Effects of genetic variation in H3K79 methylation regulatory genes on clinical blood pressure and blood pressure response to hydrochlorothiazide.
Nearly one-third of the United States adult population suffers from hypertension. Hydrochlorothiazide (HCTZ), one of the most commonly used medications to treat hypertension, has variable efficacy. The renal epithelial sodium channel (ENaC) provides a mechanism for fine-tuning sodium excretion, and is a major regulator of blood pressure homeostasis. DOT1L, MLLT3, SIRT1, and SGK1 encode genes in a pathway that controls methylation of the histone H3 globular domain at lysine 79 (H3K79), thereby modulating expression of the ENaCα subunit. This study aimed to determine the role of variation in these regulatory genes on blood pressure response to HCTZ, and secondarily, untreated blood pressure. We investigated associations between genetic variations in this candidate pathway and HCTZ blood pressure response in two separate hypertensive cohorts (clinicaltrials.gov NCT00246519 and NCT00005520). In a secondary, exploratory analysis, we measured associations between these same genetic variations and untreated blood pressure. Associations were measured by linear regression, with only associations with P ≤ 0.01 in one cohort and replication by P ≤ 0.05 in the other cohort considered significant. In one cohort, a polymorphism in DOT1L (rs2269879) was strongly associated with greater systolic (P = 0.0002) and diastolic (P = 0.0016) blood pressure response to hydrochlorothiazide in Caucasians. However, this association was not replicated in the other cohort. When untreated blood pressure levels were analyzed, we found directionally similar associations between a polymorphism in MLLT3 (rs12350051) and greater untreated systolic (P < 0.01 in both cohorts) and diastolic (P < 0.05 in both cohorts) blood pressure levels in both cohorts. However, when further replication was attempted in a third hypertensive cohort and in smaller, normotensive samples, significant associations were not observed. Our data suggest polymorphisms in DOT1L, MLLT3, SIRT1, and SGK1 are not likely associated with blood pressure response to HCTZ. However, a possibility exists that rs2269879 in DOT1L could be associated with HCTZ response in Caucasians. Additionally, exploratory analyses suggest rs12350051 in MLLT3 may be associated with untreated blood pressure in African-Americans. Replication efforts are needed to verify roles for these polymorphisms in human blood pressure regulation.


Escherichia coli K1 promotes the ligation of CD47 with thrombospondin-1 to prevent the maturation of dendritic cells in the pathogenesis of neonatal meningitis.
Dendritic cells (DCs) are professional APCs providing a critical link between adaptive and innate immune responses. Our previous studies have shown that Escherichia coli K1 internalization of myeloid DCs suppressed the maturation of the cells for which outer membrane protein A (OmpA) expression is essential. In this study, we demonstrate that infection of DCs with OmpA(+) E. coli significantly upregulates the expression of CD47, an integrin-associated protein, and its natural ligand thrombospondin 1 (TSP-1). Pretreatment of DCs with anti-CD47 blocking Ab or knocking down the expression of CD47 or TSP-1, but not signal regulatory protein alpha by small interfering RNA, abrogated the suppressive effect of E. coli K1. Ligation of CD47 with a mAb prevented the maturation and cytokine production by DCs upon stimulation with LPS similar to the inhibitory effect induced by OmpA(+) E. coli. In agreement with the in vitro studies, suppression of CD47 or TSP-1 expression in newborn mice by a novel in vivo small interfering RNA technique protected the animals against E. coli K1 meningitis. Reconstitution of CD47 knockdown mice with CD47(+) DCs renders the animals susceptible to meningitis by E. coli K1, substantiating the role of CD47 expression in DCs for the occurrence of meningitis. Our results demonstrate a role for CD47 for the first time in bacterial pathogenesis and may be a novel target for designing preventive approaches for E. coli K1 meningitis.


Complete complex conjugate resolved heterodyne swept source optical coherence tomography using a dispersive optical delay line: erratum.
We correct an error in our previous paper [Biomed. Opt. Express 2, 1218 (2011)] which led to an erroneous conclusion that a dispersive optical delay line (DODL) used in a swept source optical coherence tomography (SSOCT) system generated a pure phase modulation allowing for complex conjugate artifact removal in Fourier domain OCT via optical heterodyning. We now understand that an alternate phenomenon known as coherence revival was responsible for the observed phase modulation, while the DODL provided a compact means of generating a large group delay with readily adjustable group velocity dispersion compensation.


C-terminal heat shock protein 90 modulators produce desirable oncogenic properties.
The cellular protection mechanism, the heat shock response, is only activated by classical heat shock 90 inhibitors (Hsp90) that "target" the N-terminus of the protein, but not by those that modulate the C-terminus. Significant differences in cytotoxicity (nanomolar) for classical inhibitors versus their ability to modulate Hsp90 (low micromolar) are discussed. In contrast, molecules that modulate Hsp90's C-terminus show similar IC50 values for cytotoxicity and Hsp90 inhibition. A comparison between the two types of Hsp90 inhibitors suggests that classical inhibitors may be modulating an alternative biological target that stresses the cell rather directly inhibiting Hsp90, whereas C-terminal modulators are most likely acting by directly inhibiting Hsp90.


The State of Federal Research Funding in Genetics as Reflected by Members of the Genetics Society of America.
Scientific progress runs on the intellect, curiosity, and passion of its practitioners fueled by the research dollars of its sponsors. The concern over research funding in biology in general and genetics in particular led us to survey the membership of the Genetics Society of America for information about the federal support of genetics at the level of individual principal investigators. The results paint a mosaic of circumstances-some good, others not so good-that describes some of our present challenges with sufficient detail to suggest useful steps that could address the challenges.


Inotuzumab ozogamicin murine analog-mediated B-cell depletion reduces anti-islet allo- and autoimmune responses.
B cells participate in the priming of the allo- and autoimmune responses, and their depletion can thus be advantageous for islet transplantation. Herein, we provide an extensive study of the effect of B-cell depletion in murine models of islet transplantation. Islet transplantation was performed in hyperglycemic B-cell-deficient(μMT) mice, in a purely alloimmune setting (BALB/c into hyperglycemic C57BL/6), in a purely autoimmune setting (NOD.SCID into hyperglycemic NOD), and in a mixed allo-/autoimmune setting (BALB/c into hyperglycemic NOD). Inotuzumab ozogamicin murine analog (anti-CD22 monoclonal antibody conjugated with calicheamicin [anti-CD22/cal]) efficiently depleted B cells in all three models of islet transplantation examined. Islet graft survival was significantly prolonged in B-cell-depleted mice compared with control groups in transplants of islets from BALB/c into C57BL/6 (mean survival time [MST]: 16.5 vs. 12.0 days; P = 0.004), from NOD.SCID into NOD (MST: 23.5 vs. 14.0 days; P = 0.03), and from BALB/c into NOD (MST: 12.0 vs. 5.5 days; P = 0.003). In the BALB/c into B-cell-deficient mice model, islet survival was prolonged as well (MST: μMT = 32.5 vs. WT = 14 days; P = 0.002). Pathology revealed reduced CD3(+) cell islet infiltration and confirmed the absence of B cells in treated mice. Mechanistically, effector T cells were reduced in number, concomitant with a peripheral Th2 profile skewing and ex vivo recipient hyporesponsiveness toward donor-derived antigen as well as islet autoantigens. Finally, an anti-CD22/cal and CTLA4-Ig-based combination therapy displayed remarkable prolongation of graft survival in the stringent model of islet transplantation (BALB/c into NOD). Anti-CD22/cal-mediated B-cell depletion promotes the reduction of the anti-islet immune response in various models of islet transplantation.


Microlaparoscopic-assisted lumboperitoneal shunt in the lateral position for pseudotumor cerebri in a morbidly obese adolescent.
Pseudotumor cerebri or idiopathic intracranial hypertension is a known complication of morbid obesity that often requires neurosurgical intervention for worsening symptoms. Placement of a lumboperitoneal shunt (LPS) is one of the treatment options, but in a morbidly obese patient it can be technically challenging. We describe the use of 3-mm instrumentation for assistance in placing the peritoneal end of the shunt. A 16-year-old morbidly obese girl with a diagnosis of pseudotumor cerebri and decreasing visual acuity and contraction of her visual fields underwent lumboperitoneal shunt placement. Due to her body habitus secondary to her morbid obesity a microlaparoscopic-assisted approach was utilized for placement of the peritoneal end of the lumboperitoneal shunt. No operative or postoperative problems occurred, and she was discharged home with resolution of symptoms. Her visual acuity and fields had normalized at 3-month follow-up. Microlaparoscopic-assisted lumboperitoneal shunt placement in the lateral position is an efficient and safe method for the treatment of pseudotumor cerebri. It is a minimally invasive, simple, effective tool for placing the peritoneal catheter for LPS.


Mutualistic mimicry enhances species diversification through spatial segregation and extension of the ecological niche space.
Species richness varies among clades, yet the drivers of diversification creating this variation remain poorly understood. While abiotic factors likely drive some of the variation in species richness, ecological interactions may also contribute. Here, we examine one class of potential contributors to species richness variation that is particularly poorly understood: mutualistic interactions. We aim to elucidate large-scale patterns of diversification mediated by mutualistic interactions using a spatially explicit population-based model. We focus on mutualistic Müllerian mimicry between conspicuous toxic prey species, where convergence in color patterns emerges from predators' learning process. To investigate the effects of Müllerian mimicry on species diversification, we assume that some speciation events stem from shifts in ecological niches, and can also be associated with shift in mimetic color pattern. Through the emergence of spatial mosaics of mimetic color patterns, Müllerian mimicry constrains the geographical distribution of species and allows different species occupying similar ecological niches to exist simultaneously in different regions. Müllerian mimicry and the resulting spatial segregation of mimetic color patterns thus generate more balanced phylogenetic trees and increase overall species diversity. Our study sheds light on complex effects of Müllerian mimicry on ecological, spatial, and phylogenetic diversification.


Composite ligand shells on gold nanoprisms - an ensemble and single particle study.
The attachment of thiolated molecules onto gold surfaces is one of the most extensively used and robust ligand exchange approaches to exploit the nanooptical features of nanoscale and nanostructured plasmonic materials. In this work, the impact of thiol adsorption on the optical properties of wet-chemically synthesized gold nanoprisms is studied both at the ensemble and single particle level to investigate the build-up of more complex ligand layers. Two prototypical ligands with different lengths have been investigated ((16-mercaptohexadecyl)trimethylammonium bromide - MTAB and thiolated polyethylene glycol - mPEG-SH). From ensemble experiments it is found that composite ligand layers are obtained by the sequential addition of the two thiols, and an island-like surface accumulation of the molecules can be anticipated. The single particle experiment derived chemical interface damping and resonance energy changes further support this and show additionally that when the two thiols are used simultaneously, a higher density, intermixed layer is formed. Hence, when working with more than a single type of ligand during surface modification, sequential adsorption is preferred for the combination of accessible essential surface functionalities, whereas for high overall loading the simultaneous use of the different ligand types is favourable.


A small variation in diet influences the Lactobacillus strain composition in the crop of broiler chickens.
Feed composition has the potential to influence the activities of bacteria that colonize the digestive tract of broiler chickens with important consequences for animal health, well being, and food safety. In this study, the gut microbiota of two groups of broiler chickens raised in immediate vicinity but fed either a standard corn/soybean meal ration (corn-soy, CS) or a ration high in wheat middlings (high wheat, HW) was characterized. The findings revealed that this small variation in feed composition did not influence the distribution of microbial species present in the microbial community throughout the digestive tract. However, diet variation markedly influenced the Lactobacillus strain composition in the crop. Most striking, the dominant type in birds on the CS diet (Lactobacillus agilis type R5), which comprised 25% of the isolates, was not detected in birds fed the HW diet. The latter birds harbored a different strain of L. agilis (type R1) in a significantly higher ratio than birds on the CS diet. Several other strains were also specific to the particular diet. In conclusion, this study showed that a small variation in the composition of chicken feed that does not result in detectable differences in species composition can still have an impact on which microbial strains become dominant in the digestive tract. This finding has relevance in the application of probiotics and other direct-fed microbials in poultry husbandry.


Impact of pathological T3a upstaging on oncological outcomes of clinical T1 renal cell carcinoma: a meta-analysis.
Objective: The study aims to assess the prognostic impact of pathological T3a upstaging in clinical T1 renal cell carcinoma (RCC) on clinical outcomes. Methods: We performed a systematic literature search of PMC, Embase, Web of Science, and Cochrane library from inception to April 2019 for studies that investigated the prognostic significance of pathological T3a upstaging in clinical T1 RCC after surgery and conducted a standard meta-analysis on survival outcomes. Results: Overall, nine studies including 101,505 clinical T1 RCC patients were identified, in which 5,560 (5.5%) patients were upstaged to T3a after surgical treatment. Meta-analysis results showed that pT3a upstaging from clinical T1 RCC was significantly associated with poor recurrence-free survival (RFS; pooled hazard ratio [HR] 2.16, 95% confidence interval [CI] 1.70-2.75; P<0.001), overall survival (OS; pooled HR 1.36, 95% CI 1.24-1.50; P<0.001), and cancer-specific survival (CSS; pooled HR 2.11, 95% CI 1.58-2.83; P<0.001). Subgroup analyses by surgical type demonstrated that pT3a upstaging remains a significant prognostic factor for RFS and OS in RCC patients who underwent different surgical treatments. Conclusions: Current available evidence strongly supported that postoperative pT3a upstaging has a significant negative impact on RFS, OS, and CSS in clinical T1 RCC patients. Clinical T1 RCC patients with pT3a upstaging after surgery should be closely monitored by clinician and should receive close follow-up for their poor prognosis.


Therapeutic Options for Recurrence of Primary Focal Segmental Glomerulonephritis (FSGS) in the Renal Allograft: Single-Center Experience.
Focal Segmental Glomerulosclerosis (FSGS) recurrence after kidney transplantation (KTx) is relatively frequent and is associated with poor graft survival. The aim of this study was to investigate which management strategies were associated with better outcomes in our cohort of KTx recipients with primary FSGS. We retrospectively collected data on patients with primary FSGS who received a KTx between 1993 and 2019. A history of biopsy proven FSGS in native kidneys and new onset of significant proteinuria early post-KTx led to the diagnosis of recurrence, which was confirmed by graft biopsy. From 1993 to 2019 we performed 46 KTxs in patients with primary FSGS. We identified 26 episodes of recurrence in 25 patients, 67% of them occurring in males. They were younger at the time of KTx (33.8 vs. 41.1 years old, p = 0.067) and had progressed to end stage renal disease (ESRD) faster after FSGS diagnosis (61.4 vs. 111.2 months, p = 0.038), while they were less likely to have received prophylactic plasmapheresis (61.5% vs. 90%, p = 0.029). 76.7% of recurrences were found early, after a median of 0.5 months (IQR 0.1-1) with a median proteinuria was 8.5 (IQR 4.9-11.9) g/day. All patients with recurrence were treated with plasmapheresis, while 8 (30.7%) additionally received rituximab, 1 (3.8%) abatacept, and 4 (15.4%) ACTH. 7 (27%) patients experienced complete and 11 (42.3%) partial remission after a mean time of 3 (±1.79) and 4.4 (±2.25) months, respectively. Prognosis was worse for patients who experienced a recurrence. Eleven (42.3%) patients lost their graft from FSGS in a median time of 33 (IQR 17.5-43.3) months. In this series of patients, primary FSGS recurred frequently after KTx. Prophylacic plasmapheresis was shown efficacious in avoiding FSGS recurrence, while timely diagnosis and plasmapheresis-based regimens induced remission in more than half of the patients.


Role of the renin-angiotensin system in cardiac hypertrophy induced in rats by hyperthyroidism.
This study was conducted to examine whether the renin-angiotensin system contributes to hyperthyroidism-induced cardiac hypertrophy without involving the sympathetic nervous system. Sprague-Dawley rats were divided into control-innervated, control-denervated, hyperthyroid-innervated, and hyperthyroid-denervated groups using intraperitoneal injections of thyroxine and 6-hydroxydopamine. After 8 wk, the heart-to-body weight ratio increased in hyperthyroid groups (63%), and this increase was only partially inhibited by sympathetic denervation. Radioimmunoassays and reverse transcription-polymerase chain reaction revealed increased cardiac levels of renin (33%) and angiotensin II (53%) and enhanced cardiac expression of renin mRNA (225%) in the hyperthyroid groups. These increases were unaffected by sympathetic denervation or 24-h bilateral nephrectomy. In addition, losartan and nicardipine decreased systolic blood pressure to the same extent, but only losartan caused regression of thyroxine-induced cardiac hypertrophy. These results suggest that thyroid hormone activates the cardiac renin-angiotensin system without involving the sympathetic nervous system or the circulating renin-angiotensin system; the activated renin-angiotensin system contributes to cardiac hypertrophy in hyperthyroidism.


Differences in physical environmental characteristics between adolescents' actual and shortest cycling routes: a study using a Google Street View-based audit.
The objective evaluation of the physical environmental characteristics (e.g. speed limit, cycling infrastructure) along adolescents' actual cycling routes remains understudied, although it may provide important insights into why adolescents prefer one cycling route over another. The present study aims to gain insight into the physical environmental characteristics determining the route choice of adolescent cyclists by comparing differences in physical environmental characteristics between their actual cycling routes and the shortest possible cycling routes. Adolescents (n = 204; 46.5% boys; 14.4 ± 1.2 years) recruited at secondary schools in and around Ghent (city in Flanders, northern part of Belgium) were instructed to wear a Global Positioning System device in order to identify cycling trips. For all identified cycling trips, the shortest possible route that could have been taken was calculated. Actual cycling routes that were not the shortest possible cycling routes were divided into street segments. Segments were audited with a Google Street View-based tool to assess physical environmental characteristics along actual and shortest cycling routes. Out of 160 actual cycling trips, 73.1% did not differ from the shortest possible cycling route. For actual cycling routes that were not the shortest cycling route, a speed limit of 30 km/h, roads having few buildings with windows on the street side and roads without cycle lane were more frequently present compared to the shortest possible cycling routes. A mixed land use, roads with commercial destinations, arterial roads, cycle lanes separated from traffic by white lines, small cycle lanes and cycle lanes covered by lighting were less frequently present along actual cycling routes compared to the shortest possible cycling routes. Results showed that distance mainly determines the route along which adolescents cycle. In addition, adolescents cycled more along residential streets (even if no cycle lane was present) and less along busy, arterial roads. Local authorities should provide shortcuts free from motorised traffic to meet adolescents' preference to cycle along the shortest route and to avoid cycling along arterial roads.


[Atherosclerotic renal artery stenosis].
The most common cause of obstructive renal artery disease is atherosclerosis, accounting for 90 % of cases of renal artery stenosis. Atherosclerotic renal artery stenosis can be associated with renovascular hypertension, ischemic nephropathy, or both or it may occur alone. The prevalence of atherosclerotic renal artery stenosis among hypertensive patients is estimated between 1 and 5 %, but the frequency rises among patients with refractory hypertension (20 %) coronary heart disease (15 to 20 %) or peripheral arterial disease (30 to 40 %). The gold standard for diagnosing renal artery disease is contrast renal arteriography. MR angiography, CT angiography and color duplex ultrasonography have the highest sensitivity and specifity among the non invasive screening methods. Therapy is based on consequent medical treatment of hypertension, antiplatelet therapy and modification of risk factors for atherosclerosis. Revascularisation is advised in patients with severe hypertension, in patients with pulmonary edema and cases of acute worsening of renal function. Percutaneous angioplasty with stent implantation is the method of choice for revascularisation. The prognosis of patients with atherosclerotic renal artery stenosis is determined by cardiovascular and renal complications.


On the probability of matching DNA fingerprints.
Forensic scientists commonly assume that DNA fingerprint patterns are infrequent in the general population and that genotypes are independent across loci. To test these assumptions, the number of matching DNA patterns in two large databases from the Federal Bureau of Investigation (FBI) and from Lifecodes was determined. No deviation from independence across loci in either database was apparent. For the Lifecodes database, the probability of a three-locus match ranges from 1 in 6,233 in Caucasians to 1 in 119,889 in Blacks. When considering all trios of five loci in the FBI database, there was only a single match observed out of more than 7.6 million comparisons. If independence is assumed, the probability of a five-locus match ranged from 1.32 x 10(-12) in Southeast Hispanics to 5.59 x 10(-14) in Blacks, implying that the minimum number of possible patterns for each ethnic group is several orders of magnitude greater than their corresponding population sizes in the United States. The most common five-locus pattern can have a frequency no greater than about 10(-6). Hence, individual five-locus DNA profiles are extremely uncommon, if not unique.


In vitro antiplasmodial activities and synergistic combinations of differential solvent extracts of the polyherbal product, Nefang.
Nefang, a polyherbal product composed of Mangifera indica (bark and leaf), Psidium guajava, Carica papaya, Cymbopogon citratus, Citrus sinensis, and Ocimum gratissimum (leaves), is a potential therapy against P. falciparum malaria. In vitro antiplasmodial activities of its constituent solvent extracts were analyzed on CQ-sensitive (3D7) and multidrug resistant (Dd2) P. falciparum strains. The interactions involving the differential solvent extracts were further analyzed using a variable potency ratio drug combination approach. Effective concentration 50 (EC50) values were determined by nonlinear regression curve-fitting of the dose-response data and used in calculating the fractional inhibitory concentration 50 (FIC50) and combination indices (CI) for each pair. The derived EC50 values (3D7/Dd2, μ g/mL) are Nefang-96.96/55.08, MiB-65.33/34.58, MiL-82.56/40.04, Pg-47.02/25.79, Cp-1188/317.5, Cc-723.3/141, Cs-184.4/105.1, and Og-778.5/118.9. Synergism was obtained with MiB/Pg (CI = 0.351), MiL/Pg (0.358), MiB/Cs (0.366), MiL/Cs (0.482), Pg/Cs (0.483), and Cs/Og (0.414) when analyzed at equipotency ratios. Cytotoxicity testing of Nefang and the solvent extracts on two human cell lines (Hep G2 and U2OS) revealed no significant toxicity relative to their antiplasmodial activities (SI > 20). Taken together, our data confirm the antimalarial activities of Nefang and its constituent plant extracts and identified extract pairs with promising synergistic interactions for exploitation towards a rational phytotherapeutic and evidence-based antimalarial drug discovery.


beta-Glucuronidase from Escherichia coli as a gene-fusion marker.
We have developed a gene-fusion system based on the Escherichia coli beta-glucuronidase gene (uidA). The uidA gene has been cloned from E. coli K-12 and its entire nucleotide sequence has been determined. beta-Glucuronidase has been purified to homogeneity and characterized. The enzyme has a subunit molecular weight of 68,200, is very stable, and is easily and sensitively assayed using commercially available substrates. We have constructed gene fusions of the E. coli lacZ promoter and coding region with the coding region of the uidA gene that show beta-glucuronidase activity under lac control. Plasmid vectors have been constructed to facilitate the transfer of the beta-glucuronidase coding region to heterologous control regions, using many different restriction endonuclease cleavage sites. There are several biological systems in which uidA-encoded beta-glucuronidase may be an attractive alternative or complement to previously described gene-fusion markers such as beta-galactosidase or chloramphenicol acetyltransferase.


Potencies and channel properties induced by semirigid agonists at frog nicotinic acetylcholine receptors.
Structure-activity relationships were investigated in a series of semirigid nicotinic agonists. Three of the agonists, (-)-ferruginine methiodide, arecoline methiodide, and its ketonic analogue arecolone methiodide, were cyclic analogues of anatoxin-a, a potent, naturally occurring, bicyclic alkaloid. Two other cyclic agonists, (-)-cytisine and (+/-)-muscarone, and the simplest agonist, the tetramethylammonium ion, were also tested. Arecolone methiodide and (-)-ferruginine methiodide have been tested as nicotinic agonists for the first time. Relative potency was assayed by contracture on the rectus abdominis muscle of the frog Rana pipiens. Natural, (+)-anatoxin-a, the most active of all of the agonists, was more than twice as potent as racemic anatoxin-a. Arecolone methiodide ranked after anatoxin-a in potency, being 8.6 times more potent than carbamycholine. A correlation between nicotinic potency and steric requirements probably involves the position of positively charged groups out of the plane defined by the carbonyl group and its two substituents. Channel properties induced by the agonists were evaluated by Fourier analysis of the end-plate current noise that resulted when the agonists were iontophoretically applied to frog sartorius muscle fibers. Average channel lifetimes were exponential functions of membrane potential, but the voltage sensitivity of channel lifetime seemed to vary among the agonists. Channel conductance, which was independent of membrane potential, also varied significantly among the agonists. The average charge traversing the membrane through each open channel, calculated from the product of average channel lifetime and current, did not correlate with potency. Therefore, the dominant component of potency is the frequency of channel opening. No clear relationship between the structure of the agonist and channel lifetime or conductance was evident.


Effects of a propionic acid-based preservative on storage characteristics, nutritive value, and energy content for alfalfa hays packaged in large round bales.
During 2009 and 2010, alfalfa (Medicago sativa L.) hays from 2 cuttings harvested from the same field site were used to evaluate the effects of a propionic acid-based preservative on the storage characteristics and nutritive value of hays stored as large round bales. A total of 87 large round bales (diameter = 1.5m) were included in the study; of these, 45 bales served as controls, whereas 42 were treated with a commercial propionic acid-based preservative at mean application rates of 0.5±0.14 and 0.7±0.19% of bale weight, expressed on a wet (as is) or dry matter basis, respectively. Initial bale moisture concentrations ranged from 10.2 to 40.4%. Internal bale temperatures were monitored daily during an outdoor storage period, and heating characteristics were summarized for each bale as heating degree days (HDD) >30°C. For acid-treated bales, the regression relationship between HDD and initial bale moisture was best fitted to a quadratic model in which the linear term was dropped to improve fit (Y=2.02x(2) - 401; R(2)=0.77); control hays were best fitted to a nonlinear model in which the independent variable was squared [Y=4,112 - (4,549×e(-0.000559x*x)); R(2)=0.77]. Based on these regressions, acid-treated bales accumulated more HDD than control hays when the initial bale moisture was >27.7%; this occurred largely because acid treatment tended to prolong active heating relative to control hays. Linear regressions of recoveries of dry matter on HDD did not differ on the basis of treatment, yielding a common linear relationship of Y=-0.0066x+96.3 (R(2)=0.75). Regressions relating changes (post-storage - pre-storage) in concentrations of several nutritional components (neutral detergent fiber, lignin, ash, crude protein, and total digestible nutrients) with HDD for acid-treated hays typically exhibited more inflection points or were higher-ordered polynomial regressions than those of control hays. These more complex responses probably reflected the perturbation of normal heating patterns following acid treatment; however, overall effects on post-storage nutritive value were relatively limited in scope. The potential to improve nutritive value relative to cost for these large round bales was not especially favorable, and hay producers may find that diligence to achieve adequate field desiccation before baling, or use of oxygen-exclusion methods, such as wrapping in plastic, may be better alternatives for preserving moist hays.


The alteration of mitotic events by ionophore A23187 and carbonyl cyanide n-chlorophenylhydrazone.
A large quantity of published work indicates that calcium ions may be involved in the regulation of mitotic events and recent reports suggest that the onset of chromosome movement is dependent upon a transient increase in free cytosolic calcium ions. In this paper we examine the effects of two agents known to perturb intracellular calcium pools on mitosis in HeLa cells. These were the calcium-selective ionophore A23187 and carbonyl cyanide n-chlorophenylhydrazone (CCCP), which is a protonophoric inhibitor of oxidative phosphorylation. Owing to a stimulation of glycolysis, the latter agent does not decrease intracellular ATP in HeLa but does cause mitochondria to release calcium ions. Our data show that, at low concentrations, both agents prolong metaphase but differ in their effects on anaphase and cytokinesis. Studies with chlorotetracycline, a commonly used probe for membrane-associated calcium, verify that these agents do affect calcium pools under the conditions of our experiments. The data presented are consistent with the idea that increased cytosolic calcium levels can directly or indirectly affect mitotic events but, contrary to other suggestions, cause a prolongation of metaphase, i.e. they delay the onset of chromosome movement.


Childhood experiences and current adjustment of offspring of indigent patients with schizophrenia.
This study reports the childhood experiences, current life situation and level of adjustment, and prior mental health service use of offspring of indigent people with schizophrenia. Sixty-eight patient-parents were asked for consent for researchers to contact their adolescent and adult offspring. Thirty-nine consenting offspring were interviewed with an assessment battery that included measures of current occupational and social functioning, psychiatric status, and mental health service use. Interviewed offspring were raised in an average of three different settings from birth to 18 years of age. Relatives, particularly grandparents and aunts, were more likely to provide surrogate parenting than were nonkin foster parents and were more significant nurturing figures than biological parents. The typical offspring had a high school diploma, was gainfully employed, and was involved with a spouse or household partner or had a close friend. Twenty-three of the 39 offspring had children, and most were raising their children alone. Ten offspring had a diagnosis of major depression, schizoaffective disorder, or drug or alcohol abuse, but none had a diagnosis of schizophrenia. Four of the ten offspring with a psychiatric diagnosis had never been treated. Findings underscore the need for long-term studies of families with a parent who is a psychiatric patient. Rehabilitation efforts should include extended family who play a critical role in raising offspring during periods when patient-parents are unable to do so. Offspring should be included in efforts to educate families about schizophrenia.


Fertile green: green facilitates creative performance.
The present research sought to extend the nascent literature on color and psychological functioning by examining whether perception of the color green facilitates creativity. In four experiments, we demonstrated that a brief glimpse of green prior to a creativity task enhances creative performance. This green effect was observed using both achromatic (white, gray) and chromatic (red, blue) contrast colors that were carefully matched on nonhue properties, and using both picture-based and word-based assessments of creativity. Participants were not aware of the purpose of the experiment, and null effects were obtained on participants' self-reported mood and positive activation. These findings indicate that green has implications beyond aesthetics and suggest the need for sustained empirical work on the functional meaning of green.


Non-muscle-invasive Bladder Cancer: Side-by-Side Guideline Comparison.
Clinical guidelines from the European Association of Urology, American Urological Association, Society of Urologic Oncology, and National Comprehensive Cancer Network are some of the most frequently accessed publications. These guidelines are published at varying frequency and use different methods to formulate their recommendations. Many guidelines still rely on expert opinion in areas where there is a lack of data. To be well executed guidelines they need to involve comprehensive panels who are content experts and multispecialty. This article reviews the strengths and weaknesses of current guidelines for non-muscle-invasive bladder cancer and possible opportunities for future improvements. PATIENT SUMMARY: Quality recommendations in guidelines are critical to provide the most effective care for patients with non-muscle-invasive bladder cancer.


High LC3/Beclin Expression Correlates with Poor Survival in Glioma: a Definitive Role for Autophagy as Evidenced by In Vitro Autophagic Flux.
Recent studies suggest the role of autophagy, an evolutionarily conserved catabolic process, in determining the response of gliomas to treatment either positively or negatively. The study attempts to characterize autophagy in low and high-grade glioma by investigating the autophagic flux and clinical significance of autophagy proteins (LC3 and beclin 1) in a group of glioma patients. We evaluated the expression of autophagic markers in resected specimens of low-grade glioma (LGG) and high-grade glioma (HGG) tissues, by immunohistochemistry and Western blotting. Our results show that expression of autophagy proteins were more prominent in HGG than in LGG. Increased level of autophagic proteins in HGG can be due to an increased rate of autophagy or can be because of blockage in the final degradation step of autophagy (defective autophagy). To distinguish these possibilities, the autophagic flux assay which helps to determine the rate of degradation/synthesis of autophagic proteins (LC3-II and p62) over a period of time by blocking the final degradation step of autophagy using bafilomycin A1 was used . The assessment of autophagic flux in ex vivo culture of primary glioma cells revealed for the first time increased turnover of autophagy in high grade compared to low grade-glioma. Though autophagic markers were reduced in LGG, functionally autophagy was non defective in both grades of glioma. We then investigated whether autophagy in gliomas is regulated by nutrient sensing pathways including mTOR and promote cell survival by providing an alternate energy source in response to metabolic stress. The results depicted that the role of autophagy during stress varies with tissue and has a negative correlation with mTOR substrate phosphorylation. We also evaluated the expression of LC3 and beclin 1 with progression free survival (PFS) using Kaplan-Meier survival analysis and have found that patients with low LC3/beclin 1 expression had better PFS than those with high expression of LC3/beclin 1 in their tumors. Together, we provide evidence that autophagy is non-defective in glioma and also show that high LC3/beclin 1 expression correlates with poor PFS in both LGG and HGG.


The impact of object and gesture imitation training on language use in children with autism spectrum disorder.
Reciprocal imitation training (RIT) is a naturalistic behavioral intervention that teaches imitation to children with autism spectrum disorder (ASD) within a social-communicative context. RIT has been shown to be effective at teaching spontaneous, generalized object and gesture imitation. In addition, improvements in imitation are associated with increases in verbal imitation and spontaneous language. This study used a modified multiple-baseline design across 4 children to examine whether adding gesture imitation training improves the overall rate of appropriate language use in children with ASD who have already been participating in object imitation training. Three of the 4 children showed greater improvements in their use of appropriate language after gesture imitation was begun. Further, the children were more likely to use verbal imitation during gesture imitation training than during object imitation training. These findings suggest that adding gesture imitation training to object imitation training can lead to greater gains in rate of language use than object imitation alone. Implications for both language development and early intervention are discussed.


How a cell deals with abnormal proteins. Pathogenetic mechanisms in protein aggregation diseases.
Defective protein folding is responsible for many diseases. Although these diseases seem to be quite diverse at the first glance, there is evidence for common pathogenetic principles. The basis of the pathological changes is the cell's inability to prevent protein misfolding, to revert misfolded proteins to normal or to eliminate misfolded proteins by degradation. This could result in deposition of potentially cytotoxic protein aggregates (protein aggregation diseases). Chronic degenerative diseases of the central nervous system (e.g. Alzheimer's and Parkinson's disease), the amyloidoses, but also chronic liver diseases, for example alcoholic and nonalcoholic steatohepatitis, belong to this category of disorders. This review highlights general pathogenic principles of protein aggregation diseases based on immunohistochemical and biochemical studies as well as observations in a mouse model for protein aggregation in the context of alcoholic and nonalcoholic steatohepatitis. The cellular defense mechanisms involved in protein quality control as well as the pathogenesis of protein aggregation diseases will be discussed.


Effects of drugs on mucus clearance.
Mucociliary clearance (MCC), the process in which airway mucus together with substances trapped within are moved out of the lungs, is an important defence mechanism of the human body. Drugs may alter this process, such that it is necessary to know the effect of the drugs on MCC. Indeed, agents stimulating MCC may be used therapeutically in respiratory medicine, especially in patients suspected of having an impairment of their mucociliary transport system. In contrast, caution should be taken with drugs depressing MCC as an undesired side-effect, independently of their therapeutic indication. Since cough clearance (CC) serves as a back-up system when MCC fails, the influence of drugs must be examined not only on MCC but also on CC. Ultimately, the clinical repercussions of alterations in mucus transport induced by drug administration must be studied. Tertiary ammonium compounds (anticholinergics), aspirin, anaesthetic agents and benzodiazepines have been shown to be capable of depressing the mucociliary transport system. Cholinergics, methylxanthines, sodium cromoglycate, hypertonic saline, saline as well as water aerosol have been shown to increase MCC. Adrenergic antagonists, guaifenesin, S-carboxymethylcysteine, sodium 2-mercapto-ethane sulphonate and frusemide have been reported not to alter the mucociliary transport significantly. Amiloride, uridine 5'-triphosphate (UTP), quaternary ammonium compounds (anticholinergics), adrenergic agonists, corticosteroids, recombinant human deoxyribonuclease (rhDNase), N-acetylcysteine, bromhexine and ambroxol have been reported either not to change or to augment MCC. Indirect data suggest that surfactant as well as antibiotics may improve the mucociliary transport system. As for the influence of drugs on CC, amiloride and rhDNase have been demonstrated to increase the effectiveness of cough. A trend towards an improved CC was noted after treatment with adrenergic agonists. The anticholinergic agent ipratropium bromide, which is a quaternary ammonium compound, has been suggested to decrease CC significantly. Bromhexine, ambroxol and neutral saline seemed not to alter CC, either positively or negatively. Finally, treatment with either amiloride, recombinant human deoxyribonuclease, bromhexine, ambroxol, N-acetylcysteine, S-carboxymethylcysteine or hypertonic saline has been suggested as a possible cause of clinical improvement in patients, such as the experience of dyspnoea, the case of expectoration or the frequency of infective exacerbations. Other agents did not show a clinical benefit.


Incomplete revascularization for percutaneous coronary interventions: Variation among operators, and association with operator and hospital characteristics.
Many studies have compared outcomes for incomplete revascularization (IR) among patients undergoing percutaneous coronary interventions (PCIs), but little is known about the correlates of IR, the extent to which complete revascularization (CR) was attempted unsuccessfully, and the variation across operators in the use of IR. New York's PCI registry was used to examine medium-term mortality for IR, the variables associated with the use of IR, and the variation across operators in the utilization of IR after controlling for patient factors. Incomplete revascularization occurred for 63% of all patients and was significantly associated with higher 3-year mortality (adjusted hazard ratio1.35, 95% CI 1.23-1.48) than for CR. A total of 96% of all attempted CRs were successful. Operators with 15 or fewer years in practice (the lowest half) used IR significantly more (65% vs 61%, adjusted odds ratio [AOR] 1.17, 95% CI 1.00-1.37) than other operators, and operators with annual volumes of 171 or lower (the lowest 3 quartiles) used IR more than other operators (68% vs 60%, AOR 1.35, 95% CI 1.14-1.59). Also, hospitals with annual volumes of 645 and lower (the lowest 50% of hospitals) used IR more (67% vs 62%, AOR 1.46, 95% CI 1.07-1.99) than other hospitals. Percutaneous coronary intervention patients without myocardial infarction who undergo IR continue to have higher medium-term (3-year) risk-adjusted mortality rates. There is a large amount of variability among operators in the frequency with which IR occurs. Operators who have been in practice longer, and higher-volume operators and hospitals have lower rates of IR. Failed attempts at CR occur very infrequently.


Food allergy knowledge of parents - is ignorance bliss?
Food allergic children are at least partially dependent on their parents to care for their food allergy. In addition, parents are often responsible for the education of others regarding food allergy, including the family, school, neighbors, and friends. The aim of this study was to investigate food allergy knowledge, attitudes, and beliefs of parents with food allergic children in the Netherlands. In addition, a cross-cultural comparison was made between parents from the USA and parents from the Netherlands. The original Chicago Food Allergy Research Survey for Parents of Children with Food Allergy (CFARS-PRNT) was translated into Dutch. Parents of children with at least one doctor-diagnosed food allergy were included. Knowledge scores and attitude/beliefs scores were determined and compared with the data from 2945 parents from the USA. Predictors of overall knowledge scores were investigated. Dutch parents of children completed the translated CFARS-PRNT (n = 299). The mean overall knowledge score in the Netherlands was 9.9 after adjusting for guessing, compared with 12.7 in the USA (p < 0.001). Attitudes and beliefs regarding food allergy among parents from the Netherlands were generally more optimistic. The overall knowledge scores could be predicted by country of origin, educational degree, being member of a patient organization, visiting an allergist, and a history of anaphylaxis. Food allergy knowledge among parents of food allergic children from the Netherlands is suboptimal when compared with their counterparts from the USA, although these parents tend to be more optimistic toward food allergy than parents from the USA.


[The immunological phenotyping of the lymphocytes in HIV+ patients].
We propose that the ratios of lymphocyte subsets: CD4+/CD8+, CD4+/CD3+, and CD8+/CD3+, graphically displayed in this order, could be an easy way of immunophenotyping and a simple form of interpretation both for clinicians and patients. A total of 187 asymptomatic HIV-positive patients, -including 83 symptomatic patients with AIDS- and a normal comparative age group of 36 patients, were studied by flow cytometry. Ratios were graphically displayed on millimetric-grid paper, and a specific point was given to each obtained ratio. The points were connected by a line. The angles formed by the lines were measured and graphically displayed over a circle. The point of reference 0 degree was located on the left side of the circle, giving the corresponding 90 degrees, 180 degrees, and 240 degrees angles. The graph of the control group shows a "beach chair" image. As soon as the CD4+ lymphocytes diminish and the CD8+ lymphocytes increase in HIV+ patients, "the chair's headrest" descends and the "chair's footpiece" swivels back. Given the ranks and fluctuations of the lymphocyte subsets, their graphic display provides a better representation of the immunological status.


Evaluation of optical changes of three types of lenses: hard PMMA, hydrogel, and heparin surface modified hard lenses effected by silicone oil, used clinically as a substitute of the vitreous body.
The objective of these investigations was an in vitro evaluation whether silicone oil OXANE of viscosity 5700 cSt clinically used in eye surgery as a substitute of the vitreous body, being in contact with an artificial polymer lens used as an implant of human lens, causes the changes in its optical properties. The paper presents the results of spectral analysis of transmission of visible (VIS) radiation of three types of artificial lenses: hard PMMA, hydrogel, heparin surface modified (HSM) hard PMMA, and the same lenses damaged by YAG laser radiation with an energy increasing from 1.7 mJ to 3.7 mJ, exposed to clinically applied silicone oil. The studies were carried out, in two-week intervals, over a period of 20 weeks. Hard PMMA and HSM lenses were found not to have changed their optical properties after 20 weeks of exposure to silicone oil. The measured transmittance values were within the range of instrumental error (+/- 1%). Optical properties of hydrogel lenses exposed to silicone oils deteriorated with exposure and after 20-week exposure to silicone oil the average transmittance value decreased by about 18%, reaching its final value of 67.08 +/- 2.37% (RSD = 5.56%). A minimal decrease of the initial transmittance values was observed only for the lenses exposed to laser radiation of highest energy (3.7 mJ). After completed exposure to silicone oil, two kinds of lenses were found to have a slightly improved transmittance: hard PMMA lenses by about 4% and HSM lenses by about 2%. On the other hand, in case of hydrogel lenses the deterioration of optical properties to the extent comparable to that of hydrogel lenses not damaged by laser radiation was observed.


Epigenetic abnormality of SRY gene in the adult XY female with pericentric inversion of the Y chromosome.
In normal ontogenetic development, the expression of the sex-determining region of the Y chromosome (SRY) gene, involved in the first step of male sex differentiation, is spatiotemporally regulated in an elaborate fashion. SRY is expressed in germ cells and Sertoli cells in adult testes. However, only few reports have focused on the expressions of SRY and the other sex-determining genes in both the classical organ developing through these genes (gonad) and the peripheral tissue (skin) of adult XY females. In this study, we examined the gonadal tissue and fibroblasts of a 17-year-old woman suspected of having disorders of sexual differentiation by cytogenetic, histological, and molecular analyses. The patient was found to have the 46,X,inv(Y)(p11.2q11.2) karyotype and streak gonads with abnormally prolonged SRY expression. The sex-determining gene expressions in the patient-derived fibroblasts were significantly changed relative to those from a normal male. Further, the acetylated histone H3 levels in the SRY region were significantly high relative to those of the normal male. As SRY is epistatic in the sex-determination pathway, the prolonged SRY expression possibly induced a destabilizing effect on the expressions of the downstream sex-determining genes. Collectively, alterations in the sex-determining gene expressions persisted in association with disorders of sexual differentiation not only in the streak gonads but also in the skin of the patient. The findings suggest that correct regulation of SRY expression is crucial for normal male sex differentiation, even if SRY is translated normally.


Heat shock protein 47 promotes tumor survival and therapy resistance by modulating AKT signaling via PHLPP1 in colorectal cancer.
Objective: Heat shock protein 47 (HSP47) is a collagen-specific molecular chaperone that facilitates collagen maturation. Its role in cancer remains largely unknown. In this study, we investigated the roles of HSP47 in colorectal cancer (CRC) and therapy resistance. Methods: Expression of HSP47 in CRC tissues was examined (1) in paired human CRC/adjacent normal tissues, using real time quantitative reverse transcription polymerase chain reaction (qRT-PCR), The Cancer Genome Atlas (TCGA) database, and 22 independent microarray databases (curated CRC). In vitro studies on several CRC cell lines (HCT116, RKO and CCL228) with modulated HSP47 expression were conducted to assess cell viability and apoptosis (TUNEL assay and caspase-3/-7) during exposure to chemotherapy. AKT signaling and co-immunoprecipitation studies were performed to examine HSP47 and PHLPP1 interaction. In vivo studies using tumor xenografts were conducted to assess the effects of HSP47 modulation on tumor growth and therapy response. Results: HSP47 was upregulated in CRC and was associated with poor prognosis in individuals with CRC. In vitro, HSP47 overexpression supported the survival of CRC cells, whereas its knockdown sensitized cells to 5-fluorouracil (5-FU). HSP47 promoted survival by inhibiting apoptosis, enhancing AKT phosphorylation, and decreasing expression of the AKT-specific phosphatase PHLPP1 when cells were exposed to chemotherapy. These effects were partly results of the interaction between HSP47 and PHLPP1, which decreased PHLPP1 stability and led to more persistent AKT activity. In vivo, HSP47 supported tumor growth despite 5-FU treatment. Conclusions: HSP47 supports the growth of CRC tumors and suppresses the efficacy of chemotherapy via modulation of AKT signaling.


Sera selected from national STI surveillance system shows Chlamydia trachomatis PgP3 antibody correlates with time since infection and number of previous infections.
Seroprevalence surveys of Chlamydia trachomatis (CT) antibodies are promising for estimating age-specific CT cumulative incidence, however accurate estimates require improved understanding of antibody response to CT infection. We used GUMCAD, England's national sexually transmitted infection (STI) surveillance system, to select sera taken from female STI clinic attendees on the day of or after a chlamydia diagnosis. Serum specimens were collected from laboratories and tested anonymously on an indirect and a double-antigen ELISA, both of which are based on the CT-specific Pgp3 antigen. We used cross-sectional and longitudinal descriptive analyses to explore the relationship between seropositivity and a) cumulative number of chlamydia diagnoses and b) time since most recent chlamydia diagnosis. 919 samples were obtained from visits when chlamydia was diagnosed and 812 during subsequent follow-up visits. Pgp3 seropositivity using the indirect ELISA increased from 57.1% (95% confidence interval: 53.2-60.7) on the day of a first-recorded chlamydia diagnosis to 89.6% (95%CI: 79.3-95.0) on the day of a third or higher documented diagnosis. With the double-antigen ELISA, the increase was from 61.1% (95%CI: 53.2-60.7) to 97.0% (95%CI: 88.5-99.3). Seropositivity decreased with time since CT diagnosis on only the indirect assay, to 49.3% (95%CI: 40.9-57.7) two or more years after a first diagnosis and 51.9% (95%CI: 33.2-70.0) after a repeat diagnosis. Seropositivity increased with cumulative number of infections, and decreased over time after diagnosis on the indirect ELISA, but not on the double-antigen ELISA. This is the first study to demonstrate the combined impact of number of chlamydia diagnoses, time since diagnosis, and specific ELISA on Pgp3 seropositivity. Our findings are being used to inform models estimating age-specific chlamydia incidence over time using serial population-representative serum sample collections, to enable accurate public health monitoring of chlamydia.


Training and academic demands are associated with sleep quality in high-performance "dual career" student swimmers.
Dual career collegiate athletes, who combine sporting endeavours with academic studies, are commonplace in high-performance sport. Sleep is an important aspect of physical and psychological recovery for athletes, plays a role in memory and learning and is associated with academic achievement. The aim of this study was to assess variations in sleep characteristics of collegiate swimmers to understand the incidence of poor sleep, and which stressors might contribute towards this. A total of 22 (male n = 13, female n = 9) high-performance student swimmers (mean ± SD; age 20 ± 2 years) participated in this study. Sleep/wake behaviour was assessed using the Pittsburgh Sleep Quality Index each month over a 12-month period. Additionally, academic and sporting commitments were quantified on a monthly basis and summarised as 4 stressors: average weekly training hours, total number of competitive races, total number of academic assessments, and average weekly learning hours. On average across the 12-month period, 41.7% of athletes reported poor sleep. The latest bedtimes, wake times and longest sleep durations were found in the months where academic and training demands were lowest. A statistically significant positive association was identified between median sleep quality scores and mean number of academic assessments (ρ(12) = 0.71, p=0.005). Hierarchal linear modelling analysis determined that number of academic assessments best predicted sleep quality within this cohort. The present study highlights the need for those working with athletes who have competing academic demands to consider sleep quality, and its potential impact on performance and wellbeing.


Specific association of Piwi with rasiRNAs derived from retrotransposon and heterochromatic regions in the Drosophila genome.
In Drosophila, Piwi (P-element-induced wimpy testis), which encodes a protein of the Argonaute family, is essential for germ stem cell self-renewal. Piwi has recently been shown to be a nuclear protein involved in gene silencing of retrotransposons and controlling their mobilization in the male germline. However, little is known about the molecular mechanisms of Piwi-dependent gene silencing. Here we show that endogenous Piwi immunopurified from ovary specifically associates with small RNAs of 25-29 nucleotides in length. Piwi-associated small RNAs were identified by cloning and sequencing as repeat-associated small interfering RNAs (rasiRNAs) derived from repetitive regions, such as retrotransposon and heterochromatic regions, in the Drosophila genome. Northern blot analyses revealed that in vivo Piwi does not associate with microRNAs (miRNAs) and that guide siRNA was not loaded onto Piwi when siRNA duplex was added to ovary lysate. In vitro, recombinant Piwi exhibits target RNA cleavage activity. These data together imply that Piwi functions in nuclear RNA silencing as Slicer by associating specifically with rasiRNAs originating from repetitive targets.


Insights into the skin microbiome of sickle cell disease leg ulcers.
Leg ulcers are estimated to occur in 1%-10% of North American patients with sickle cell disease (SCD). Their pathophysiology remains poorly defined, but as with other chronic wounds, it is hypothesised that the microbial milieu, or microbiome, contributes to their healing and clinical outcomes. This study utilises 16S ribosomal RNA (rRNA) gene sequencing to describe, for the first time, the microbiome of the SCD leg ulcer and its association with clinical factors. In a cross-sectional analysis of 42 ulcers, we recovered microbial profiles similar to other chronic wounds in the predominance of anaerobic bacteria and opportunistic pathogens including Staphylococcus, Corynebacterium, and Finegoldia. Ulcers separated into two clusters: one defined by predominance of Staphylococcus and smaller surface area, and the other displaying a greater diversity of taxa and larger surface area. We also find that the relative abundance of Porphyromonas is negatively associated with haemoglobin levels, a key clinical severity indicator for SCD, and that Finegoldia relative abundance is negatively associated with CD19+ B cell count. Finally, ratios of Corynebacterium:Lactobacillus and Staphylococcus:Lactobacillus are elevated in the intact skin of individuals with a history of SCD leg ulcers, while the ratio of Lactobacillus:Bacillus is elevated in that of individuals without a history of ulcers. Investigations of the skin microbiome in relation to SCD ulcer pathophysiology can inform clinical guidelines for this poorly understood chronic wound, as well as enhance broader understanding about the role of the skin microbiome in delayed wound healing.


Influence of x-ray diffraction sample preparation on quantitative mineralogy: implications for chromate waste treatment.
Powders of chromite ore processing residue (COPR) were mineralogically evaluated using quantitative X-ray powder diffraction (XRPD) to illustrate the impacts of sample preparation procedures. Chromite ore processing residue is strongly alkaline, reactive, contains minerals of varying hardness and absorption coefficients, and exhibits significant amorphicity. This poses a challenge to produce powders for XRPD analysis that are sufficiently fine and of uniform particle size while avoiding mineral reactions and overgrinding effects. Dry, hand pulverization to different grain sizes, and wet, mechanical pulverization (micromilling) using four milling liquids (cyclohexane, isopropanol, ethanol, and water), and variable milling durations (up to 15 min) were evaluated. Micromilling with a light, nonpolar, highly evaporative liquid such as cyclohexane with a milling time of 5 min mitigated systematic errors such as microabsorption and preferred orientation as it produced finer and more uniform particle size distributions than the hand-pulverized powders, while simultaneously affording the least time for sample preparation. Conversely, the use of water as milling liquid resulted in extensive hydration reactions during sample preparation, causing mischaracterization and significant underestimation of its reactive brownmillerite content, which can complicate the remediation design process for COPR. Hand pulverization emerged as a necessary complement to quantify Cr(VI)-containing, softer minerals destroyed during mechanical milling, the quantification of which has also important implications for COPR treatment design. The findings of this study may be applicable in a variety of geochemically complicated and reactive environmental media (metal-contaminated soils, stabilized/solidified media, inorganic waste), and points to the importance of the sample preparation method to obtain reliable quantitative XRPD results.


The interaction of Acanthamoeba castellanii cysts with macrophages and neutrophils.
Acanthamoeba castellanii, a free-living amoeba, causes a sight-threatening form of keratitis. Even after extensive therapies, corneal damage can be severe, often requiring corneal transplantation to restore vision. However, A. castellanii cysts are not eliminated from the conjunctiva and stroma of humans and can excyst, resulting in infection of the corneal transplant. The aim of this study was to determine whether elements of the innate immune apparatus, neutrophils and macrophages, were capable of detecting and eliminating A. castellanii cysts and to examine the mechanism by which they kill the cysts. Results show that neither innate immune cell is attracted chemotactically to intact cysts, yet both were attracted to lysed cysts. Both macrophages and neutrophils were capable of killing significant numbers of cysts, yet neutrophils were 3-fold more efficient than macrophages. Activation of macrophages with lipopolysaccharide and interferon-gamma did not increase their cytolytic ability. Conditioned medium isolated from macrophages did not lyse the cysts; however, prevention of phagocytosis by cytochalasin D inhibited 100% of macrophage-mediated killing of the cysts. Conditioned medium from neutrophils did kill significant numbers of the cysts, and this killing was blocked by quercetin, a potent inhibitor of myeloperoxidase (MPO). These results indicate that neither macrophages nor neutrophils are chemoattracted to intact cysts, yet both are capable of killing the cysts. Macrophages killed the cysts by phagocytosis, whereas neutrophils killed cysts through the secretion of MPO.


Adding Value to the Health Care System: Identifying Value-Added Systems Roles for Medical Students.
To catalyze learning in Health Systems Science and add value to health systems, education programs are seeking to incorporate students into systems roles, which are not well described. The authors sought to identify authentic roles for students within a range of clinical sites and explore site leaders' perceptions of the value of students performing these roles. From 2013 to 2015, site visits and interviews with leadership from an array of clinical sites (n = 30) were conducted. Thematic analysis was used to identify tasks and benefits of integrating students into interprofessional care teams. Types of systems roles included direct patient benefit activities, including monitoring patient progress with care plans and facilitating access to resources, and clinic benefit activities, including facilitating coordination and improving clinical processes. Perceived benefits included improved value of the clinical mission and enhanced student education. These results elucidate a framework for student roles that enhance learning and add value to health systems.


Autodigestion: Proteolytic Degradation and Multiple Organ Failure in Shock.
There is currently no effective treatment for multiorgan failure following shock other than supportive care. A better understanding of the pathogenesis of these sequelae to shock is required. The intestine plays a central role in multiorgan failure. It was previously suggested that bacteria and their toxins are responsible for the organ failure seen in circulatory shock, but clinical trials in septic patients have not confirmed this hypothesis. Instead, we review here evidence that the digestive enzymes, synthesized in the pancreas and discharged into the small intestine as requirement for normal digestion, may play a role in multiorgan failure. These powerful enzymes are nonspecific, highly concentrated, and fully activated in the lumen of the intestine. During normal digestion they are compartmentalized in the lumen of the intestine by the mucosal epithelial barrier. However, if this barrier becomes permeable, e.g. in an ischemic state, the digestive enzymes escape into the wall of the intestine. They digest tissues in the mucosa and generate small molecular weight cytotoxic fragments such as unbound free fatty acids. Digestive enzymes may also escape into the systemic circulation and activate other degrading proteases. These proteases have the ability to clip the ectodomain of surface receptors and compromise their function, for example cleaving the insulin receptor causing insulin resistance. The combination of digestive enzymes and cytotoxic fragments leaking into the central circulation causes cell and organ dysfunction, and ultimately may lead to complete organ failure and death. We summarize current evidence suggesting that enteral blockade of digestive enzymes inside the lumen of the intestine may serve to reduce acute cell and organ damage and improve survival in experimental shock.


Bioinformatics and in-silico findings reveal medical features and pharmacological targets of biochanin A against colorectal cancer and COVID-19.
Severe mortality due to the COVID-19 pandemic resulted from the lack of effective treatment. Although COVID-19 vaccines are available, their side effects have become a challenge for clinical use in patients with chronic diseases, especially cancer patients. In the current report, we applied network pharmacology and systematic bioinformatics to explore the use of biochanin A in patients with colorectal cancer (CRC) and COVID-19 infection. Using the network pharmacology approach, we identified two clusters of genes involved in immune response (IL1A, IL2, and IL6R) and cell proliferation (CCND1, PPARG, and EGFR) mediated by biochanin A in CRC/COVID-19 condition. The functional analysis of these two gene clusters further illustrated the effects of biochanin A on interleukin-6 production and cytokine-cytokine receptor interaction in CRC/COVID-19 pathology. In addition, pathway analysis demonstrated the control of PI3K-Akt and JAK-STAT signaling pathways by biochanin A in the treatment of CRC/COVID-19. The findings of this study provide a therapeutic option for combination therapy against COVID-19 infection in CRC patients.


Handheld computers for data entry: high tech has its problems too.
The use of handheld computers in medicine has increased in the last decade, they are now used in a variety of clinical settings. There is an underlying assumption that electronic data capture is more accurate that paper-based data methods have been rarely tested. This report documents a study to compare the accuracy of hand held computer data capture versus more traditional paper-based methods. Clinical nurses involved in a randomised controlled trial collected patient information on a hand held computer in parallel with a paper-based data form. Both sets of data were entered into an access database and the hand held computer data compared to the paper-based data for discrepancies. Error rates from the handheld computers were 67.5 error per 1000 fields, compared to the accepted error rate of 10 per 10,000 field for paper-based double data entry. Error rates were highest in field containing a default value. While popular with staff, unacceptable high error rates occurred with hand held computers. Training and ongoing monitoring are needed if hand held computers are to be used for clinical data collection.


Pterostilbene (3',5'-dimethoxy-resveratrol) exerts potent antitumor effects in HeLa human cervical cancer cells via disruption of mitochondrial membrane potential, apoptosis induction and targeting m-TOR/PI3K/Akt signalling pathway.
To examine the molecular mechanism of action behind the anticancer effects of pterostilbene in HeLa human cervical cancer cells. MTS assay was used to study the pterostilbene cytotoxic effects, while inverted phase contrast and fluorescence microscopy was used to study the effects of the drug on the cell apoptosis and changes in cell morphology. Flow cytometry was used to study changes in mitochondrial membrane potential (MMP), while immunoblotting assay was used to demonstrate its effects on m-TOR/PI3K/Akt protein signalling pathway. Pterostilbene induced potent, dose-dependent and time-dependent cytotoxic effects in HeLa cancer cells exhibiting IC50 of 101.2 µM and 65.9 µM at 24 and 48 hrs time intervals, respectively. As compared to the untreated control cells which revealed normal cell morphology, pterostilbene-treated cells exhibited significant cellular shrinkage which increased with increasing doses of the drug. Untreated control cells showed complete green fluorescence corresponding to absence of apoptosis. However, pterostilbene-treated cells with 25, 100 and 200 µM showed increasing emission of red/orange fluorescence corresponding to apoptotic induction. Pterostilbene-treated cells showed evident signs of DNA ladder formation and the effect increased with increasing concentrations of the drug. The percentage of cells with depolarized mitochondria (loss of MMP) increased from 2.3% in the control group to 24.5, 43.2 and 65.8% in cells treated with 0, 25, 100 and 200 µM concentration of pterostilbene, respectively. Pterostilbene exerts potent anticancer effects in HeLa human cervical cancer cells via disruption of MMP, apoptosis induction and targeting m-TOR/PI3K/Akt signalling pathway.


[Transient prosopagnosia and lasting topographical disorientation after the total removal of a right occipital arteriovenous malformation].
A 32-year-old right-handed female tour guide developed a transient prosopagnosia after the total removal of an arteriovenous malformation in the right occipital lobe. Neurological examination revealed only a left homonymous hemianopsia. The ablation on the right side involved total occipital lobe, the posterior region of the parahippocampal gyrus, and a part of the precuneus and angular gyrus. Neuropsychologically she was alert and oriented in time and place. No aphasia or apraxia was observed. Visual recognition of objects, colors and letters was normal. She could match faces and insects easily, despite a close resemblance among them. Her prosopagnosia resolved in one month, while her impairment in topographical orientation and nonverbal memory continued more than eight months. These findings exclude the possibility that her prosopagnosia was derived from visual agnosia, poor discrimination of details, or nonverbal memory deficit. Excluding cases with malignant brain tumor which may affect more diffuse area of the brain, there was only one reported case who developed persistent prosopagnosia after the right occipital lobectomy. Given the rarity of prosopagnosics with only right-sided lesions, it seems that in most of humans posterior parts of both hemispheres participate in the identification of faces.


[Endometrial cancer in young patients: report of 17 cases].
Endometrial cancer of young (less then 40 years old) patients comprises 4-5% of all endometrial cancers in Hungary. The majority of patients did not give birth yet, so fertility sparing is very important. Fertility sparing treatment is possible if the tumor's histology is endometrial type and Grade 1 (well differentiated). The tumor localizes only to the endometrium and there is no myometrium infiltration. The authors present 17 cases of patients treated at the Department of Gynecology of National Institute of Oncology (Budapest, Hungary). In 3 cases conservative therapy (progesterone treatment) was possible, and 14 patients had to undergo surgery, because conservative treatment did fail. Of 17 patients 14 were never pregnant. The average patient's age was 32.35 ± 4.27 years. The mean body weight was 93.13 ± 30.79 kg (from 58 kg up to 147 kg); in 7 cases BMI (body mass index) was more than 30. After surgery histological examination had revealed 2 cases with normal ovaries, 1 case of simple cyst and 1 case of malignant ovarian tumor (serous adenocarcinoma, Grade 2), and 10 cases of polycystic ovaries associated with endometrial cancer. Of 3 cases that had only curettage, the endometrial cancer was Grade 1, and in 1 case radiological imaging showed simplex ovarian cyst. The authors' findings concerning young endometrial cancer patients confirm the results published in the literature. In cases suitable for fertility sparing treatment it is not sufficient to concentrate only on endometrial findings, but is very important to focus on the therapy of cystic ovaries (80% of which is PCO), obesity and diabetes mellitus as well.


Statistical characterization of hyperspectral background clutter in the reflective spectral region.
Hyperspectral imaging systems for daylight operation measure and analyze reflected and scattered radiation in p-spectral channels covering the reflective infrared region 0.4-2.5 microm. Consequently, the p-dimensional joint distribution of background clutter is required to design and evaluate optimum hyperspectral imaging processors. In this paper, we develop statistical models for the spectral variability of natural hyperspectral backgrounds using the class of elliptically contoured distributions. We demonstrate, using data from the NASA AVIRIS sensor, that models based on the multivariate t-elliptically contoured distribution capture with sufficient accuracy the statistical characteristics of natural hyperspectral backgrounds that are relevant to target detection applications.


Ataxia Telangiectasia. A case report.
The case of an eight year old girl with Ataxia Telangiectasia (AT) is described. She presented at seven years of age with gait problems and was found to have the neurological, dermatological and immunological features characteristic of AT along with a history of frequent sino-pulmonary infections. This report highlights the refractory nature of the disease, the difficulties in medical management, and the problems posed by late diagnosis which can compromise patient care. This is a rare inherited form of ataxia which has not been previously reported in West Indian literature.


Obesity in America.
Recent data indicate that Americans are gaining weight at an alarming rate. In fact, data from the CDC indicate that the U.S. obesity average was 12% in 1990 and had grown to 23% by 2005. In recent years, this problem appears to be more prominent in some southern states than in other states. The purpose of this study was to determine what factors were associated with increased levels of obesity. Do demographic, educational, healthcare, or economic factors correlate with this trend? Using state level data in a fixed effects regression model we examined obesity rates for the period 1990-2003. We also used cross tabulation tables to compare obesity rates to several independent variables. Our analysis revealed that obesity was related to several health, demographic, and economic factors. As a result, we argue that policy makers as well as health officials should take a comprehensive look at obesity as well as other social ills, health care conditions, and related issues prior to creating a plan to improve health in this country.


Effects of Nicotinamide Riboside on Endocrine Pancreatic Function and Incretin Hormones in Nondiabetic Men With Obesity.
Augmenting nicotinamide adenine dinucleotide (NAD+) metabolism through dietary provision of NAD+ precursor vitamins translates to improved glucose handling in rodent models of obesity and diabetes. Preclinical evidence suggests that the NAD+/SIRT1 axis may be implicated in modulating important gut-related aspects of glucose regulation. We sought to test whether NAD+ precursor supplementation with nicotinamide riboside (NR) affects β-cell function, α-cell function, and incretin hormone secretion as well as circulating bile acid levels in humans. A 12-week randomized, double-blind, placebo-controlled, parallel-group trial in 40 males with obesity and insulin resistance allocated to NR at 1000 mg twice daily (n = 20) or placebo (n = 20). Two-hour 75-g oral glucose tolerance tests were performed before and after the intervention, and plasma concentrations of glucose, insulin, C-peptide, glucagon, glucagon-like peptide 1 (GLP-1), and glucose-dependent insulinotropic polypeptide (GIP) were determined. β-Cell function indices were calculated based on glucose, insulin, and C-peptide measurements. Fasting plasma concentrations of bile acids were determined. NR supplementation during 12 weeks did not affect fasting or postglucose challenge concentrations of glucose, insulin, C-peptide, glucagon, GLP-1, or GIP, and β-cell function did not respond to the intervention. Additionally, no changes in circulating adipsin or bile acids were observed following NR supplementation. The current study does not provide evidence to support that dietary supplementation with the NAD+ precursor NR serves to impact glucose tolerance, β-cell secretory capacity, α-cell function, and incretin hormone secretion in nondiabetic males with obesity. Moreover, bile acid levels in plasma did not change in response to NR supplementation.


Organically modified aluminosilicate mesostructures from block copolymer phases
Organically modified aluminosilicate mesostructures were synthesized from two metal alkoxides with the use of poly(isoprene-b-ethyleneoxide) block copolymers (PI-b-PEO) as the structure-directing molecules. By increasing the fraction of the inorganic precursors with respect to the polymer, morphologies expected from the phase diagrams of diblock copolymers were obtained. The length scale of the microstructures and the state of alignment were varied using concepts known from the study of block copolymers. These results suggest that the use of higher molecular weight block copolymer mesophases instead of conventional low-molecular weight surfactants may provide a simple, easily controlled pathway for the preparation of various silica-type mesostructures that extends the accessible length scale of these structures by about an order of magnitude.


Challenges in the Analysis of Gangliosides by LC-MS.
Gangliosides are a family of conjugates consisting of a polar sialoglycan head group and a hydrophobic ceramide tail. Gangliosides are of high abundance in neuronal tissues and are involved in numerous biological processes, such as cell-cell recognition, adhesion, and signal transduction. Alteration of the ganglioside profile is associated with various neurodegenerative diseases and there is indication that gangliosides are involved in the pathogenesis of Parkinson's and Huntington's disease. The development of refined methods for the analysis of gangliosides by high-performance liquid chromatography coupled to mass spectrometry (HPLC-MS) has supported research with qualitative and quantitative data. However, the amphiphilic character of gangliosides renders their separation and mass spectrometric analysis challenging. In this article, the strengths of hydrophilic interaction liquid chromatography (HILIC) for baseline separation of gangliosides, including two structural isomers, and their structural characterization by tandem mass spectrometry are demonstrated. The importance of ion source parameter optimization is highlighted to prevent misleading ganglioside transformation due to in-source dissociation.


Death by suicide during COVID-19 infection: Two case reports.
A range of psychiatric disorders has been recognized with coronavirus disease 2019 (COVID-19) infection, including acute stress, anxiety, depression, suicidal behavior, and post-traumatic stress disorder. Among those, the most worrying is death by suicide, which has been associated with COVID-19-related psychiatric disorders and psychosocial stressors. We report the first two cases of death by suicide, unlikely due to any current psychiatric disorders, while undergoing treatment in two inpatient facilities designated for COVID-19 patients. Case 1 was a 40-year-old man who presented to the emergency department with symptoms of a viral infection. This led to the diagnosis of COVID-19. While undergoing treatment in an inpatient facility, 3 weeks later, he died by hanging. Case 2 was a 25-year-old man with COVID-19-related upper respiratory tract symptoms and a possible undiagnosed pre-existing anxiety disorder. While undergoing treatment in a medical unit of a COVID-19-designated hospital, a week after the diagnosis of COVID-19, the patient died after jumping off the multistory hospital building. In both cases, there had been a diagnosis of COVID-19, and treatment was provided within an inpatient facility. Both patients were unvaccinated and had no evidence of a current psychiatric disorder or any warning signs of suicidal intent. Death by suicide can occur in COVID-19 patients without any warning signs of a psychiatric disorder or evidence of any apparent distress. Therefore, even without a diagnosable mental disorder, clinicians should still be vigilant about potential suicidal risk in patients with COVID-19 infection.


Effect of mycorrhizae, Thiobacillus and sulfur nutrition on the chemical composition of soybean [Glycine max (L.)] Merr. seed.
A field experiment carried out in a calcareous soil with a low available phosphorus to evaluate effectiveness of biofertilizers, mycorrhizae (Glomus intraradices) and Thiobacillus sp. inoculation individually or in combination on seed yield, oil, protein and some elements (P, Fe, Mn, Zn) concentration in two soybean [Glycine max (L.) Merr.] cultivars. The applied treatments were different fertilizers with 6 levels (including: NP (control, 12 kg N ha(-1) as urea, 46 kg P2O5 ha(-1) as triple super phosphate); NPK (NP + 75 kg K2O ha(-1) as potassium sulphate); NPKS [NPK+ S (100 kg S ha(-1))]; NPKST (NPKS + seed inoculation with Thiobacillus bacteria); NPKM (NPK + Seed inoculation with mycorrhizae fungi) and NPKSTM (NPKS + seed inoculation with Thiobacillus and mycorrhizae) and two cultivars (JK and 032). Before planting, soybean seeds were inoculated by Bradyrhizobium japonicum in all treatments. Results showed that combined inoculation of biofertilizers increased yield, however the highest yield was observed in treatment NPKST. Increasing oil content (percentage) was more pronounced in treatments NPKM, while most protein content (percentage) increasing was observed in NPKS and NPKM. Fe and Zn concentrations were unaffected significantly by fertilizer treatments, but NPKSTM showed significantly higher value of seed's Mn concentration compared to treatments NP and NPK. Although no significant difference was observed in terms ofP concentration of 032 line among fertilizer treatments, JK cultivar and NPKSTM caused a significant increasing in P concentration compared to NP, NPKS and NPKM. Present results suggested that applying biofertilizers i.e., mycorrhizae and Thiobacillus increased soybean yield compared to control (NP). Overall, this study demonstrated that soybean seed yield and its chemical composition could be affected by biofertilizer inoculation.


[Russian electrophysiotherapeutic apparatus. The problems of its design and production].
The topical organizational problems of design, production, and use of physiotherapeutical equipment (PTE) are discussed. PTE is considered to be basic tools employed in physiotherapy. The therapeutical effect of PTE on man is produced by currents of different frequency and shape by means of electrodes (which come in contact with a patient's body) or an electromagnetic field having different frequency-modulation characteristics via non-contact method by means of radiating elements. The devices which transform electromagnetic to mechanical energy may be subsumed under a separate group that includes ultrasonic therapeutical apparatuses, mechanical massage and mechanical stimulation equipment. The leading PTE designer is "VNNIMP-VITA Co. which has developed and launched about 20 varying = purpose PTE models into practice. The interest displayed by the conversed enterprises of the military industrial complex in the manufacture of medical equipment items and primarily PTE has recently caused unsystematic development of new PTE which have not been positively assessed and introduced into medical practice. New PTE models should be designed by strengthening the relations between those who design this type of medical equipment and the new structures of the Russian Academy of Medical Sciences.


Interactions of cortisol, testosterone, and resistance training: influence of circadian rhythms.
Diurnal variation of sports performance usually peaks in the late afternoon, coinciding with increased body temperature. This circadian pattern of performance may be explained by the effect of increased core temperature on peripheral mechanisms, as neural drive does not appear to exhibit nycthemeral variation. This typical diurnal regularity has been reported in a variety of physical activities spanning the energy systems, from Adenosine triphosphate-phosphocreatine (ATP-PC) to anaerobic and aerobic metabolism, and is evident across all muscle contractions (eccentric, isometric, concentric) in a large number of muscle groups. Increased nerve conduction velocity, joint suppleness, increased muscular blood flow, improvements of glycogenolysis and glycolysis, increased environmental temperature, and preferential meteorological conditions may all contribute to diurnal variation in physical performance. However, the diurnal variation in strength performance can be blunted by a repeated-morning resistance training protocol. Optimal adaptations to resistance training (muscle hypertrophy and strength increases) also seem to occur in the late afternoon, which is interesting, since cortisol and, particularly, testosterone (T) concentrations are higher in the morning. T has repeatedly been linked with resistance training adaptation, and higher concentrations appear preferential. This has been determined by suppression of endogenous production and exogenous supplementation. However, the cortisol (C)/T ratio may indicate the catabolic/anabolic environment of an organism due to their roles in protein degradation and protein synthesis, respectively. The morning elevated T level (seen as beneficial to achieve muscle hypertrophy) may be counteracted by the morning elevated C level and, therefore, protein degradation. Although T levels are higher in the morning, an increased resistance exercise-induced T response has been found in the late afternoon, suggesting greater responsiveness of the hypothalamo-pituitary-testicular axis then. Individual responsiveness has also been observed, with some participants experiencing greater hypertrophy and strength increases in response to strength protocols, whereas others respond preferentially to power, hypertrophy, or strength endurance protocols dependent on which protocol elicited the greatest T response. It appears that physical performance is dependent on a number of endogenous time-dependent factors, which may be masked or confounded by exogenous circadian factors. Strength performance without time-of-day-specific training seems to elicit the typical diurnal pattern, as does resistance training adaptations. The implications for this are (a) athletes are advised to coincide training times with performance times, and (b) individuals may experience greater hypertrophy and strength gains when resistance training protocols are designed dependent on individual T response.


Freshwater carbon and nutrient cycles revealed through reconstructed population genomes.
Although microbes mediate much of the biogeochemical cycling in freshwater, the categories of carbon and nutrients currently used in models of freshwater biogeochemical cycling are too broad to be relevant on a microbial scale. One way to improve these models is to incorporate microbial data. Here, we analyze both genes and genomes from three metagenomic time series and propose specific roles for microbial taxa in freshwater biogeochemical cycles. Our metagenomic time series span multiple years and originate from a eutrophic lake (Lake Mendota) and a humic lake (Trout Bog Lake) with contrasting water chemistry. Our analysis highlights the role of polyamines in the nitrogen cycle, the diversity of diazotrophs between lake types, the balance of assimilatory vs. dissimilatory sulfate reduction in freshwater, the various associations between types of phototrophy and carbon fixation, and the density and diversity of glycoside hydrolases in freshwater microbes. We also investigated aspects of central metabolism such as hydrogen metabolism, oxidative phosphorylation, methylotrophy, and sugar degradation. Finally, by analyzing the dynamics over time in nitrogen fixation genes and Cyanobacteria genomes, we show that the potential for nitrogen fixation is linked to specific populations in Lake Mendota. This work represents an important step towards incorporating microbial data into ecosystem models and provides a better understanding of how microbes may participate in freshwater biogeochemical cycling.


Endoscopic reconstruction of the cranial base using a pedicled nasoseptal flap.
Reconstruction of the cranial base using vascularized tissue promotes rapid and complete healing, thus avoiding complications caused by persistent communication between the cranial cavity and the sinonasal tract. The Hadad-Bassagasteguy flap (HBF), a neurovascular pedicled flap of the nasal septum mucoperiosteum and mucoperichondrium based on the nasoseptal artery, seems to be advantageous for the reconstruction of the cranial base after endonasal cranial base surgery. We performed a retrospective review of patients who underwent endonasal cranial base surgery at the University of Pittsburgh Medical Center from January 30, 2006 to January 30, 2007, identifying patients who experienced reconstruction with a vascularized septal mucosal flap (HBF). We analyzed the demographic data, pathological characteristics, site and extent of resection, use of cerebrospinal fluid (CSF) diversion techniques, and outcome. Seventy-five patients who underwent endonasal cranial base endoscopic surgery received repair with the HBF. In this population, we encountered eight postoperative CSF leaks (10.66%), all in patients who required intra-arachnoidal dissection. When we correct the statistical analysis to include only patients with intra- arachnoidal lesions, the postoperative CSF leak rate is 14.5% (eight of 55 patients). It is notable that six CSF (33%) leaks occurred in our first 25 repairs, whereas we encountered only two postoperative leaks (4%) in the last 50 patients. The corrected CSF leak rate, considering only intra-arachnoidal lesions, was two (5.4%) of 37 patients. This improvement in the CSF leak rate reflects our growing experience and comfort with this reconstructive technique. All of our failures could be matched to a specific technical mistake. In addition, we modified the flap-harvesting technique to allow for staged procedures and the removal of caudal lesions. These special circumstances require storage of the flap in the antrum during the removal of caudal lesions, and suturing of the flap in its original position for staged procedures. One patient experienced a posterior nose bleed from the posterior nasal artery. This was controlled with bipolar electrocautery, thereby preserving the flap blood supply. We encountered no infectious or wound complications in this series of patients. The donor site accumulates crusting, which requires debridement until mucosalization is complete; this usually occurs 6 to 12 weeks after surgery. The HBF is a versatile and reliable reconstructive technique for repairing defects of the anterior, middle, clival, and parasellar cranial base. Its use has resulted in a significant decrease in our incidence of CSF leaks after endonasal cranial base surgery. Attention to technical details is of paramount importance to achieve the best outcomes.


Differences in medical students' attitudes to academic misconduct and reported behaviour across the years--a questionnaire study.
This study aimed to determine attitudinal and self reported behavioural variations between medical students in different years to scenarios involving academic misconduct. A cross-sectional study where students were given an anonymous questionnaire that asked about their attitudes to 14 scenarios describing a fictitious student engaging in acts of academic misconduct and asked them to report their own potential behaviour. Dundee Medical School. Undergraduate medical students from all five years of the course. Questionnaire survey. Differences in medical students' attitudes to the 14 scenarios and their reported potential behaviour with regards to the scenarios in each of the years. For most of the scenarios there was no significant difference in the response between the years. Significant differences in the responses were found for some of the scenarios across the years, where a larger proportion of year one students regarded the scenario as wrong and would not engage in the behaviour, compared to other years. These scenarios included forging signatures, resubmitting work already completed for another part of the course, and falsifying patient information. Observed differences between the years for some scenarios may reflect a change in students' attitudes and behaviour as they progress though the course. The results may be influenced by the educational experience of the students, both in terms of the learning environment and assessment methods used. These differences may draw attention to the potential but unintentional pressures placed on medical students to engage in academic misconduct. The importance of developing strategies to engender appropriate attitudes and behaviours at the undergraduate level must be recognised.


A surge in anti-dsDNA titer predicts a severe lupus flare within six months.
Rising anti-double-stranded (ds) DNA titers have been shown by some, but not all, studies to be predictive of disease flares in systemic lupus erythematosus (SLE). We hypothesized that a rapid and substantial rise in anti-dsDNA titer (anti-dsDNA surge) would be a good predictor of a clinically important SLE flare. A matched case-control study was conducted in an academic rheumatology practice setting. Our primary endpoint was the occurrence of a severe SELENA-SLEDAI (SS) flare within six months of an anti-dsDNA surge, and secondary endpoints were mild/moderate SS flares, as well as BILAG A and B renal flares. Cases were identified as those patients whose disease course included a surge of anti-dsDNA, defined as an increase of anti-dsDNA titer by the Crithidia luciliae immunofluorescence (CLIF) assay from 0 to 3+/4+, or from 1+ to 4+, within a period of less than 12 months. The date of the anti-dsDNA surge was defined as Day 0. Two control SLE patients were identified for each case and were matched for age, sex, race, and visit date closest to case Day 0, but without an anti-dsDNA surge. Logistic regression models were used to detect associations between anti-dsDNA surges and severe SS flares. A higher proportion of cases, compared to controls, experienced a severe SS flare within six months of Day 0 (OR 6.3 (95% confidence intervals 2.0-19.9), p = 0.02). Associations with all flares and hospitalizations for flares were also observed. However, an anti-dsDNA surge was not predictive of a renal flare. An anti-dsDNA surge predicts the subsequent development of a severe SS flare within six months. Physicians should closely monitor such patients and treat promptly at the first sign of clinical activity.