated in one year investigated period at our Institute out of whom 69 underwent surgery between May 2008 and May 2009. The cytological results correlated with clinical features, ultrasound investigations (US) and subsequent histopathological examination of the resected tissue. By the use of cytology we found out thyroid carcinoma in 10 patients, and by histopathological examination in 12. We obtained 83% sensitivity, 100% specificity and 97% of diagnostic accuracy of FNAC. The obtained results confirm the importance of FNAC in preoperative assesment of thyroid nodule.


Optical characterization of thin films by guided waves.
The study of guided waves in a thin layer allows a precise characterization of refractive index and thickness. Optical anisotropy can also be measured. We show how this technique can be applied to the characterization of a multilayer structure.


Geriatric study in Europe on health effects of air quality in nursing homes (GERIE study) profile: objectives, study protocol and descriptive data.
Indoor air pollution (IAP) constitutes a major global public health problem requiring increasing efforts in research and policymaking that may have special significance for elderly that are likely to spend most of their day indoors and appear to be particularly susceptible to adverse effects of chemical pollutants and bio-contaminants. Yet, evidence existing on the effects of IAP in elderly is scanty. The Geriatric study in Europe on health effects of air quality in nursing homes (GERIE) study aimed to assess health effects of major indoor air pollutants and thermal conditions in elderly (> 70 years) living stably in nursing homes (NH) across Europe. Respiratory effects were particularly considered as airways and lung constitute the first target of air pollutants. We describe here the rationale and the methods of the GERIE Study. 8 nursing homes were randomly selected in 7 European countries. Twenty individuals were randomly selected in each nursing home. Major indoor and outdoor air chemical pollutants (PM10, PM2.5, PM0.1, formaldehyde, NO2; O3, VOC, CO2) and bio-contaminants (moulds, allergens) were assessed objectively with standardized procedures. Major health status indicators were assessed through a standardized questionnaire, non-invasive clinical tests and blood and urine biomarkers as well as saliva for ADN. The GERIE study has given the opportunity to publish two reviews on respiratory health effects of indoor and outdoor air pollution in elderly. In addition it has provided the inventory of air quality and thermal conditions in 50 nursing homes across Europe and data on respiratory health status in 600 elderly aged 82 years in mean. Major future results will include the relationships between NH environment and health in elderly. The main long-term purpose of the GERIE study is to improve the health of elderly who permanently reside in nursing homes or of those who are exposed to indoor air pollution because of reduced mobility.


A Visual Diagnosis: Lipodystrophy.
Lipodystrophy syndromes are rare metabolic disorders characterized by local or generalized loss of adipose tissue, resulting in insulin resistance, dyslipidemia, and cosmetic disfiguration. The lipodystrophic phenotype is highly variable, with partial lipodystrophy often missed or misdiagnosed as other diseases from a lack of a proper physical examination and low physician awareness. Correct diagnosis is important for optimal treatment and follow-up strategies in these patients. The use of GLP-1 analogs has not been systematically evaluated in lipodystrophy and could be a potential precision medicine therapy. We aim to make the reader, particularly generalists or endocrinologists outside of tertiary referral centers, aware of the presentation and clinical features of partial lipodystrophy, emphasize the role of a full physical examination in diagnosis, and discuss therapeutic options, including GLP-1-based glycemic management illustrated by our clinical case.


Operative approaches to the left atrium and mitral valve apparatus.
Traditional and alternative approaches to the mitral valve apparatus are presented. The median sternotomy and vertical left atriotomy approach affords adequate exposure in most cases. However, there are occasions when an alternative approach may be warranted.


Towards molecular diagnosis and targeted therapy of lymphoid malignancies.
Gene expression profiling of cancer began as a research tool but is rapidly moving towards clinical application. The diagnostic category of diffuse large B-cell lymphoma (DLBCL) can now be viewed as an amalgam of several different diseases that have distinct gene expression profiles, oncogenic mechanisms, and clinical outcomes. Other diagnostic categories such as chronic lymphocytic leukemia (CLL) have a single gene expression signature that distinguishes them from other lymphoid malignancies. Nevertheless, elevated expression of a single gene, ZAP-70, is characteristic of a more aggressive subtype of CLL that may require novel treatment approaches. In mantle cell lymphoma (MCL), a quantitative measurement of gene expression associated with tumor proliferation is a powerful predictor of survival. Ultimately, the molecular diagnosis of these malignancies will identify molecular pathways that can be exploited for therapy. For example, one of the gene expression subgroups of DLBCL, termed activated B-cell like DLBCL, is characterized by constitutive activation of the nuclear factor-kappaB (NF-kappaB) signaling pathway, and interference with this pathway selectively kills these lymphoma cells. The full benefit of gene expression profiling can only be realized if we incorporate this technology into prospective clinical trials.


Hyperbaric oxygen therapy modulates serum OPG/RANKL in femoral head necrosis patients.
Hyperbaric oxygen therapy (HBOT) has beneficial effects on avascular necrosis of femoral head (ANFH), but its mechanism of action is still unclear. We investigated if HBOT upregulates serum osteoprotegerin (OPG) and/or inhibits osteoclast activation. 23 patients with unilateral ANFH at stage I, II and III consented to the study: the patients received standard HBOT. Serum OPG levels were obtained at the beginning of HBOT (T0), after 15 sessions (T1), 30 sessions (T2), after a 30-day break (T3), and after 60 sessions (T4). Magnetic resonance imaging (MRI) was obtained at T0 and about one year from the end of HBO treatments. Lesion size was compared between pre- and post-HBOT. 19 patients completed the study. HBOT reduced pain symptoms in all patients. HBOT significantly reduced lesion size in all stage I and II patients and in 2 of 11 stage III patients. HBOT increased serum OPG levels but receptor activator of nuclear factor kappa-B ligand (RANKL) levels did not change.


[Mathematical model of two pathways of light perception by the photoreceptor cell].
On the basis of the concept postulating polyfunctional role of rhodopsin in the photoreceptor cell excitation a mathematical model of two pathways of excitation was formulated. One pathway involves the appearance of Ca2+ ions in the cell, which block the sodium channels. The second one results in cGMP splitting which also brings about the blocking of sodium channels. A change in the concentration of acting agents and development of cell excitation under illumination was calculated. The calcium branch is shown to be rapid and direct, but of a low sensitivity. The cGMP branch is a slow one but has a high intensification coefficient. The action time of the calcium branch does not depend on illumination, while the time of cGMP branch decreases with the illumination rise and is in an inverse proportion to the square root of the flash intensity. Under repeated illumination the amplitude of calcium response decreases, and its time remains constant. In cGMP branch the time of the response to the second flash decreases. The amplitude of the second response depends on the intensity of the first flash, it can be smaller or larger than the amplitude of the first response.


Apramycin susceptibility of multidrug-resistant Gram-negative blood culture isolates in five countries in Southeast Asia.
Bloodstream infections (BSIs) are a leading cause of sepsis, which is a life-threatening condition that significantly contributes to the mortality of bacterial infections. Aminoglycoside antibiotics such as gentamicin or amikacin are essential medicines in the treatment of BSIs, but their clinical efficacy is increasingly being compromised by antimicrobial resistance. The aminoglycoside apramycin has demonstrated preclinical efficacy against aminoglycoside-resistant and multidrug-resistant (MDR) Gram-negative bacilli (GNB) and is currently in clinical development for the treatment of critical systemic infections. This study collected a panel of 470 MDR GNB isolates from healthcare facilities in Cambodia, Laos, Singapore, Thailand and Vietnam for a multicentre assessment of their antimicrobial susceptibility to apramycin in comparison with other aminoglycosides and colistin by broth microdilution assays. Apramycin and amikacin MICs ≤ 16 µg/mL were found for 462 (98.3%) and 408 (86.8%) GNB isolates, respectively. Susceptibility to gentamicin and tobramycin (MIC ≤ 4 µg/mL) was significantly lower at 122 (26.0%) and 101 (21.5%) susceptible isolates, respectively. Of note, all carbapenem and third-generation cephalosporin-resistant Enterobacterales, all Acinetobacter baumannii and all Pseudomonas aeruginosa isolates tested in this study appeared to be susceptible to apramycin. Of the 65 colistin-resistant isolates tested, four (6.2%) had an apramycin MIC > 16 µg/mL. Apramycin demonstrated best-in-class activity against a panel of GNB isolates with resistances to other aminoglycosides, carbapenems, third-generation cephalosporins and colistin, warranting continued consideration of apramycin as a drug candidate for the treatment of MDR BSIs.


Intracranial Pressure Trajectories: A Novel Approach to Informing Severe Traumatic Brain Injury Phenotypes.
Intracranial pressure in traumatic brain injury is dynamic and influenced by factors like injury patterns, treatments, and genetics. Existing studies use time invariant summary intracranial pressure measures thus potentially losing critical information about temporal trends. We identified longitudinal intracranial pressure trajectories in severe traumatic brain injury and evaluated whether they predicted outcome. We further interrogated the model to explore whether ABCC8 polymorphisms (a known cerebraledema regulator) differed across trajectory groups. Prospective observational cohort. Single-center academic medical center. Four-hundred four severe traumatic brain injury patients. None. We used group-based trajectory modeling to identify hourly intracranial pressure trajectories in days 0-5 post traumatic brain injury incorporating risk factor adjustment (age, sex, Glasgow Coma Scale 6score, craniectomy, primary hemorrhage pattern). We compared 6-month outcomes (Glasgow Outcome Scale, Disability Rating Scale, mortality) and ABCC8 tag-single-nucleotide polymorphisms associated with cerebral edema (rs2237982, rs7105832) across groups. Regression models determined whether trajectory groups predicted outcome. A six trajectory group model best fit the data, identifying cohorts differing in initial intracranial pressure, evolution, and number/proportion of spikes greater than 20 mm Hg. There were pattern differences in age, hemorrhage type, and craniectomy rates. ABCC8 polymorphisms differed across groups. GOS (p = 0.006), Disability Rating Scale (p = 0.001), mortality (p < 0.0001), and rs2237982 (p = 0.035) differed across groups. Unfavorable outcomes were surprisingly predicted by both low intracranial pressure trajectories and sustained intracranial hypertension. Intracranial pressure variability differed across groups (p < 0.001) and may reflect preserved/impaired intracranial elastance/compliance. We employed a novel approach investigating longitudinal/dynamic intracranial pressure patterns in traumatic brain injury. In a risk adjusted model, six groups were identified and predicted outcomes. If validated, trajectory modeling may be a first step toward developing a new, granular approach for intracranial pressure phenotyping in conjunction with other phenotyping tools like biomarkers and neuroimaging. This may be particularly relevant in light of changing traumatic brain injury demographics toward the elderly.


RGS2-mediated translational control mediates cancer cell dormancy and tumor relapse.
Slow-cycling/dormant cancer cells (SCCs) have pivotal roles in driving cancer relapse and drug resistance. A mechanistic explanation for cancer cell dormancy and therapeutic strategies targeting SCCs are necessary to improve patient prognosis, but are limited because of technical challenges to obtaining SCCs. Here, by applying proliferation-sensitive dyes and chemotherapeutics to non-small cell lung cancer (NSCLC) cell lines and patient-derived xenografts, we identified a distinct SCC subpopulation that resembled SCCs in patient tumors. These SCCs displayed major dormancy-like phenotypes and high survival capacity under hostile microenvironments through transcriptional upregulation of regulator of G protein signaling 2 (RGS2). Database analysis revealed RGS2 as a biomarker of retarded proliferation and poor prognosis in NSCLC. We showed that RGS2 caused prolonged translational arrest in SCCs through persistent eukaryotic initiation factor 2 (eIF2α) phosphorylation via proteasome-mediated degradation of activating transcription factor 4 (ATF4). Translational activation through RGS2 antagonism or the use of phosphodiesterase 5 inhibitors, including sildenafil (Viagra), promoted ER stress-induced apoptosis in SCCs in vitro and in vivo under stressed conditions, such as those induced by chemotherapy. Our results suggest that a low-dose chemotherapy and translation-instigating pharmacological intervention in combination is an effective strategy to prevent tumor progression in NSCLC patients after rigorous chemotherapy.


In vitro characterization of intra-generic inhibition of growth in Salmonella typhimurium.
The intra-generic inhibition of bacterial growth observed previously in vivo and in vitro with strains of Salmonella, Citrobacter and E. coli was studied in vitro using S. typhimurium strain F98. There was complete inhibition of multiplication of S. typhimurium when it was added to stationary-phase broth cultures of different Salmonella serotypes, but only partial inhibition when added to broth cultures of E. coli. The degree of inhibition between different mutants of F98 was affected by the numbers of bacteria of the inhibiting strain, but this was not the only factor, since exponential-phase bacterial cells were less inhibitory than stationary-phase cells. The inhibitory effect was produced at temperatures between 20 degrees C and 40 degrees C. The complete inhibition of growth observed between F98 mutants was abolished by ampicillin, rifampicin and streptomycin, but not by nalidixic acid. Inhibition was also prevented by separating the two cultures by a dialysis membrane. A TnphoA insertion mutant of F98 was produced which did not show inhibition in vitro but was still inhibitory in vivo. It is suggested that this complete inhibition of bacterial multiplication between organisms of the same genus, which is greater than that produced between organisms from different genera, is mediated by a cell surface protein.


Venous Thromboembolism Recurrence in Latvian Population: Single University Hospital Data.
Background and objectives: Recurrence of venous thromboembolism (VTE) after a primary event is common; however, no sufficient risk scores have been widely introduced in clinical practice. The aim of this study was to assess the risk factors for VTE recurrences, as well as the effect of treatment strategies on the recurrence rate in a single-center patient cohort. Materials and Methods: The prospective cohort study included consecutive patients in a single center from June 2014 till June 2018 presenting with acute VTE confirmed by imaging tests. All patients were followed up for at least one year or till death. Statistical analyses were conducted using IBM SPSS Statistics 23 and Stata 13. Competing risk of death was considered. Results: A total of 219 eligible patients were identified during the study period. Pulmonary embolism with or without deep vein thrombosis (DVT) was present in 95.9% (n = 210), isolated DVT was present in 4.1% (n = 9) of patients. The total number of documented recurrences was 13 (5.9%). Incidence rate was 5.6 per 100 person-years. Recurrent VTE predicted significantly higher mortality rate (hazard ratio (HR) 6.64 [95% CI 2.61-16.93]). In univariate analysis, active cancer was associated with higher recurrence rate (p = 0.036). In competing-risks regression model (with death as the competing risk), active cancer (subdistribution hazard ratio (SHR) 2.11 (95% CI 0.58-7.76)) did not retain statistical significance for VTE recurrence. Discontinuation and duration of anticoagulant treatment (≤6 or >6 months), and drug class in acute or long-term therapy (parenteral, vitamin K antagonist (VKA), direct oral anticoagulant (DOAC)) were not associated with recurrences (p > 0.05). Conclusions: Patients who experienced recurrent VTE had 6.6-fold higher mortality rate than patients with no recurrences. The presence of active cancer was not a statistically significant risk factor for recurrence when taking into account the competing risk of death. Duration and drug class of anticoagulation did not seem to impact recurrence rate.


An analog fiber optic link developed for electrical pulse signal transmission.
The electromagnetic interference (EMI) around the devices for ionizing radiation experiments is a serious concern. In order to reduce EMI and enhance the signal-to-noise ratio, a new type of analog fiber optic link (FOL) has been developed for fast pulse signal transmission under complicated electromagnetic fields. Electrical signals are converted to optical signals through an electro-optical conversion process using the amplitude modulation mechanism. After long-distance transmission, optical signals can also be restored via photoelectric conversion. Each link has a transmitter unit connected via a fiber optic cable to the receiver unit. The transmitter unit consists of a distributed feedback laser, a driver circuit made of amplifiers, and an automatic temperature control module, while the receiver unit consists of a PIN detector, a transimpedance amplifier, a main amplifier, and a third order active low-pass filter. A prototype of this link has been fabricated and tested. The test results show that the transmission bandwidth of the analog signals is DC ∼ 155 MHz, the transmission delay is a typical 21 ns, the linear dynamic range is greater than 56 dB (692 times larger), the output noise is less than 4.5 mV, and the transmission distance is up to several kilometers, which can meet the requirements of ionizing radiation experiments. The FOL demonstrates great performance in transmission for scientific research with high quality in terms of transmission bandwidth, linearity, security, and stability.


Quantitative Insights and Visualization of Antimicrobial Tolerance in Mixed-Species Biofilms.
Biofilms are a major problem in hard-to-heal wounds. Moreover, they are composed of different species and are often tolerant to antimicrobial agents. At the same time, interspecific synergy and/or competition occurs when some bacterial species clash. For this reason, the tolerance of two dual-species wound biofilm models of Pseudomonas aeruginosa and Staphylococcus aureus or Enterococcus faecium against antimicrobials and antimicrobial dressings were analyzed quantitatively and by confocal laser scanning microscopy (CLSM). The results were compared to findings with planktonic bacteria. Octenidine-dihydrochloride/phenoxyethanol and polyhexamethylene biguanide (PHMB) irrigation solutions showed a significant, albeit delayed reduction in biofilm bacteria, while the PHMB dressing was not able to induce this effect. However, the cadexomer-iodine dressing caused a sustained reduction in and killed almost all bacteria down to 102 cfu/mL within 6 days compared to the control (1010 cfu/mL). By means of CLSM in untreated human biofilm models, it became evident that P. aeruginosa dominates over E. faecium and S. aureus. Additionally, P. aeruginosa appeared as a vast layer at the bottom of the samples, while S. aureus formed grape-like clusters. In the second model, the distribution was even clearer. Only a few E. faecium were visible, in contrast to the vast layer of P. aeruginosa. It seems that the different species avoid each other and seek their respective niches. These mixed-species biofilm models showed that efficacy and tolerance to antimicrobial substances are nearly species-independent. Their frequent application appears to be important. The bacterial wound biofilm remains a challenge in treatment and requires new, combined therapy options.


Cyclooxygenase-2 and microsomal prostaglandin E synthase-1 are overexpressed in squamous cell carcinoma of the penis.
Prostaglandin E2 (PGE2) promotes malignant growth. Cyclooxygenase (COX) catalyzes the synthesis of PGH2, which is converted, in turn, by microsomal prostaglandin E synthase (mPGES-1) to PGE2. One strategy for inhibiting carcinogenesis is to prevent PGE2 production in premalignant and malignant tissues. It is important, therefore, to determine whether enzymes involved in PGE2 biosynthesis are deregulated in neoplasia. The main purpose of this study was to determine whether amounts of COX-2 or mPGES-1 were increased in intraepithelial neoplasia or squamous cell carcinoma (SCC) of the penis. Because human papillomavirus (HPV) has been linked to the development of penile SCC, a secondary objective was to determine whether COX-2 was overexpressed in SCC arising in an HPV16 transgenic mouse. Immunohistochemistry and immunoblotting were used to evaluate the expression of COX-2 and mPGES-1 in benign and malignant lesions including metastases to lymph nodes. Amounts of intratumoral PGE2 were quantified by enzyme immunoassay. Reverse transcription-PCR was used to determine the expression of each of the four known receptors (EP(1-4)) for PGE2. Immunohistochemistry demonstrated increased expression of COX-2 and mPGES-1 in dysplasia, carcinoma in situ, invasive SCC, and metastases to lymph nodes. Immunoblot analysis confirmed that COX-2 and mPGES-1 were consistently overexpressed in SCC. PGE2 and all four of the PGE2 receptor subtypes were detected in each of the tumor samples. Elevated levels of COX-2 were also detected in SCC arising in an HPV16 transgenic mouse. Increased amounts of COX-2 and mPGES-1 were detected in penile intraepithelial neoplasia and carcinoma. These findings provide the basis for evaluating whether inhibiting COX-2 will be useful in the prevention or treatment of penile SCC.


Epidemiology of Cleistanthus collinus (oduvan) poisoning: clinical features and risk factors for mortality.
Cleistanthus collinus (local name: oduvan) poisoning is a common suicidal poisoning method used in rural southern India. There are few studies on this issue and they have small sample sizes. This study examines the epidemiology of oduvan poisoning in a large sample, highlighting socio-demographic and clinical profile, complications, and risk factors for mortality. This study is a retrospective case series of 127 oduvan intentionally poisoned patients presenting at a tertiary care teaching hospital between the years 1990-1999. Descriptive statistics, cumulative case fatality rates and time to death from ingestion of poison were calculated. Cox regression adjusting for left truncation was used to investigate the effects of covariates on death. Patients' average (sd) age was 29.1 (10.9), 62% were female, 76% were married and 49% were housewives. The cumulative case fatality rate was 30%. The median time to death after oduvan ingestion was 3 days. Common signs and symptoms included hypokalaemia, vomiting, hyponatraemia, altered sensorium, bradycardia and abnormal ECG. There was a 58% risk reduction (95% CI: 29-75) in death with each 1 mmol/l increase in plasma potassium level. Patient's age was associated with an increased risk of death and the estimated hazard ratio for an increase of 10 years in age was 1.56 (95% CI: 1.18-2.07). Use of boiled oduvan extract was associated with an increased mortality (HR: 2.71, 95% CI: 1.17-6.32) compared to ingesting fresh leaves. Risk factors for oduvan poisoning mortality were hypokalaemia, older age, mode of consumption and an elevated risk for death with presence of chronic disease. When consumed as a boiled extract, oduvan is more lethal. We recommend close monitoring, correction of plasma potassium and appropriate supportive measures.


CE method for analyzing Salmonella typhimurium in water samples.
Salmonella typhimurium is commonly described as a food-borne pathogen. However, natural and drinking water are known to be important sources for the transmission of this pathogen in developing and developed countries. The standard method to determine Salmonella is laborious and many false positives are detected. To solve this, the present work was focused on the development of a capillary zone electrophoresis method coupled to ultraviolet detection for determination of Salmonella typhimurium in water (mineral and tap water). Separations were performed in less than 11 minutes using 4.5 mM Tris (hydroxymethyl)-aminomethane, 4.5 mM boric acid and 0.1 mM ethylene diamine tetraacetate (pH 8.4) with 0.1% v/v poly ethylene oxide as separation buffer. The precision of the method was evaluated in terms of repeatability obtaining a relative standard deviation of 10.5%. Using the proposed method Salmonella typhimurium could be separated from other bacteria that could be present in water such as Escherichia coli. Finally, the proposed methodology was applied to determine Salmonella typhimurium in tap and mineral water.


Suppression of IgE antibody production in SJL mice. I. Nonspecific suppressor T cells.
High titer and persistent antihapten IgE production in SJL mice can be obtained using appropriate immunization and radiation. Nonirradiated mice rapidly terminate this antihapten IgE production. Radiation was not necessary to prolong antihapten IgE production in other strains of mice. Termination can be obtained even in irradiated SJL mice by transferring normal SJL spleen cells. That the suppressor cells are T cells is shown by using thymocytes or cells treated with anti-Thy 1.2 and complement. No appreciable suppressive effect by normal spleen cells could be demonstrated on IgG1 production in SJL mice. The characteristic of low and transient IgE antibody response in SJL mice is inherited as a recessive trait controlled by a single Mendelian autosomal gene and is not linked to the H-2-gene complex. This characteristic does not depend on the infectivity of Nippostrongylus brasiliensis, the effect of anticarrier antibody, or the recognition of antigen.


Insights into the antimicrobial efficacy of Coleus aromaticus essential oil against food-borne microbes: Biochemical and molecular simulation approaches.
The study reported the antimicrobial efficacy of chemically characterized Coleus aromaticus essential oil (CEO) against food-borne bacteria, molds (Aspergillus flavus), aflatoxin B1 (AFB1) and explored its mechanism of action using biochemical and molecular simulation approaches. The chemical profile of CEO was explored by Gas chromatography-mass spectrometry (GC-MS) analysis, which revealed thymol (46.0%) as the major compound. The minimum inhibitory concentration values of CEO for bacterial species Escherichia coli, Salmonella enterica, Bacillus cereus, and Shigella flexneri was found to be 0.9 μl/ml, 0.7 μl/ml, 0.16 μl/ml, and 0.12 μl/ml respectively. The MIC value for A. flavus and AFB1 contamination was 0.6 μl/ml. The DPPH radical scavenging activity of CEO was recorded with IC50 0.32 μl/ml. Biochemical and computational approaches (docking and dynamics simulation) have been performed to explore the multi-faceted antimicrobial inhibitory effects of CEO at the molecular level, which shows the impairment in membrane functioning, leakage of cellular contents, release of 260-nm absorbing materials, antioxidative defense, carbon catabolism and vital genes (7AP3, Nor1, Omt1, and Vbs). The findings indicated that CEO could be used as natural antimicrobial agents against food-spoilage bacteria, A. flavus and AFB1 contamination to extend the shelf-life of food product and prevention of food-borne diseases.


Assessment of reliability and validity of the Turkish version of Heartland Forgiveness Scale.
The current study investigated the reliability and validity of the Turkish version of the Heartland Forgiveness Scale (HFS). The Turkish version of the HFS, the Ruminative Response Scale (RRS), and the Satisfaction with Life Scale (SWLS) were administered to 796 Turkish university students (430 women, 366 men) with a mean age of 20.6 yr. (SD = 2.1). Internal consistency reliability was calculated and Cronbach alpha coefficients ranged between .71 and .82. Criterion validity (Pearson correlation) between the HFS and other scales ranged from -.09 to .34. Additionally, a confirmatory factor analysis was conducted to assess the fit of the standard three-factor solution of the HFS. The results suggested that the Turkish version of the HFS had adequate internal consistency, criterion validity, and reflected the standard three-factor structure, indicating that it can be reliably used to measure forgiveness among a Turkish sample.


Ventricular dysfunction in early diabetic heart disease: detection, mechanisms and significance.
The detection of preclinical heart disease is a new direction in diabetes care. This comment describes the study by Vinereanu and co-workers in this issue of Clinical Science in which tissue Doppler echocardiography has been employed to demonstrate subtle systolic and diastolic dysfunction in Type II diabetic patients who had normal global systolic function and were free of coronary artery disease. The aetiology of early ventricular dysfunction in diabetes relates to complex intramyocardial and extramyocardial mechanisms. The initiating event may be due to insulin resistance, and involves abnormal myocardial substrate utilization and uncoupling of mitochondrial oxidative phosphorylation. Dysglycaemia plays an important role via the effects of oxidative stress, protein kinase C activation and advanced glycosylation end-products on inflammatory signalling, collagen metabolism and fibrosis. Extramyocardial mechanisms involve peripheral endothelial dysfunction, arterial stiffening and autonomic neuropathy. The clinical significance of the ventricular abnormalities described is unknown. Confirmation of their prognostic importance for cardiac disease in diabetes would justify routine screening for presymptomatic ventricular dysfunction, as well as clinical trials of novel agents for correcting causal mechanisms. These considerations could also have implications for patients with obesity and the metabolic syndrome.