Probing the active center of benzaldehyde lyase with substitutions and the pseudosubstrate analogue benzoylphosphonic acid methyl ester.
Benzaldehyde lyase (BAL) catalyzes the reversible cleavage of ( R)-benzoin to benzaldehyde utilizing thiamin diphosphate and Mg (2+) as cofactors. The enzyme is important for the chemoenzymatic synthesis of a wide range of compounds via its carboligation reaction mechanism. In addition to its principal functions, BAL can slowly decarboxylate aromatic amino acids such as benzoylformic acid. It is also intriguing mechanistically due to the paucity of acid-base residues at the active center that can participate in proton transfer steps thought to be necessary for these types of reactions. Here methyl benzoylphosphonate, an excellent electrostatic analogue of benzoylformic acid, is used to probe the mechanism of benzaldehyde lyase. The structure of benzaldehyde lyase in its covalent complex with methyl benzoylphosphonate was determined to 2.49 A (Protein Data Bank entry 3D7K ) and represents the first structure of this enzyme with a compound bound in the active site. No large structural reorganization was detected compared to the complex of the enzyme with thiamin diphosphate. The configuration of the predecarboxylation thiamin-bound intermediate was clarified by the structure. Both spectroscopic and X-ray structural studies are consistent with inhibition resulting from the binding of MBP to the thiamin diphosphate in the active centers. We also delineated the role of His29 (the sole potential acid-base catalyst in the active site other than the highly conserved Glu50) and Trp163 in cofactor activation and catalysis by benzaldehyde lyase.


Genetic versus census estimators of the opportunity for sexual selection in the wild.
Abstract The existence of a direct link between intensity of sexual selection and mating-system type is widely accepted. However, the quantification of sexual selection has proven problematic. Several measures of sexual selection have been proposed, including the operational sex ratio (OSR), the breeding sex ratio (BSR), and the opportunity for sexual selection (I(mates)). For a wild population of pronghorn (Antilocapra americana), we calculated OSR and BSR. We estimated I(mates) from census data on the spatial and temporal distribution of receptive females in rut and from a multigenerational genetic pedigree. OSR and BSR indicated weak sexual selection on males, but census and pedigree I(mates) suggested stronger sexual selection on males than on females. OSR and BSR correlated with census but not pedigree estimates of I(mates), and census I(mates) did not correlate with pedigree estimates. This suggests that the behavioral mating system, as deduced from the spatial and temporal distribution of females, does not predict the genetic mating system of pronghorn. The differences we observed between estimators were primarily due to female mate sampling and choice and to the sex ratio. For most species, behavioral data are not perfectly accurate and therefore will be an insufficient alternative to using multigenerational pedigrees to quantify sexual selection.


Magnetic field enhanced microextraction rate of europium(III) with 2-thenoyltrifluoroacetone and oxalate at dodecane-water interface.
A microscopic system for the observation of reactions at a liquid-liquid interface was established, to which strong magnetic fields (0-0.4 T) could be applied with permanent magnets. In situ observation for the interfacial extraction of fluorescent and paramagnetic Eu(III) ion with 2-thenoyltrifluoroacetone (Htta) in dodecane was carried out. In the presence of oxalate (ox2-), micro-aggregates of Eu(III)-tta-ox complexes were generated in the aqueous phase before its extraction. When the micro-aggregates diffused to the dodecane-water interface, Eu(tta)3 was extracted with excess Htta in the dodecane phase. The microextraction process of the aggregates was observed as random flashes of Eu(tta)3 fluorescence at the dodecane-water interface. The single flash contained about 10(-16)-10(-14) mol of Eu(III). An application of magnetic fields made the flash frequency increase, which corresponded to an enhancement of interfacial Eu(III) extraction rate. The enhancement effect was attributable to the magnetophoresis of the paramagnetic microaggregates to the dodecane-water interface.


WIPI1, BAG1, and PEX3 Autophagy-Related Genes Are Relevant Melanoma Markers.
ROS and oxidative stress may promote autophagy; on the other hand, autophagy may help reduce oxidative damages. According to the known interplay of ROS, autophagy, and melanoma onset, we hypothesized that autophagy-related genes (ARGs) may represent useful melanoma biomarkers. We therefore analyzed the gene and protein expression of 222 ARGs in human melanoma samples, from 5 independent expression databases (overall 572 patients). Gene expression was first evaluated in the GEO database. Forty-two genes showed extremely high ability to discriminate melanoma from nevi (63 samples) according to ROC (AUC ≥ 0.85) and Mann-Whitney (p < 0.0001) analyses. The 9 genes never related to melanoma before were then in silico validated in the IST online database. BAG1, CHMP2B, PEX3, and WIPI1 confirmed a strong differential gene expression, in 355 samples. A second-round validation performed on the Human Protein Atlas database showed strong differential protein expression for BAG1, PEX3, and WIPI1 in melanoma vs control samples, according to the image analysis of 80 human histological sections. WIPI1 gene expression also showed a significant prognostic value (p < 0.0001) according to 102 melanoma patients' survival data. We finally addressed in Oncomine database whether WIPI1 overexpression is melanoma-specific. Within more than 20 cancer types, the most relevant WIPI1 expression change (p = 0.00002; fold change = 3.1) was observed in melanoma. Molecular/functional relationships of the investigated molecules with melanoma and their molecular/functional network were analyzed via Chilibot software, STRING analysis, and gene ontology enrichment analysis. We conclude that WIPI1 (AUC = 0.99), BAG1 (AUC = 1), and PEX3 (AUC = 0.93) are relevant novel melanoma markers at both gene and protein levels.


Effects of Chicory on Serum Uric Acid, Renal Function, and GLUT9 Expression in Hyperuricaemic Rats with Renal Injury and In Vitro Verification with Cells.
Hyperuricaemia (HUA) is an independent risk factor for chronic kidney disease. Urate crystals are deposited in the kidney and can cause renal tubular interstitial fibrosis, leading to renal dysfunction. Chicory extract (hereafter referred to as chicory) clearly reduced serum uric acid levels in rats with HUA induced by 10% fructose. This is the first study to observe the effect of chicory on serum uric acid levels and renal function in rats with HUA and renal injury. In vivo studies using hyperuricaemic rats with renal injury induced by yeast and adenine demonstrated that chicory decreased serum uric acid level, and its effect of delaying the progression of kidney injury was better than that of benzbromarone. In vitro cell experiments showed that this effect is related to the inhibition of GLUT9 protein expression in renal tubules and that lowering blood uric acid concentrations is one of the factors that alleviates renal damage. The results of this study indicate that chicory can be used as an alternative for alleviating renal dysfunction in hyperuricaemia.


Chronic cerebral hypoperfusion induces white matter lesions and loss of oligodendroglia with DNA fragmentation in the rat.
Cerebrovascular white matter lesions represent an age-related neurodegenerative condition that appears as a hyperintense signal on magnetic resonance images. These lesions are frequently observed in aging, hypertension and cerebrovascular disease, and are responsible for cognitive decline and gait disorders in the elderly population. In humans, cerebrovascular white matter lesions are accompanied by apoptosis of oligodendroglia, and have been thought to be caused by chronic cerebral ischemia. In the present study, we tested whether chronic cerebral hypoperfusion induces white matter lesions and apoptosis of oligodendroglia in the rat. Doppler flow meter analysis revealed an immediate reduction of cerebral blood flow ranging from 30% to 40% of that before operation; this remained at 52-64% between 7 and 30 days after operation. Transferrin-immunoreactive oligodendroglia decreased in number and the myelin became degenerated in the medial corpus callosum at 7 days and thereafter. Using the TUNEL method, the number of cells showing DNA fragmentation increased three- to eightfold between 3 and 30 days post-surgery compared to sham-operated animals. Double labeling with TUNEL and immunohistochemistry for markers of either astroglia or oligodendroglia showed that DNA fragmentation occurred in both of these glia. Messenger RNA for caspase-3 increased approximately twofold versus the sham-operated rats between 1 and 30 days post-surgery. Immunohistochemistry revealed up-regulation of caspase-3 in the oligodendroglia of the white matter, and also in the astroglia and neurons of the gray matter. Molecules involved in apoptotic signaling such as TNF-alpha and Bax were also up-regulated in glial cells. These results indicate that chronic cerebral hypoperfusion induces white matter degeneration in association with DNA fragmentation in oligodendroglia.


Compact, Energy-Efficient High-Frequency Switched Capacitor Neural Stimulator With Active Charge Balancing.
Safety and energy efficiency are two major concerns for implantable neural stimulators. This paper presents a novel high-frequency, switched capacitor (HFSC) stimulation and active charge balancing scheme, which achieves high energy efficiency and well-controlled stimulation charge in the presence of large electrode impedance variations. Furthermore, the HFSC can be implemented in a compact size without any external component to simultaneously enable multichannel stimulation by deploying multiple stimulators. The theoretical analysis shows significant benefits over the constant-current and voltage-mode stimulation methods. The proposed solution was fabricated using a 0.18 μm high-voltage technology, and occupies only 0.035 mm2 for a single stimulator. The measurement result shows 50% peak energy efficiency and confirms the effectiveness of active charge balancing to prevent the electrode dissolution.


Ribozyme termination of RNA transcripts down-regulate seed fatty acid genes in transgenic soybean.
We investigated whether termination of transcripts with a self-cleaving ribozyme can enhance nuclear retention and serve as a tool to decrease specific plant gene expression. Nuclear retention was first monitored in tobacco using the beta-glucuronidase gene terminated with either the 35S CaMV 3' untranslated sequence (UTR) or a cis-acting ribozyme. Northern blot analysis of nuclear RNA and total RNA, and in situ hybridizations showed that the ribozyme-terminated transcripts were preferentially retained in the nucleus of transgenic tobacco. Ribozyme-terminated transcripts were subsequently tested as a gene down-regulation strategy in soybean. The embryo-specific Delta-12 fatty acid desaturase FAD2-1 gene was targeted because its down-regulation elevates oleic acid content of seed storage lipids. Both ribozyme-terminated antisense and standard antisense constructs were capable of gene down-regulation, producing over 57% oleic acid compared with less than 18% in wild-type seed. Ribozyme termination cassettes were also constructed to evaluate sense transcripts for single gene down-regulation and the simultaneous down-regulation of two embryo-specific genes in soybean using a single promoter. Eight independent soybean transformants were screened that harboured standard plus sense or ribozyme terminated FAD2-1 cassette. Two of the eight ribozyme terminated transformants displayed oleic acids levels in the seed storage lipids of over 75%, while none of the standard plus sense FAD2-1 lines showed elevated oleic acid phenotypes. The dual constructs targeted FAD2-1 and the FatB gene encoding a palmitoyl-thioesterase. Five transgenic soybean lines harbouring the dual constructs had oleic acid levels, greater than 85%, and saturated fatty acids levels, less than 6%. Thus, ribozyme termination of transcripts can be utilized to specifically down-regulate endogenous gene expression in soybean.


Effects of drinker self-schema on drinking- and smoking-related information processing and behaviors.
Co-occurrence of drinking and smoking is prevalent in undergraduate students. A drinker self-schema-cognition about the self as the drinker-is a common identity in undergraduates and a well-known predictor of drinking behaviors. Given that smoking commonly occurs in the context of drinking, a drinker self-schema may be a cognitive mechanism to motivate co-occurring alcohol and tobacco use (i.e., cross-substance facilitation hypothesis). This study was to determine whether the drinker self-schema influences the processing of drinking- and smoking-related information and facilitates the co-occurrence of alcohol and tobacco use in undergraduate students who drink and smoke but do not self-identify as smokers. This study was the second phase of a 2-phase study. Of the 330 who completed phase 1 (online survey), 99 completed the phase 2 study. Phase 2 was an in-person session that included a computerized information processing task to measure endorsements and response latencies for drinking- and smoking-related attributes, and a computerized Timeline Followback that was used to measure 90-day alcohol- and tobacco-use behaviors. The 5-item drinker self-schema scale, administered in phase 1, was used to measure the strength of the drinker self-schema. A higher drinker self-schema score was associated with more endorsements of positive attributes for drinking and smoking, fewer endorsements of negative attributes for smoking, faster processing of agreements with positive alcohol-use-related attributes, higher levels of drinking and smoking, and more days of co-occurring alcohol and tobacco use. Findings provide preliminary evidence to support the cross-substance facilitation hypothesis that the drinker self-schema facilitates the processing of not only drinking-related but also smoking-related stimuli and behaviors. Undergraduates who have higher drinker self-schema scores may be vulnerable to co-occurring alcohol and tobacco use.


Non-geometric echocardiographic indices of ventricular function in patients with a Fontan circulation.
Complex anatomy and limited windows complicate echocardiographic assessments of ventricular function in Fontan patients. For the Pediatric Heart Network Fontan Cross-Sectional Study, data were acquired from which mean ventricular pressure change during isovolumetric contraction (dP/dt(ic)), Tei index, and maximal systolic annular velocity (S') could be measured. The purpose of this study was to compare these nongeometric indices of ventricular function to cardiac magnetic resonance (CMR) measurements of ventricular ejection fraction (EF). Echocardiographic and CMR studies were performed prospectively using standardized protocols; measurements were completed by core laboratories. Data from both modalities were available from 137 patients. A weak but statistically significant correlation was observed between mean dP/dt(ic) and CMR-derived EF (r = 0.20, P = .022). This correlation was strengthened when preload was taken into account (r = 0.30, P = .001). Statistically significant correlations did not exist between CMR-derived EF and the Tei index or S'. Among Fontan patients, the correlation between CMR-derived EF and nongeometric echocardiographic indices of ventricular function is not strong. Of the indices evaluated, however, mean dP/dt(ic) appears to be the best.


[Cerebral Autosomal Recessive Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CARASIL)].
Cerebral small vessel disease (CSVD) is frequently observed among the elderly and is known to cause dementia and gait disturbance associated with white matter lesions, lacunar infarcts, and cerebral hemorrhage. Molecular mechanistic studies promise to provide new insights into the pathogenesis of hereditary CSVD. Cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL) is one of the hereditary CSVDs caused by a mutation in the high-temperature requirement serine peptidase A1 (HTRA1) gene. The loss of HTRA1 protease activity increases signaling via transforming growth factor (TGF)β, thereby resulting in CARASIL. Although the CARASIL has been characterized by juvenile onset alopecia and spondylosis deformans, these features are not always observed in individuals with an HTRA1 mutation. Moreover, some HTRA1 mutations cause CSVD in heterozygous states. Therefore, the clinical features of CSVD resulting from an HTRA1 mutation extend to patients with CSVD alone or to those with dominantly inherited CSVD.


Presentation of Pediatric Unintentional Injuries at Rural Hospitals in Rwanda: A Retrospective Study.
Injuries are a leading cause of mortality among children globally, with children in low- and middle-income countries more likely to die if injured compared to children in high-income countries. Timely and high-quality care are essential to reduce injury-related morbidity and mortality. This study describes patterns, management, and outcomes of children 0-15 years presenting with unintentional injuries at three district hospitals in rural Rwanda between January 1 and December 31, 2017. Using a retrospective cross-sectional study design, we assessed the demographic and clinical characteristics, care provided, and outcomes of the children using data extracted from patient medical charts. We describe the patient population using frequencies and proportions as well as median and interquartile ranges. Of the 449 injured children who sought care at the three rural district hospitals, 66.2% (n = 297) were boys. The main causes of injury were falls (n = 261, 58.1%), burns (n = 101, 22.5%), and road traffic injuries (n = 67, 14.9%). Burns were the most common injury among children aged 0-5 years while falls were the leading injury type among the 5-15 years age group. Vital signs were inconsistently completed ranging between 23.8-89.1% of vital sign items. Of the injured children, 37.0% (n = 166) received surgery at the district hospital, general practitioners performed 80.9% (n = 114) of surgeries, 87.4% (n = 145) of operated patients received no anesthesia, and 69.3% (n = 311) were admitted to the district hospital, while 2.7% (n = 12) were transferred to tertiary facilities for higher-level care. The presentation of child injuries-namely falls, burns, and road traffic accidents-is similar to what has been reported in other sub-Saharan African countries. However, more needs to be done to improve the completion and documentation of vital signs and increase availability of surgical specialists. Finally, targeted strategies to prevent burns and motorcycle-related injuries are recommended prevention interventions for this rural population.


Biobanking strategy and sample preprocessing for integrative research in monoclonal gammopathies.
Some types of monoclonal gammopathies are typified by a very limited availability of aberrant cells. Modern research use high throughput technologies and an integrated approach for detailed characterisation of abnormal cells. This strategy requires relatively high amounts of starting material which cannot be obtained from every diagnosis without causing inconvenience to the patient. The aim of this methodological paper is to reflect our long experience with laboratory work and describe the best protocols for sample collection, sorting and further preprocessing in terms of the available number of cells and intended downstream application in monoclonal gammopathies research. Potential pitfalls are also discussed. Comparison and optimisation of freezing and sorting protocols for plasma cells in monoclonal gammopathies, followed by testing of various nucleic acid isolation and amplification techniques to establish a guideline for sample processing in haemato-oncology research. We show the average numbers of aberrant cells that can be obtained from various monoclonal gammopathies (monoclonal gammopathy of undetermined significance/light chain amyloidosis/multiple myeloma (MM)/MM circulating plasma cells/ minimal residual disease MM-10 123/22 846/305 501/68 641/4000 aberrant plasma cells of 48/30/10/16/37×106 bone marrow mononuclear cells) and the expected yield of nucleic acids provided from multiple isolation kits (DNA/RNA yield from 1 to 200×103 cells was 2.14-427/0.12-123 ng). Tested kits for parallel isolation deliver outputs comparable with kits specialised for just one type of molecule. We also present our positive experience with the whole genome amplification method, which can serve as a very powerful tool to gain complex information from a very small cell population.


Risky sexual practice and associated factors among HIV positive adults attending anti-retroviral treatment clinic at Gondar University Referral Hospital, Northwest Ethiopia.
Risky sexual practice among people living with HIV/AIDS is a public health concern because of the risk of transmission of the virus to sero-discordant partner/s. There is also the risk of re-infection with new, drug resistant viral strains between sero-concordant partners. However, there is lack of information on risky sexual practices among HIV positive adults. Therefore, this study aimed to assess risky sexual practice and associated factors among adult HIV positive clients at Gondar University Referral Hospital, Northwest Ethiopia, 2015. An institution based cross sectional study was conducted at Gondar University Referral Hospital from May to June 2015. A pretested structured questionnaire was used to collect the data. Using systematic random sampling technique, a total of 513 respondents were participated in this study. The data were entered into EPI info version 3.5.3 and transferred to SPSS version 20 for analysis. Descriptive, bivariate and multivariate analyses were done. A P-value <0.05 was considered to determine the statistical significance of the association between factors (independent variables) and risky sexual practice. The Odds ratio was also used to determine the presence and the degree of association between the dependent and independent variables. A total of 513 respondents were participated in this study. The prevalence of risky sexual practices in the past three months was 38% (95% CI: 33.3%, 42.3%). Being in the age range of 18-29 years (AOR = 2.63, 95% CI: 1.55, 4.47) and 30-39 years (AOR = 2.29, 95% CI: 1.48, 3.53), being single (AOR = 6.32, 95%CI: 2.43, 16.44),being married (AOR = 6.06, 95% CI: 2.81, 13.07), having no child (AOR = 2.58, 95% CI: 1.17, 5.72), and a CD4 count of greater than 500/mm3 were factors significantly associated with risky sexual practices. A considerable number of HIV positive clients had risky sexual practices. It is strongly recommended that the Regional Health Bureau and health service providers working at Gondar University Hospital especially in the ART Clinic need to work hard on behavioral change communication.


Terahertz metamaterials with semiconductor split-ring resonators for magnetostatic tunability.
We studied a metasurface constituted as a periodic array of semiconductor split-ring resonators. The resonance frequencies of the metasurface excited by normally incident light were found to be continuously tunable in the terahertz regime through an external magnetostatic field of suitable orientation. As such metasurfaces can be assembled into 3D metamaterials, the foregoing conclusion also applies to metamaterials comprising semiconductor split-ring resonators.


Multiple causes of asphyxia in infants at high risk for sudden infant death.
A wide range of clinical findings was present in 58 near-miss sudden infant death syndrome (SIDS) infants and 6 surviving twins of SIDS siblings. Specific investigations included: studies of gastro-oesophageal reflux and aspiration (24-hour oesophageal pH recordings, barium swallow, radionuclide 'milk-scan'); polygraphic studies of breathing, reflux, and sleep state; studies of upper airways disease (lateral airways radiography and endoscopy); detection of seizure activity by electroencephalography; evaluation of thiamine status by erythrocyte transketolase activity of venous blood. Thiamine deficiency was found in 12 of 43 tested infants; 5 of the deficient infants had a familial history of SIDS. Many potential mechanisms for asphyxia were found: idiopathic central apnoea (7 infants), tracheal obstruction from minimal tracheomalacia or aberrant innominate artery (4 infants), temporal lobe or generalised seizures (6 infants), gastro-oesophageal reflux (55 infants) with intrapulmonary aspiration (11 infants). The high incidence, severity, and timing of reflux were new findings. Reflux occurred in active and indeterminate sleep, but not in quiet sleep. The depression of respiratory reflexes by active sleep stresses the vulnerability to asphyxia. Two factors suggest that near-miss episodes are related to SIDS: the similar age distribution but earlier occurrence of near-miss episodes compared with age at death of SIDS infants, and the subsequent sudden death of 2 infants whose necropsies were consistent with SIDS.


Embedding embryos for high-resolution episcopic microscopy (HREM).
Episcopic fluorescence image capturing (EFIC) and high-resolution episcopic microscopy (HREM) are related techniques that are used to generate digital volume data and create three-dimensional (3D) images. Both techniques require specimens that are embedded in an appropriate medium, and images are captured from successive sections before removal from the embedded tissue block. EFIC detects autofluorescence emitted from the embedded tissue, whereas HREM requires the tissue to be stained with a fluorescent dye such as eosin. Different procedures are therefore necessary for embedding tissue for EFIC or HREM imaging. For HREM, the choice of fixative appears to be of little consequence. If gene expression patterns are to be visualized in addition to tissue architecture, whole-mount staining for gene expression patterns using the 4-nitro blue tetrazolium chloride (NBT)/5-bromo-4-chloro-3-indolyl phosphate (BCIP) or LacZ detection system must be performed before embedding of samples. This protocol describes the procedure for embedding E11.5 mouse embryos for HREM imaging. Processing times should be adapted for the size and nature of other specimens.


Prediction of total cholesterol: high-density-lipoprotein-cholesterol ratio in young adults.
Abnormal cholesterol fraction is an essential risk factor for atherosclerosis of large cerebral arteries in young Asians. In order to reduce the medical cost and social resource for cholesterol electrophoresis, especially in undeveloped and developing Asian countries, we evaluated the validity of Nanji's GUT score for predicting TC: HDLC ratio in this population. Results showed that GUT score only predicted 71% of them. We also tested the predictive power of CUT index, and predicting rate was 81%. Therefore, Nanji's GUT score is not an ideal surrogate for cholesterol electrophoresis. We recommend CUT index to screen for high-risk subjects till a new method can satisfy the economic pattern in Asian countries.


[What are you doing when you are doing nothing? ERP components without a cognitive task].
Two experiments, an "active" and a "passive" oddball-paradigm (1. phase) with visual and auditive stimuli were performed. In both, two stimuli with a probability of 0.7/0.3 were presented. The oddball-sequence was replaced in the 2. phase without knowledge of the subjects by a sequence, in which only targets were presented. One group of the subjects has got an active task (counting the rare stimulus), the other group was told to view the stimuli passively. The rare stimulus in the oddball-sequence evoked a larger parietal P3, the auditive stimulation additionally a larger fronto-central N1 and the visual stimulation a larger central P2. The non-expected change into the 2. phase resulted in a reverse of the ERP-lateralization between 150-300 ms: Before the change of the probability the amplitudes were more negative left than right, thereafter more negative right than left. All components and effects in the active task maintained without task, but they decreased. The differences were larger with the visual stimuli. These results suggest, that the passive paradigm could be applied to patients with motoric deficits. From a theoretical point of view these results lead to certain difficulties in the interpretation of the functional importance of the P3. It can be explained better by the context-closure theory (Desmedt, 1980; Verleger, 1988) than by the context-updating theory (Donchin, 1981).


Laser direct synthesis and patterning of silver nano/microstructures on a polymer substrate.
This study presents a novel approach for the rapid fabrication of conductive nano/microscale metal structures on flexible polymer substrate (polyimide). Silver film is simultaneously synthesized and patterned on the polyimide substrate using an advanced continuous wave (CW) laser direct writing technology and a transparent, particle-free reactive silver ion ink. The location and shape of the resulting silver patterns are written by a laser beam from a digitally controlled micromirror array device. The silver patterns fabricated by this laser direct synthesis and patterning (LDSP) process exhibit the remarkably low electrical resistivity of 2.1 μΩ cm, which is compatible to the electrical resistivity of bulk silver. This novel LDSP process requires no vacuum chamber or photomasks, and the steps needed for preparation of the modified reactive silver ink are simple and straightforward. There is none of the complexity and instability associated with the synthesis of the nanoparticles that are encountered for the conventional laser direct writing technology which involves nanoparticle sintering process. This LDSP technology is an advanced method of nano/microscale selective metal patterning on flexible substrates that is fast and environmentally benign and shows potential as a feasible process for the roll-to-roll manufacturing of large area flexible electronic devices.


Acute Kidney Injury in Pediatric Acute SARS-CoV-2 Infection and Multisystem Inflammatory Syndrome in Children (MIS-C): Is There a Difference?
Objective: To evaluate the prevalence and factors associated with the risk of acute kidney injury (AKI) in pediatric patients with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and multisystem inflammatory syndrome in children (MIS-C). Study Design: We performed a retrospective chart review of 113 patients with SARS-CoV-2 infection with or without MIS-C admitted at Children's Hospital of Michigan (CHM) from March to August 2020. Patient demographic details, laboratory data, imaging studies, echocardiography reports, and treatment data were collected. Results: Of the 92 patients included in the final analysis, 22 (24%) developed AKI with 8/22 (36%) developing stage 3 AKI. The prevalence of AKI was much higher in patients with MIS-C 15/28 (54%) vs. those with acute SARS-CoV-2 infection 7/64 (11%), (p < 0.001). Overall, when compared to patients without AKI, patients with AKI were older in age (11 vs. 6.5 years, p = 0.007), African American (86 vs. 58%, p = 0.028), had MIS-C diagnosis (68 vs. 19%, p < 0.001), required ICU admission (91 vs. 20%, p < 0.001), had cardiac dysfunction (63 vs. 16%, p < 0.001), required inotropic support (59 vs. 6%, p < 0.001) and had a greater elevation in inflammatory markers. In a multivariate analysis, requirement of inotropes [Odds Ratio (OR)-22.8, p < 0.001], African American race (OR-8.8, p = 0.023) and MIS-C diagnosis (OR-5.3, p = 0.013) were the most significant predictors for AKI. All patients had recovery of kidney function, and none required kidney replacement therapy. Conclusion: Children with acute SARS-CoV-2 infection and MIS-C are at risk for AKI, with the risk being significantly greater with MIS-C. The pathogenesis of AKI in acute SARS-CoV-2 infection appears to be a combination of both renal hypo-perfusion and direct renal parenchymal damage whereas in MIS-C, the renal injury appears to be predominantly pre-renal from cardiac dysfunction and capillary leak from a hyperinflammatory state. These factors should be considered by clinicians caring for these children with a special focus on renal protective strategies to aid in recovery and prevent additional injury to this high-risk subgroup.


Non-regular inference for dynamic weighted ordinary least squares: understanding the impact of solid food intake in infancy on childhood weight.
A dynamic treatment regime (DTR) is a set of decision rules to be applied across multiple stages of treatments. The decisions are tailored to individuals, by inputting an individual's observed characteristics and outputting a treatment decision at each stage for that individual. Dynamic weighted ordinary least squares (dWOLS) is a theoretically robust and easily implementable method for estimating an optimal DTR. As many related DTR methods, the dWOLS treatment effects estimators can be non-regular when true treatment effects are zero or very small, which results in invalid Wald-type or standard bootstrap confidence intervals. Inspired by an analysis of the effect of diet in infancy on measures of weight and body size in later childhood-a setting where the exposure is distant in time and whose effect is likely to be small-we investigate the use of the $m$-out-of-$n$ bootstrap with dWOLS as method of analysis for valid inferences of optimal DTR. We provide an extensive simulation study to compare the performance of different choices of resample size $m$ in situations where the treatment effects are likely to be non-regular. We illustrate the methodology using data from the PROmotion of Breastfeeding Intervention Trial to study the effect of solid food intake in infancy on long-term health outcomes.


Molecular enhancement of heterogeneous CO2 reduction.
The electrocatalytic carbon dioxide reduction reaction (CO2RR) addresses the need for storage of renewable energy in valuable carbon-based fuels and feedstocks, yet challenges remain in the improvement of electrosynthesis pathways for highly selective hydrocarbon production. To improve catalysis further, it is of increasing interest to lever synergies between heterogeneous and homogeneous approaches. Organic molecules or metal complexes adjacent to heterogeneous active sites provide additional binding interactions that may tune the stability of intermediates, improving catalytic performance by increasing Faradaic efficiency (product selectivity), as well as decreasing overpotential. We offer a forward-looking perspective on molecularly enhanced heterogeneous catalysis for CO2RR. We discuss four categories of molecularly enhanced strategies: molecular-additive-modified heterogeneous catalysts, immobilized organometallic complex catalysts, reticular catalysts and metal-free polymer catalysts. We introduce present-day challenges in molecular strategies and describe a vision for CO2RR electrocatalysis towards multi-carbon products. These strategies provide potential avenues to address the challenges of catalyst activity, selectivity and stability in the further development of CO2RR.


General Pediatrician-Staffed Behavioral/Developmental Access Clinic Decreases Time to Evaluation of Early Childhood Developmental Disorders.
To describe and evaluate the effectiveness of a quality improvement project to decrease wait time to evaluation for children referred to Developmental Behavioral Pediatricians (DBPs). The authors created a Behavioral/Developmental Access Clinic (BDAC) staffed by a general pediatrician (GP) and pediatric psychologist. Clinicians in the BDAC provided comprehensive developmental evaluations for children in a discrete age range (<5 yr old). We describe the establishment of the BDAC along with referrals, diagnoses, and recommended follow-up for patients seen by the GP. We used 2-tailed t tests to compare the mean time with initial evaluation for patients seen in BDAC versus a DBP. Sixty-three children were seen in BDAC over 6 months. Referrals from the BDAC included: physical/occupational/speech therapy (71%), psychology (35%), audiology (25%), genetics (14%), and neurology (8%). Five patients (8%) were diagnosed with autism spectrum disorder (ASD). Compared with time to appointment with a DBP (327 d), mean time to developmental assessment was shorter for the 45 patients who accepted earlier appointments in the BDAC (159 d), and for the 18 children seen in the BDAC as new referrals (11 d), p < .001. Follow-up with a DBP was recommended for 9 (50%) of the new patient referrals evaluated in BDAC. The BDAC allowed for earlier developmental assessment of young children, with potential for earlier diagnosis and treatment of developmental disorders, including ASD. Opportunity for initial evaluation in BDAC decreased the number of patients requiring evaluation by DBPs, improving access to this subspecialty in our institution.


Physiology and Pharmacology of Ryanodine Receptor Calcium Release Channels.
Ryanodine receptor (RyR) ion channels are essential for skeletal and cardiac muscle function. Their knockout leads to perinatal death from respiratory and cardiac failure. Acquired changes or mutations in the protein cause debilitating skeletal myopathy and cardiac arrhythmia which can be deadly. Knowledge of the pharmacology of RyR channels is central to developing effective and specific treatments of these myopathies. The ion channel is a >2.2MDa homotetamer with distinct structural and functional characteristics giving rise to a myriad of regulatory sites that are potential therapeutic targets. Australian researchers have been intimately involved in the exploration of the proteins since their identification in the mid-1980s. We discuss major aspects of RyR physiology and pharmacology that have been tackled in Australian laboratories. Specific areas of interest include ultrastructural aspects and mechanisms of RyR activation in excitation-contraction (EC) coupling and related pharmacological developments, regulation of RyRs by divalent cations, by associated proteins including the FK506-binding proteins, by redox factors and phosphorylation. We consider adverse effects of anthracycline chemotherapeutic drugs on cardiac RyRs. Phenotypes associated with RyR mutations are discussed with current and developing therapeutic approaches for treating the underlying RyR dysfunction.


Long-term follow-up of mental health and satisfaction in a Swedish sample of sperm and egg donors after open-identity donation.
How is the mental health of open-identity gamete donors and their satisfaction with their contributions 14-17 years after acceptance as a donor? The Swedish Study on Gamete Donation is a longitudinal study comprising women and men who were accepted as donors at seven Swedish university clinics between 2005 and 2008. The latest (fifth) follow-up included 215 open-identity donors (response rate 87%): 123 oocyte donors and 92 sperm donors. The donors answered a questionnaire regarding their perceptions, experiences and expectations after gamete donation 14-17 years previously. The donors were satisfied with the experience of donating, and no differences were detected between sperm and oocyte donors. Oocyte donors were more than twice as likely to feel that family and friends were proud of their donation compared with sperm donors (51% versus 23%, P < 0.001). In total, six donors regretted their donation: four oocyte donors and two sperm donors. Sperm donors were more frequently satisfied with the financial compensation compared with oocyte donors (P = 0.005). No difference in the development of symptoms of anxiety or depression was detected 14-17 years post-donation. Long-term follow-up studies on donors are important for recruiting donors, and for recipients and the children who will be conceived with donated gametes. The results from the current study indicate that donors, generally, have good mental health and do not regret their decision to donate gametes. These findings are reassuring for all parties involved.


Neonatal Hemophilia: A Rare Presentation.
Hemophilia A is a X-linked hereditary condition that lead to decreased factor VIII activity, occurs mainly in males. Decreased factor VIII activity leads to increased risk of bleeding events. During neonatal period, diagnosis is made after post-partum bleeding complication or unexpected bleeding after medical procedures. Subgaleal hemorrhage during neonatal period is a rare, severe extracranial bleeding with high mortality and usually related to traumatic labor or coagulation disorders. Subgaleal hemorrhage complications result from massive bleeding. We present a neonate with unremarkable family history and uneventful pregnancy with a vaginal delivery with no instrumentation, presenting with severe subgaleal bleeding at 52 hours of life. Aggressive support measures were implemented and bleeding managed. The unexpected bleeding lead to a coagulation study and the diagnosis of severe hemophilia A. There were no known sequelae. This case shows a rare hemophilia presentation reflecting the importance of coagulation studies when faced with unexplained severe bleeding.


Combination of Nigella sativa with Glycyrrhiza glabra and Zingiber officinale augments their protective effects on doxorubicin-induced toxicity in h9c2 cells.
The use of doxorubicin (DOX) is limited by its dose-dependent cardio toxicity in which reactive Oxygen Species (ROS) play an important role in the pathological process. The aim of this study was to evaluate the protective effect of three medicinal plants, Nigella sativa (N), Glycyrrhiza glabra (G) and Zingiber officinale (Z), and their combination (NGZ), against DOX-induced apoptosis and death in H9c2 cells. The cells were incubated with different concentrations of each extract or NGZ for 4 hr which continued in the presence or absence of 5µM doxorubicin for 24 hr. Cell viability and the apoptotic rate were determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium (MTT) and propidium iodide (PI) staining assays, respectively. The level of ROS and lipid peroxidation were measured by fluorimetric methods. Treatment with doxorubicin increased ROS generation, enhanced malondialdehyde (MDA) formation, and induced apoptosis. Co-treatment of the cells with each herb extract increased viability of cells dose-dependently with a maximum protection effect of about 30%, and their potencies were N>G>Z. The combination of the threshold dose of each extract (NGZ) produced a similar effect, which was increased dose-dependently to a maximum protection of 70%. These effects were correlated with the effects of NGZ on ROS and MDA. All of the extracts have some protective effects against DOX-induced toxicity in cardiomyocytes with similar efficacies, but with different potencies. However, NGZ produced much higher protective effect via reducing oxidative stress and inhibiting of apoptotic induction processes. Further investigations are needed to determine the effects of NGZ on DOX chemotherapy.


Pediatric Hand and Upper Extremity Injuries Presenting to Emergency Departments in the United States: Epidemiology and Health Care-Associated Costs.
Background: Upper extremity injuries represent one of the most common pediatric conditions presenting to emergency departments (EDs) in the United States. We aim to describe the epidemiology, trends, and costs of pediatric patients who present to US EDs with upper extremity injuries. Methods: Using the National Emergency Department Sample, we identified all ED encounters by patients aged <18 years associated with a primary diagnosis involving the upper extremity from 2008 to 2012. Patients were divided into 4 groups by age (≤5 years, 6-9 years, 10-13 years, and 14-17 years) and a trauma subgroup. Primary outcomes were prevalence, etiology, and associated charges. Results: In total, 11.7 million ED encounters were identified, and 89.8% had a primary diagnosis involving the upper extremity. Fracture was the most common injury type (28.2%). Dislocations were common in the youngest group (17.7%) but rare in the other 3 (range = 0.8%-1.6%). There were 73.2% of trauma-related visits, most commonly due to falls (29.9%); 96.9% of trauma patients were discharged home from the ED. There were bimodal peaks of incidence in the spring and fall and a nadir in the winter. Emergency department charges of $21.2 billion were generated during the 4 years studied. While volume of visits decreased during the study, associated charges rose by 1.21%. Conclusions: Pediatric upper extremity injuries place burden on the economy of the US health care system. Types of injuries and anticipated payers vary among age groups, and while total yearly visits have decreased over the study period, the average cost of visits has risen.


The effects of hyperosmolar solutions on cerebral arterial smooth muscle.
The present study was conducted to clarify the effect of hyperosmolar solutions on the constrictor responses of cerebral arteries to vasoactive agents, in vitro. The canine basilar arteries under resting tension were slightly relaxed with both mannitol (0.5, 1 and 2%) and sucrose (1, 2, and 4%). Constrictor responses of canine basilar arteries to 40 mM K+, 10(-7) M serotonin or 10(-6) M prostaglandin F2 alpha (PGF2 alpha) were markedly suppressed by pretreatment with either mannitol or sucrose. The rate of suppression correlated well to osmolarity changes in the Kreb's solution. When the specimens were incubated in Ca++-free medium, 10(-6) M PGF2 alpha elicited small contractions. Addition of 1 mM Ca++ to the bath promptly elicited larger contractions. The large contractions in response to the influx of extracellular Ca++ were markedly suppressed by pretreatment with mannitol or sucrose, while the small contractions induced by intracellular Ca++ were not inhibited. In addition, the contractions induced by the addition of Ca++ to the specimens depolarized with 80 mM K+ in Ca++-free medium were dose-dependently inhibited with either mannitol or sucrose, while the caffeine-induced contractions in Ca++-free medium were not altered by mannitol. These results suggest that hyperosmolar solutions produce non-specific vasodilation of cerebral arteries by inhibiting the influx of external Ca++ rather than the release of intracellularly stored Ca++. This direct vasodilatory effect may account in part for the transient increase of cerebral blood flow following administration of hyperosmolar mannitol.


Impaired glucagon-stimulated glucose output in livers of acutely uremic rats.
The role of glucagon in the pathogenesis of abnormalities of glucose metabolism associated with renal failure remains undefined. We have evaluated glucagon-stimulated glucose and cyclic AMP output and amino acid uptake in isolated perfused livers of rats with experimentally-induced ARF and sham-operated controls. ARF animals exhibited azotemia, hyperglycemia, hyperinsulinemia, and hyperglucagonemia. During stimulation with physiologic (3 X 10-10M) or supraphysiologic (3 X 10-8M) glucagon concentrations, glucose output was lower in livers of ARF rats than in those of controls, whereas cyclic AMP responses were similar or exceeded those of controls. Hepatic glycogen content was lower in rats with ARF and the stores were exhausted at the end of perfusions. Additional studies in livers of fasted animals revealed no significant differences in glucose output or amino acid uptake between ARF and control livers perfused with physiologic levels of glucagon. These experiments suggest that the decreased glucagon-stimulated glucose output in isolated perfused livers in acutely uremic rats is due primarily to glycogen depletion rather than to impaired gluconeogenesis. Normal or increased cyclic AMP responses to glucagon suggests intactness of the hormone receptor-adenylate cyclase.


Surgical induced necrotizing scleritis following intraocular lens replacement.
To report a surgical-induced necrotizing scleritis, as well as its medical and surgical management. Case-report. An 88 year-old patient with a three-day severe single-left-eye ocular pain. One-time surgery involving PPV with removal of dislocated intraocular lens and secondary implantation of iris-claw Artisan® lens was performed 6 months earlier. Visual acuity of 20/100. Slit-lamp examination revealed a 5 × 2 mm non-suppurative superior scleral defect. Empirical topical antibiotic treatment with dexamethasone, as well as oral doxycycline was started. Infectious and autoimmune diseases were ruled out. Non-infectious scleritis treatment was conducted with intravenous Methylprednisolone 3 day pulses, followed by weekly tapered Prednisone and intramuscular Methotrexate. However, 1 month after the diagnosis, the defect was worsened; hence, a heterologous scleral patch graft was performed and, days after the intervention, Adalimumab was initiated. To date, 6 months later, remains with proper scleral patch, a diary low-dose Prednisone, and spacing Adalimumab treatment. Surgery-induced necrotizing scleritis is a severe condition that compromise the ocular and visual integrity. Proper diagnosis, as well as early treatment is required to achieve remission, prevent relapses, and avoid structural complications. In refractory cases, anti-TNF-α immunotherapy associated with surgical tectonic graft interventions can achieve promising results.


Transpyloric versus gastric tube feeding for preterm infants.
Enteral feeding tubes for preterm infants may be placed in the stomach (gastric tube feeding) or in the upper small bowel (transpyloric tube feeding). There are potential advantages and disadvantages to both routes. To determine the effect of feeding via the transpyloric route versus feeding via the gastric route on feeding tolerance, growth and development and adverse consequences in preterm infants who require enteral tube feeding. The standard search strategy of the Cochrane Neonatal Review Group was used. This included electronic searches of MEDLINE and EMBASE (up to March 2007) and of The Cochrane Controlled Trials Register in The Cochrane Library (Issue 1, 2007), and searches of the references in previous reviews including cross references. Randomised or quasi-randomised controlled trials comparing transpyloric with gastric tube feeding in preterm infants. Data were extracted using the standard methods of the Cochrane Neonatal Review Group, with separate evaluation of trial quality and data extraction by each author and synthesis of data using relative risk (RR) and weighted mean difference (WMD). Data from nine trials were available. No evidence of an effect on short term growth rates was found: weight: WMD -0.7 g/week (95% confidence interval (CI) -25.2, 23.8); crown heel length: WMD -0.7 mm/week (95% CI -2.4, 1.0); head circumference: WMD 0.6 mm/week (95% CI -0.9, 2.1). Longer term growth was reported in one study. There were not any statistically significant differences between the groups in the mean body weight or occipitofrontal head circumference at three months or at six months corrected age. None of the included studies provided data on neurodevelopmental outcomes. Transpyloric feeding was associated with a greater incidence of gastro-intestinal disturbance (RR 1.45, 95% CI 1.05, 2.09). There was some evidence that feeding via the transpyloric route increased mortality (RR 2.46, 95% CI 1.36, 4.46). However, the outcomes of the study that contributed most to this finding were likely to have been affected by selective allocation of the less mature and sicker infants to transpyloric feeding. No statistically significant differences in the incidence of other adverse events, including necrotising enterocolitis, intestinal perforation, and aspiration pneumonia was found. No evidence of any beneficial effect of transpyloric feeding in preterm infants was found. However, evidence of adverse effects was noted. Feeding via the transpyloric route cannot be recommended for preterm infants.


Antibiotic consumption and ventilator-associated pneumonia rates, some parallelism but some discrepancies.
Ventilator-associated pneumonia (VAP) is a common infection in intensive care units (ICUs) but its clinical definition is neither sensitive nor specific and lacks accuracy and objectivity. New defining criteria were proposed in 2013 by the National Healthcare Safety Network (NHSN) in order to more accurately conduct surveillance and track prevention progress. Although there is a consistent trend towards a decrease in VAP incidence during the last decade, significant differences in VAP rates have been reported and are persistently lower in NHSN and other American reports (0.0 to 4.4 VAP per 1,000 ventilator-days in 2012) compared to the European Centre for Disease Prevention and Control (ECDC) data (10 VAP per 1,000 ventilator-days in 2014). In the United States, VAP has been proposed as an indicator of quality of care in public reporting, and the threat of financial penalties for this diagnosis has put pressure on hospitals to minimize VAP rates that may lead to artificial lower values, independently of patient care. Although prevention bundles may contribute for encouraging reductions in VAP incidence, both pathophysiologic and epidemiologic factors preclude a zero-VAP rate. It would be expected from the trend of reduction of VAP incidence that the consumption of antibiotics would also decrease in particular in those hospitals with lowest VAP rates. However, ICU reports show a steadily use of antibiotics for nosocomial pneumonia in 15% of patients and both ECDC and NHSN data on antibiotic consumption showed no significant trend. Knowledge of bacterial epidemiology and resistance profiles for each ICU has great relevance in order to understand trends of antibiotic use. The new NHSN criteria provide a more objective and quantitative data based VAP definition, including an antibiotic administration criterion, allowing, in theory, a more comprehensive assessment and a reportable benchmark of the observed VAP and antibiotic consumption variability.


Neurodevelopment in schizophrenia: the role of the wnt pathways.
To review the role of Wnt pathways in the neurodevelopment of schizophrenia. SYSTEMATIC PUBMED SEARCH, USING AS KEYWORDS ALL THE TERMS RELATED TO THE WNT PATHWAYS AND CROSSING THEM WITH EACH OF THE FOLLOWING AREAS: normal neurodevelopment and physiology, neurodevelopmental theory of schizophrenia, schizophrenia, and antipsychotic drug action. Neurodevelopmental, behavioural, genetic, and psychopharmacological data point to the possible involvement of Wnt systems, especially the canonical pathway, in the pathophysiology of schizophrenia and in the mechanism of antipsychotic drug action. The molecules most consistently found to be associated with abnormalities or in antipsychotic drug action are Akt1, glycogen synthase kinase3beta, and beta-catenin. However, the extent to which they contribute to the pathophysiology of schizophrenia or to antipsychotic action remains to be established. The study of the involvement of Wnt pathway abnormalities in schizophrenia may help in understanding this multifaceted clinical entity; the development of Wnt-related pharmacological targets must await the collection of more data.


atlasBREX: Automated template-derived brain extraction in animal MRI.
We proposed a generic template-derived approach for (semi-) automated brain extraction in animal MRI studies and evaluated our implementation with different animal models (macaque, marmoset, rodent) and MRI protocols (T1, T2). While conventional MR-neuroimaging studies perform brain extraction as an initial step priming subsequent image-registration from subject to template, our proposed approach propagates an anatomical template to (whole-head) individual subjects in reverse order, which is challenging due to the surrounding extracranial tissue, greater differences in contrast pattern and larger areas with field inhomogeneity. As a novel approach, the herein introduced brain extraction algorithm derives whole-brain segmentation using rigid and non-rigid deformation based on unbiased anatomical atlas building with a priori estimates from study-cohort and an initial approximate brain extraction. We evaluated our proposed method in comparison to several other technical approaches including "Marker based watershed scalper", "Brain-Extraction-Tool", "3dSkullStrip", "Primatologist-Toolbox", "Rapid Automatic Tissue Segmentation" and "Robust automatic rodent brain extraction using 3D pulse-coupled neural networks" with manual skull-stripping as reference standard. ABX demonstrated best performance with accurate (≥92%) and consistent results throughout datasets and across species, age and MRI protocols. ABX was made available to the public with documentation, templates and sample material ( https://www.github.com/jlohmeier/atlasBREX ).


Prediction of survival for older hospitalized patients: the HELP survival model. Hospitalized Elderly Longitudinal Project.
To develop and validate a model estimating the survival time of hospitalized persons aged 80 years and older. A prospective cohort study with mortality follow-up using the National Death Index. Four teaching hospitals in the US. Hospitalized patients enrolled between January 1993 and November 1994 in the Hospitalized Elderly Longitudinal Project (HELP). Patients were excluded if their length of hospital stay was 48 hours or less or if admitted electively for planned surgery. A log-normal model of survival time up to 711 days was developed with the following variables: patient demographics, disease category, nursing home residence, severity of physiologic imbalance, chart documentation of weight loss, current quality of life, exercise capacity, and functional status. We assessed whether model accuracy could be improved by including symptoms of depression or history of recent fall, serum albumin, physician's subjective estimate of prognosis, and physician and patient preferences for general approach to care. A total of 1266 patients were enrolled over a 10-month period, (median age 84.9, 61% female, 68% with one or more dependency), and 505 (40%) died during an average follow-up of more than 2 years. Important prognostic factors included the Acute Physiology Score of APACHE III collected on the third hospital day, modified Glasgow coma score, major diagnosis (ICU categories together, congestive heart failure, cancer, orthopedic, and all other), age, activities of daily living, exercise capacity, chart documentation of weight loss, and global quality of life. The Somers' Dxy for a model including these factors was 0.48 (equivalent to a receiver-operator curve (ROC) area of 0.74, suggesting good discrimination). Bootstrap estimation indicated good model validation (corrected Dxy of 0.46, ROC of 0.73). A nomogram based on this log-normal model is presented to facilitate calculation of median survival time and 10th and 90th percentile of survival time. A count of geriatric syndromes or comorbidities did not add explanatory power to the model, nor did the hospital of patient recruitment, depression, or the patient preferences for general approach to care. The physician's perception of the patient's preferences and the physician's subjective estimate of the patient's prognosis improved the estimate of survival time significantly. Accurate estimation of length of life for older hospitalized persons may be calculated using a limited amount of clinical information available from the medical chart plus a brief interview with the patient or surrogate. The accuracy of this model can be improved by including measures of the physician's perception of the patient's preferences for care and the physician's subjective estimate of prognosis.


Anti-trypanosomatid activity of ceragenins.
Cationic steroid antibiotics (CSAs), or ceragenins, are amphiphilic compounds consisting of a cholic acid backbone that is attached to several cationic amines. In this study, we tested the hypothesis that CSAs possess antiparasitic activities with minimal to no effects on mammalian cells, and thus could be used as potential therapeutic agents against pathogenic trypanosomatids. To investigate this notion, we synthesized CSAs and determined their trypanocidal and leishmanicidal activities in vitro. The 3 ceragenins assayed, i.e., CSA-8, CSA-13, and CSA-54, showed several degrees of parasiticidal activity. CSA-13 was the most effective compound against Leishmania major promastigotes and Trypanosoma cruzi trypomastigotes, at LD(50) 4.9 and 9 microM, respectively. The trypanocidal activities of these ceragenins were also assessed by infectivity experiments. We found CSA-8 was more effective on T. cruzi intracellular amastigotes when the infected host cells were treated for 24 hr (LD(50), 6.7 microM). Macrophages and LLC-MK(2) (treated for 72 hr) showed relative low susceptibility to these compounds. Our results suggest that ceragenins are indeed promising chemotherapeutic agents against trypanosomatids, but they require further investigation.


Opsonization but not pretreatment of equine macrophages with hyperimmune plasma nonspecifically enhances phagocytosis and intracellular killing of Rhodococcus equi.
Evidence regarding the efficacy of equine hyperimmune plasma to prevent pneumonia in foals caused by Rhodococcus equi is limited and conflicting. Opsonization with R. equi-specific hyperimmune plasma (HIP) will significantly increase phagocytosis and decrease intracellular replication of R. equi by alveolar macrophages (AMs) compared to normal plasma (NP). Fifteen adult Quarter Horses were used to collect bronchoalveolar lavage cells. In the first experiment, AMs from 9 horses were pretreated (incubated) with either HIP, NP, or media only (control) and then infected with nonopsonized R. equi. In a second experiment, AMs from 6 horses were infected with R. equi either opsonized with HIP or opsonized with NP. For both experiments, AMs were lysed at 0 and 48 hours and the number of viable R. equi quantified by culture were compared among groups using linear mixed-effects modeling with significance set at P < .05. Opsonization with either HIP or NP increased phagocytosis by AMs (P < .0001) and decreased intracellular survival of organisms in AMs (P < .0001). Pretreating AMs with either HIP or NP without opsonizing R. equi had no effects on phagocytosis or intracellular replication. Opsonizing R. equi with either NP or HIP decreases intracellular survival of organisms in AMs, but the effect does not appear to be enhanced by using HIP. Mechanisms other than effects on AMs must explain any clinical benefits of using HIP over NP to decrease the incidence of R. equi pneumonia in foals.


Characterization of Atypical Corticospinal Tract Microstructure and Hand Impairments in Early-Onset Hemiplegic Cerebral Palsy: Preliminary Findings.
Unilateral brain injuries occurring before at or shortly after full-term can result in hemiplegic cerebral palsy (HCP). HCP affects one side of the body and can be characterized in the hand with measures of weakness and a loss of independent hand control resulting in mirror movements. Hand impairment severity is extremely heterogeneous across individuals with HCP and the neural basis for this variability is unclear. We used diffusion MRI and tractography to investigate the relationship between structural morphology of the supraspinal corticospinal tract (CST) and the severity of two typical hand impairments experienced by individuals with HCP, grasp weakness and mirror movements. Results from nine children with HCP and eight children with typical development show that there is a significant hemispheric association between CST microstructure and hand impairment severity that may be explained by atypical development and fiber distribution of motor pathways. Further analysis in the non-lesioned (dominant) hemisphere shows significant differences for CST termination in the cortex between participants with HCP and those with typical development. These findings suggest that structural disparities at the cellular level in the seemingly unaffected hemisphere after early unilateral brain injury may be the cause of heterogeneous hand impairments seen in this population.Clinical Relevance- Quantitative measurement of the variability in hand function in individuals with HCP is necessary to represent the distinct impairments experienced by each person. Further understanding of the structural neural morphology underlying distal upper extremity motor deficits after early unilateral brain injury will help lead to the development of more specific targeted interventions that increase functional outcomes.


New perspectives in the treatment of hidradenitis suppurativa: surgery and brewer's yeast-exclusion diet.
Hidradenitis suppurativa (HS), or acne inversa, is a chronic inflammatory skin disease characterized by abscesses, fistulating sinus tracts, and scarring. The exact pathogenesis of this disease remains unclear. We present our experience with HS and a possible cause of the disease and offer a surgical and clinical treatment that allows for disease resolution. We studied 12 patients who presented with axillary and perineal fistulas and underwent surgical excision (repaired by skin grafts or healing by secondary intention) or localized treatments followed by diet modification. All patients were found to have a specific immunology IgG reaction to brewer's yeast and wheat and were put on a controlled brewer's yeast-free diet for 12 months and followed monthly. The diet demonstrated immediate stabilization of their clinical symptoms, and the skin lesions regressed over the 12-month treatment period. Similarly, all the patients demonstrated an immediate recurrence of skin lesions following accidental or voluntary consumption of beer or other foods containing brewer's yeast or wheat. The patients also demonstrated a return of their quality of life and activities, including the reestablishment of sexual activity in the patients with inguinal and perigenital lesions. The results of the investigation suggested that the patients had severe reactions to brewer's yeast. Surgery followed by the elimination of the foods containing or made with the yeast resulted in a rapid stabilization of the dermatologic manifestation and a slow, but complete, regression of the skin lesions within a year. Despite the small number of patients in our study, our study could provide the basis for extensive and multicentric studies in order to better investigate this disabling and rare disease.


Why nature chose the Mn4CaO5 cluster as water-splitting catalyst in photosystem II: a new hypothesis for the mechanism of O-O bond formation.
Resolving the questions, namely, the selection of Mn by nature to build the oxygen-evolving complex (OEC) and the presence of a cubic Mn3CaO4 structure in OEC coupled with an additional dangling Mn (Mn4) via μ-O atom are not only important to uncover the secret of water oxidation in nature, but also essential to achieve a blueprint for developing advanced water-oxidation catalysts for artificial photosynthesis. Based on the important experimental results reported so far in the literature and on our own findings, we propose a new hypothesis for the water oxidation mechanism in OEC. In this new hypothesis, we propose for the first time, a complete catalytic cycle involving a charge-rearrangement-induced MnVII-dioxo species on the dangling Mn4 during the S3 → S4 transition. Moreover, the O-O bond is formed within this MnVII-dioxo site, which is totally different from that discussed in other existing proposals.


Perspective: Fabrication of integrated organ-on-a-chip via bioprinting.
Organ-on-a-chip has emerged as a powerful platform with widespread applications in biomedical engineering, such as pathology studies and drug screening. However, the fabrication of organ-on-a-chip is still a challenging task due to its complexity. For an integrated organ-on-a-chip, it may contain four key elements, i.e., a microfluidic chip, live cells/microtissues that are cultured in this chip, components for stimulus loading to mature the microtissues, and sensors for results readout. Recently, bioprinting has been used for fabricating organ-on-a-chip as it enables the printing of multiple materials, including biocompatible materials and even live cells in a programmable manner with a high spatial resolution. Besides, all four elements for organ-on-a-chip could be printed in a single continuous procedure on one printer; in other words, the fabrication process is assembly free. In this paper, we discuss the recent advances of organ-on-a-chip fabrication by bioprinting. Light is shed on the printing strategies, materials, and biocompatibility. In addition, some specific bioprinted organs-on-chips are analyzed in detail. Because the bioprinted organ-on-a-chip is still in its early stage, significant efforts are still needed. Thus, the challenges presented together with possible solutions and future trends are also discussed.


Genetic structure in neotropical birds with different tolerance to urbanization.
Gene flow in birds can be affected by urbanization depending on natural history traits and adaptability to habitat change. Contrasting results can be expected when comparing species with opposite resilience to urbanization. In this study, we assessed genetic diversity and structure for two bird species, the urban avoider white-eared ground-sparrow, Melozone leucotis, and the urban dweller house wren Troglodytes aedon. We used seven microsatellite loci and sampled five locations with differing levels of urbanization in Costa Rica. We found considerably higher genetic structure in white-eared ground-sparrows than in house wrens. Circuit theory analyses proved a higher isolation from urban resistance for the white-eared ground-sparrow than for house wrens. These results support that urbanization is a significant barrier for gene flow in urban avoiders, in contrast to urban dweller species that showed little to no impact. Differences could be attributed to a higher plasticity in habitat and nesting site preferences in the house wren, and significant dispersal limitation for the white-eared ground-sparrow. These results emphasize the need for conservation strategies towards white-eared ground-sparrows and other urban avoider species whose habitat and connectivity have been reduced by the recent urban expansion.


Circulating cell-free DNA species affect the risk of hepatocellular carcinoma in treated chronic hepatitis B patients.
Hepatocellular carcinoma (HCC) may still develop in chronic hepatitis B (CHB) patients even under effective long-term oral antiviral therapy, but its pathogenesis in the setting of long-standing inhibition of viral replication has not been completely elucidated. We investigated whether species of circulating cell-free DNA (cfDNA) may be involved in the process of hepatocarcinogenesis in treated CHB patients. Serum samples were obtained from HBeAg-negative CHB patients with (HCC cases, n = 37) or without HCC development during the first 5 years of oral antiviral therapy (controls, n = 74). HCC cases and controls were matched 1:2 for age, sex and platelets. Determination of different circulating cfDNA species (before HCC diagnosis in HCC cases) including total cfDNA quantity, levels of Alu repeat DNA and RNase P coding DNA, copies of mitochondrial DNA and levels of 5-methyl-2'-deoxycytidine as an indicator of DNA methylation was performed. HCC cases compared with controls had higher median levels of Alu247 (123 vs 69 genomic equivalent, p = .042) and RNase P coding DNA (68 vs 15 genomic equivalent, p < .001). In contrast, median cfDNA concentration, Alu115 levels, Alu247/Alu115 ratio as an index of DNA integrity and mitochondrial DNA copies did not differ significantly between HCC cases and controls. Receiver operating characteristic curve analysis showed that levels RNase P coding DNA offered good prediction of subsequent HCC development (c-statistic: 0.80, p < .001). In conclusion, serum levels of RNase P coding DNA are increased years before HCC diagnosis and could be potentially helpful in the prediction of the HCC risk in treated HBeAg-negative CHB patients.


In vitro antifungal susceptibility profiles and genotypes of 308 clinical and environmental isolates of Cryptococcus neoformans var. grubii and Cryptococcus gattii serotype B from north-western India.
Cryptococcus neoformans and Cryptococcus gattii are aetiological agents of cryptococcosis, a major opportunistic systemic mycosis of increasing global importance. This study reports the antifungal susceptibility profiles of clinical and environmental isolates of C. neoformans var. grubii, genotype VNI/AFLP1 (n = 246), and C. gattii serotype B, genotype VGI/AFLP4 (n = 62), originating from patients and environmental sources in north-western India. All of the C. neoformans var. grubii and C. gattii isolates were mating type α. Using the broth microdilution method, both species were found to be susceptible to the antifungals tested except for two clinical C. neoformans var. grubii isolates that were resistant to 5-flucytosine (MIC >64 µg ml⁻¹). Data on the geometric mean of MICs revealed that C. gattii was significantly less susceptible than C. neoformans var. grubii to fluconazole, itraconazole and voriconazole (P<0.0001). In addition, the MIC₉₀ of C. gattii was twofold higher than that of C. neoformans var. grubii for fluconazole, itraconazole and voriconazole. However, no statistically significant difference in susceptibility of the two Cryptococcus species was observed against amphotericin B and 5-flucytosine. Furthermore, the environmental C. neoformans var. grubii isolates were significantly less susceptible to fluconazole, itraconazole and 5-flucytosine (P<0.0001) than the clinical isolates. A continued surveillance of antifungal susceptibility of clinical and environmental strains of C. neoformans and C. gattii is desirable to monitor the emergence of any resistant strains in order to ensure more successful therapy of cryptococcosis.


Breaking optical diffraction limitation using optical Hybrid-Super-Hyperlens with radially polarized light.
We propose and analyze an innovative device called "Hybrid-Super-Hyperlens". This lens is made of two hyperbolic metamaterials with different signs in their dielectric tensor and different isofrequency dispersion curves. The ability of the proposed lens to break the optical diffraction limit is demonstrated using numerical simulations (with the resolution power of about λ/6). Both a pair of nano-slits and a nano-ring can be imaged and resolved by the proposed lens using the radially polarized light source. Such a lens has great potential applications in photolithography and real-time nanoscale imaging.


A papillomavirus DNA from a cervical carcinoma and its prevalence in cancer biopsy samples from different geographic regions.
DNA from one biopsy sample of invasive cancer of the cervix contained sequences hybridizing with human papillomavirus (HPV) type 11 DNA only under nonstringent conditions. This DNA was molecularly cloned in lambda phage. Under stringent conditions of hybridization it cross-hybridized to a minor extent (less than 0.1%) with HPV types 10, 14, and 15 and showed no homology with DNA of other human HPV types. We therefore propose to designate it tentatively as HPV 16. HPV 16 DNA was used as a probe to test additional cancer biopsy samples from cervical, vulval, and penile cancer, as well as benign genital warts (condylomata acuminata) and cervical dysplasias for the presence of homologous sequences. In 61.1% (11/18) of cervical cancer samples from German patients sequences were found hybridizing with HPV 16 DNA under conditions of high stringency. In contrast, only 34.8% (8/23) of cancer biopsy samples from Kenya and Brazil revealed this DNA. Vulval and penile cancer biopsy samples hybridized to 28.6% (2/7) or 25% (1/4), respectively. Only 2 out of 33 condylomata acuminata contained HPV 16 DNA. Both positive tumors harbored in addition HPV 6 or HPV 11 DNA. The data thus indicate that HPV 16 DNA prevails in malignant tumors, rendering an accidental contamination with papillomavirus DNA from adjacent papillomas rather unlikely. The rare presence in benign genital papillomas in addition to common genital papillomaviruses suggests a dependence of HPV 16 replication on helper virus.


Crossover between Athermal Jamming and the Thermal Glass Transition of Suspensions.
The non-Newtonian flow behavior of thermal and athermal disordered systems of dispersed uniform particles at high densities have strikingly similar features. By investigating the flow curves of yield-stress fluids and colloidal glasses having different volume fractions, particle sizes, and interactions, we show that both thermal and athermal systems exhibit power-law scaling with respect to the glass and jamming point, respectively, with the same exponents. All yield-stress flow curves can be scaled onto a single universal curve using the Laplace pressure as the stress scale for athermal systems and the osmotic pressure for the thermal systems. Strikingly, the details of interparticle interactions do not matter for the rescaling, showing that they are akin to usual phase transitions of the same universality class. The rescaling allows us to predict the flow properties of these systems from the volume fraction and known material properties.


Degenerative lesions of a coronary chemoreceptor and nearby neural elements in the hearts of victims of sudden death.
A coronary chemoreceptor and its neighboring nerves and ganglia were studied by serial section from the hearts of 11 victims of sudden unexpected death. Either focal or extensive inflammatory destruction was present in the chemoreceptors of 10 of the 11 hearts, and similar abnormalities of local nerves and ganglia were present in all 11 hearts. Because an identical coronary chemoreceptor in the dog has previously been shown to be the site of origin of a powerful reflex with major influence on the electrical activity of the heart, the functional significance of the neural abnormalities found in these 11 human hearts may include important distortion of cardiac rhythm, conduction or repolarization. Future studies are needed to determine the prevalence of such lesions in the hearts of other victims of sudden death and among control subjects, as well as to determine the etiology of this special neuropathology of the heart.


Effectiveness of Rosmarinus officinalis essential oil as antihypotensive agent in primary hypotensive patients and its influence on health-related quality of life.
To study Rosmarinus officinalis (Rosemary) essential oil effect on primary hypotension and its influence on both physical and psychological aspects responsible for health-related quality of life (HRQOL) of patients. Thirty-two patients with diagnosed hypotension were recruited between March 2007 and September 2008 for a prospective study for 72 weeks in a Spanish pharmacy. Clinical evaluation was carried out through the control of systolic and diastolic blood pressure levels (SBP and DBP, respectively) according to the International Standards from the American Society of Hypertension. HRQOL data were recorded within the SF-36 Health Survey(®) questionnaire throughout the study. Statistical methods were used as the essential tools to evaluate the effectiveness of Rosemary essential oil and to assess the relationship between the two quantitative variables (SBP and DBP) and scores from physical and mental summary components (PSC and MSC) obtained from the SF-36 Health Survey. Both blood pressure variables of SBP and DBP reflect the clinically significant antihypotensive effect of Rosemary essential oil that was maintained throughout the treatment period. After validation of the use of the questionnaire (Cronbach's alpha coefficient>0.82), statistically significant differences have been found between pre-treatment and post-treatment values of PSC and MSC, which indicate an improvement in these parameters that is directly related to the variation in blood pressure values. The increase achieved in blood pressure values after administration of Rosemary essential oil is clinically significant. The results obtained from this prospective clinical trial prove the effectiveness of statistical methodology as a new approach to explain the antihypotensive effect of rosemary essential oil and its relationship with the improvement in patients' quality of life.


Nutritional supplements with intravenously administered lipid, protein hydrolysate, and glucose in small premature infants.
Nutritional support with intravenously administered fat, protein hydrolysate, and glucose was given to 23 premature infants weighing less than 1,500 gm at birth. Growth in the 19 survivors approximated intrauterine growth rates in all dimensions. Complications included sodium imbalance and transient intolerance of the smallest and sickest infants to all components: carbohydrate, protein, fat, and even water. Autopsies in four patients who died did not reveal any untoward findings clearly related to the use of intravenously given lipids. Intravenously given fat has a potentially useful role in the nutritional support of small premature infants.


Nanotechnological Advances in the Treatment of Epilepsy.
Epilepsy is known as one of the major challenges for medical science. The sudden appearance of a seizure has been a significant health emergency as it may lead to further complications. Although key advancements have been achieved in terms of pharmacological approaches for epilepsy, many issues remain uncertain. Lipid carriers have been at the forefront, especially in neurodegenerative diseases, such as epilepsy, Alzheimer's, dementia, etc. The blood-brain barrier still appears to be a major impediment in the successful treatment of epileptic seizures. This is mainly due to the limited bioavailability of most anti-convulsant drugs. The present review encompasses the issues underlying the current approach for epilepsy drug treatment and highlights the newer, novel, and more precise drug delivery system to manage seizures. The advantage of using a lipidbased delivery system is its superior absorption in the brain cells. Ample evidence shows that reducing the particle size also infuses the drug easily through the blood-brain barrier. The application of liposomes, polymeric nanoparticles, metallic nanoparticles, and solid lipid nanoparticles for the treatment and management of epilepsy has been highlighted in the present review. This review provides an overview of the current status of the treatment and recent advances in the treatment of epilepsy.


Performance-Enhancing Drugs I: Understanding the Basics of Testing for Banned Substances.
Whenever athletes willfully or accidentally ingest performance-enhancing drugs or other banned substances (such as drugs of abuse), markers of those drugs can be detected in biological samples (e.g., biofluids: urine, saliva, blood); in the case of some drugs, that evidence can be apparent for many weeks following the last exposure to the drug. In addition to the willful use of prohibited drugs, athletes can accidentally ingest banned substances in contaminated dietary supplements or foods and inadvertently fail a drug test that could mean the end of an athletic career and the loss of a good reputation. The proliferation of performance-enhancing drugs and methods has required a corresponding increase in the analytical tools and methods required to identify the presence of banned substances in biofluids. Even though extraordinary steps have been taken by organizations such as the World Anti-Doping Agency to limit the use of prohibited substances and methods by athletes willing to cheat, it is apparent that some athletes continue to avoid detection by using alternative doping regimens or taking advantage of the limitations in testing methodologies. This article reviews the testing standards and analytical techniques underlying the procedures used to identify banned substances in biological samples, setting the stage for future summaries of the testing required to establish the use of steroids, stimulants, diuretics, and other prohibited substances.


[Characteristics of the capsular antigen preparations isolated from a broth culture of Yersinia pestis strain EV].
The physico-chemical and immunobiological study of the preparation of capsular antigen, isolated by a high-yield method from the culture fluid of Y. pestis grown in nutrient broth at 37 degrees C and practically free from the admixture of polysaccharide and nucleic acids, has been made. The study has revealed that this capsular antigen is protein with a molecular weight of 1.65-1.69 X 10(6) D and sedimentation constant equal to 13.4S; its viscosity is 220 ml/g and its isoelectric point is at pH 4.1. The preparations have been found to possess immunogenic and allergenic properties and to be capable of sensitizing formulated, tannin-treated sheep red blood cells to plague antiserum.


Feasibility and safety of mass drug coadministration with azithromycin and ivermectin for the control of neglected tropical diseases: a single-arm intervention trial.
Mass drug administration has made a major contribution to the public health control of several important neglected tropical diseases. For settings with more than one endemic disease, combined mass drug administration has potential practical advantages compared with separate programmes but needs confirmation of feasibility and safety. We undertook a study of mass drug administration in the Solomon Islands for trachoma and scabies control using ivermectin and azithromycin, key drugs in the control of neglected tropical diseases worldwide. The entire population of Choiseul province, Solomon Islands, was eligible to participate. An azithromycin-based mass drug administration regimen was offered in line with standard recommendations for trachoma elimination (oral azithromycin or topical tetracycline). An ivermectin-based mass drug administration regimen was offered at the same time (oral ivermectin or topical permethrin), with a further dose 7-14 days later, using a modified version of a regimen demonstrated to be effective for scabies control. All participants underwent safety assessments 7-14 days later. Participants in ten randomly selected sentinel villages underwent a more detailed safety assessment. Routine health system reports of hospital or clinic admissions and deaths were also obtained to compare health outcomes in the 12 month period before and after the mass drug administration. The study enrolled 26 188 participants, 99·3% of the estimated resident population as determined at the 2009 census. Of those enrolled, 25 717 (98·2%) received the trachoma regimen and 25 819 (98·6%) received the first dose of the scabies regimen between Sept 1, and Oct 2, 2015. A second dose of the scabies regimen was received by 21 931 (83·7%) of participants. Adverse events, all mild and transient, were recorded in 571 (2·6%) of the entire study population and 58 (4·1%) of participants in the ten sentinel villages. In the 12 months before and after the mass drug administration the numbers of hospital admissions (1530 vs 1602) and deaths (73 vs 83) were similar. In the month after the mass drug administration, 84 individuals were admitted to hospital and two died, compared with a monthly median of 116 admissions (IQR 106-159) and six deaths (IQR 4-7) in the 12 months before and after the mass drug administration. In the largest trial so far involving coadministration of regimens based on ivermectin and azithromycin, the combination was safe and feasible in a population of more than 26 000 people. Coadministration of mass drug administration based on these two drugs opens up new potential for the control of neglected tropical diseases. International Trachoma Initiative, Murdoch Children's Research Institute, Scobie and Claire Mackinnon Trust, Wellcome Trust.


Geographic variations in coronary heart disease. A perspective on risk factors and prevention.
Mankind's greatest epidemic: heart disease--ischemic heart disease or coronary heart disease (CHD)--has reached epidemic proportions striking more and more at younger subjects. It will result in coming years, in the greatest epidemic mankind has faced unless we are able to reverse the trend by concentrated research into its causes and prevention. Under the auspices of the World Health Organization the [executive] board expressed a wish that countries most affected by cardiovascular diseases increase their efforts, both to set up efficient services for control and to carry out more extensive research programs (20).


Creating HIV risk profiles for men in South Africa: a latent class approach using cross-sectional survey data.
Engaging at-risk men in HIV prevention programs and services is a current priority, yet there are few effective ways to identify which men are at highest risk or how to best reach them. In this study we generated multi-factor profiles of HIV acquisition/transmission risk for men in Durban, South Africa, to help inform targeted programming and service delivery. Data come from surveys with 947 men ages 20 to 40 conducted in two informal settlements from May to September 2017. Using latent class analysis (LCA), which detects a small set of underlying groups based on multiple dimensions, we identified classes based on nine HIV risk factors and socio-demographic characteristics. We then compared HIV service use between the classes. We identified four latent classes, with good model fit statistics. The older high-risk class (20% of the sample; mean age 36) were more likely married/cohabiting and employed, with multiple sexual partners, substantial age-disparity with partners (eight years younger on-average), transactional relationships (including more resource-intensive forms like paying for partner's rent), and hazardous drinking. The younger high-risk class (24%; mean age 27) were likely unmarried and employed, with the highest probability of multiple partners in the last year (including 42% with 5+ partners), transactional relationships (less resource-intensive, e.g., clothes/transportation), hazardous drinking, and inequitable gender views. The younger moderate-risk class (36%; mean age 23) were most likely unmarried, unemployed technical college/university students/graduates. They had a relatively high probability of multiple partners and transactional relationships (less resource-intensive), and moderate hazardous drinking. Finally, the older low-risk class (20%; mean age 29) were more likely married/cohabiting, employed, and highly gender-equitable, with few partners and limited transactional relationships. Circumcision (status) was higher among the younger moderate-risk class than either high-risk class (p < 0.001). HIV testing and treatment literacy score were suboptimal and did not differ across classes. Distinct HIV risk profiles among men were identified. Interventions should focus on reaching the highest-risk profiles who, despite their elevated risk, were less or no more likely than the lower-risk to use HIV services. By enabling a more synergistic understanding of subgroups, LCA has potential to enable more strategic, data-driven programming and evaluation.


Interaction of heterogeneous nuclear ribonucleoprotein C1/C2 with a novel cis-regulatory element within p53 mRNA as a response to cytostatic drug treatment.
We describe a novel cis-element in the 5' coding region of p53 mRNA and its interaction with heterogeneous nuclear ribonucleoprotein (hnRNP)C1/C2. This element is located in a putative hairpin loop structure, within the first 101 nucleotides downstream of the start codon. The binding of hnRNPC1/C2 is strongly enhanced in response to the DNA-damaging drug cisplatin [cis-diamminedichloroplatinum(II)] and the cytostatic transcriptional inhibitor actinomycin D (dactinomycin), both known inducers of apoptosis and p53. Strongly stimulated binding is observed in both nuclear and cytoplasmic compartments, and it is accompanied by a cytoplasmic increase of hnRNPC1/C2. Changes in hnRNPC1/C2 protein levels are not proportional to binding activity, suggesting qualitative changes in hnRNPC1/C2 upon activation. Phosphorylation studies reveal contrasting characteristics of the cytoplasmic and nuclear hnRNPC1/C2 interaction with p53 mRNA. Results from chimeric p53-luciferase reporter constructs suggest that hnRNPC1/C2 regulates p53 expression via this binding site. Our results are consistent with a mechanism in which the interaction of hnRNPC1/C2 with a cis-element within the coding region of the p53 transcript regulates the expression of p53 mRNA before and during apoptosis. In addition, we report that preapoptotic signals induced by transcriptional inhibition trigger the appearance of a truncated, exclusively cytoplasmic 43-kDa variant of p53 before apoptosis.


Inhibitory effects of condensed tannins on angiotensin converting enzyme.
Effects of condensed tannins isolated from Rhei Rhizoma on the activities of angiotensin converting enzyme (ACE) and various proteases were examined in vitro. Among the various condensed tannins tested, procyanidin B-5 3,3'-di-O-gallate and procyanidin C-1 3,3',3"-tri-O-gallate strongly inhibited the activity of ACE. The concentration of procyanidin B-5 3,3'-di-O-gallate required for 50% inhibition of ACE was 1.3 X 10(-6) M. The inhibition of ACE by condensed tannins was reversible and non-competitive, according to dialysis and to Dixon plots. However, over one hundred times the concentration was required to inhibit activities of other proteases such as trypsin, chymotrypsin, leucine aminopeptidase, carboxypeptidase A and urinary kallikrein. These results suggest that the inhibitory effects of condensed tannins on the activities of ACE are specific.


Risk stratification in acute pulmonary embolism: frequency and impact on treatment decisions and outcomes.
Guidelines have recommended that risk stratification be performed in patients diagnosed with an acute pulmonary embolism (PE). No study has described the use of risk stratification in routine clinical practice. The purpose of this study was to measure the frequency and impact of risk stratification on treatment decisions and outcomes in patients admitted with acute PE. A retrospective cohort study was conducted of all of the patients admitted with acute PE at two Geisinger community-based teaching hospitals between 2006 and 2011. Baseline demographics, vital signs, and relevant clinical variables were recorded. The Pulmonary Embolism Severity Index was calculated for each patient. Risk stratification was defined as the measurement of either a biomarker or an echocardiogram within 24 hours of admission. The outcomes measured were short-term adverse events (in-hospital mortality or need for intensive care) and 30-day mortality. The mean age for the study cohort (n = 889) was 61 ± 17 years and 52% were men. Overall, 59% of study subjects were risk stratified. The frequency of risk stratification did not change over time. Risk stratification was associated with assignment to a higher acuity of care and increased use of thrombolysis and inferior vena cava filter placement. When controlling for severity of illness, risk stratification was a significant predictor of worsened short-term adverse outcome (odds ratio 3.43, 95% confidence interval 1.75-6.74, P < 0.001) but was not associated with improved 30-day mortality (odds ratio 1.14, 95% confidence interval 0.66-1.95, P = 0.64). Risk stratification is frequently performed in patients admitted with acute PE and has had a stable prevalence during a 5-year period. The use of risk stratification in acute PE is associated with assignment to higher levels of care and with more advanced treatments. Despite more intense treatment, risk stratification does not improve either short-term outcomes or 30-day mortality.


Glutamine-181 is crucial in the enzymatic activity and substrate specificity of human endoplasmic-reticulum aminopeptidase-1.
ERAP-1 (endoplasmic-reticulum aminopeptidase-1) is a multifunctional enzyme with roles in the regulation of blood pressure, angiogenesis and the presentation of antigens to MHC class I molecules. Whereas the enzyme shows restricted specificity toward synthetic substrates, its substrate specificity toward natural peptides is rather broad. Because of the pathophysiological significance of ERAP-1, it is important to elucidate the molecular basis of its enzymatic action. In the present study we used site-directed mutagenesis to identify residues affecting the substrate specificity of human ERAP-1 and identified Gln(181) as important for enzymatic activity and substrate specificity. Replacement of Gln(181) by aspartic acid resulted in a significant change in substrate specificity, with Q181D ERAP-1 showing a preference for basic amino acids. In addition, Q181D ERAP-1 cleaved natural peptides possessing a basic amino acid at the N-terminal end more efficiently than did the wild-type enzyme, whereas its cleavage of peptides with a non-basic amino acid was significantly reduced. Another mutant enzyme, Q181E, also revealed some preference for peptides with a basic N-terminal amino acid, although it had little hydrolytic activity toward the synthetic peptides tested. Other mutant enzymes, including Q181N and Q181A ERAP-1s, revealed little enzymatic activity toward synthetic or peptide substrates. These results indicate that Gln(181) is critical for the enzymatic activity and substrate specificity of ERAP-1.