Whole-brain annotation and multi-connectome cell typing quantifies circuit stereotypy in Drosophila.
The fruit fly Drosophila melanogaster combines surprisingly sophisticated behaviour with a highly tractable nervous system. A large part of the fly's success as a model organism in modern neuroscience stems from the concentration of collaboratively generated molecular genetic and digital resources. As presented in our FlyWire companion paper 1 , this now includes the first full brain connectome of an adult animal. Here we report the systematic and hierarchical annotation of this ~130,000-neuron connectome including neuronal classes, cell types and developmental units (hemilineages). This enables any researcher to navigate this huge dataset and find systems and neurons of interest, linked to the literature through the Virtual Fly Brain database 2 . Crucially, this resource includes 4,552 cell types. 3,094 are rigorous consensus validations of cell types previously proposed in the hemibrain connectome 3 . In addition, we propose 1,458 new cell types, arising mostly from the fact that the FlyWire connectome spans the whole brain, whereas the hemibrain derives from a subvolume. Comparison of FlyWire and the hemibrain showed that cell type counts and strong connections were largely stable, but connection weights were surprisingly variable within and across animals. Further analysis defined simple heuristics for connectome interpretation: connections stronger than 10 unitary synapses or providing >1% of the input to a target cell are highly conserved. Some cell types showed increased variability across connectomes: the most common cell type in the mushroom body, required for learning and memory, is almost twice as numerous in FlyWire as the hemibrain. We find evidence for functional homeostasis through adjustments of the absolute amount of excitatory input while maintaining the excitation-inhibition ratio. Finally, and surprisingly, about one third of the cell types proposed in the hemibrain connectome could not yet be reliably identified in the FlyWire connectome. We therefore suggest that cell types should be defined to be robust to inter-individual variation, namely as groups of cells that are quantitatively more similar to cells in a different brain than to any other cell in the same brain. Joint analysis of the FlyWire and hemibrain connectomes demonstrates the viability and utility of this new definition. Our work defines a consensus cell type atlas for the fly brain and provides both an intellectual framework and open source toolchain for brain-scale comparative connectomics.


Sympathetically and nonsympathetically mediated onset of excess fat deposition in Zucker rats.
To evaluate whether a low sympathetic activity is essential for the onset of Zucker rat obesity, we artificially reared fa/fa and Fa/fa littermates from 4 days of age with identical amounts of milk containing norepinephrine (NE). The NE concentration was adjusted to keep the mass-specific metabolic rate (MR) roughly either 100 or 50% above its thermoneutral level. At both doses, no genotype difference in total MR occurred before day 16, but between 16 and 21 days of age, total MR of fa/fa pups was 10% less than that of their Fa/fa littermates. The 21-day-old fa/fa pups had approximately 1 g more body fat than their lean littermates, and this was energetically accounted for to 50-80% by their lower MR and to 10-25% by the reduced growth of their fat-free dry mass. We conclude that factors not dependent on decreased sympathetic outflow trigger the onset of excess fat deposition in the 3rd postnatal wk, even if sympathetic activity was high throughout the previous weeks during which excessive fat deposition is mainly dependent on decreased thermoregulatory thermogenesis.


Reliability and validity of the Short Portable Mental Status Questionnaire administered by telephone.
Effective, economical, and reliable means of screening subjects for cognitive impairment when personal contact is not feasible could facilitate epidemiologic studies and longitudinal assessment. The Short Portable Mental Status Questionnaire (SPMSQ) is a 10-item examination that has been found reliable and valid in distinguishing demented subjects from cognitively intact subjects when given face to face. The current study assessed the utility of a telephone version of the SPMSQ in patients evaluated in an outpatient geriatric assessment program. Mean scores for both test versions decreased with dementia severity and correlated significantly. Mean score differences between the two versions were not affected by reports of hearing impairment or the time interval between test administration. Both test versions correlated significantly with the Mini-Mental State Examination. In distinguishing demented from nondemented subjects, sensitivity and specificity were .74 and .79 for the telephone test and .74 and .91 for the face-to-face test, respectively.


Modelling the decline and future of hepatitis A transmission in Australia.
Hepatitis A incidence has declined in most countries through a combination of prevention measures, augmented through the use of a highly effective vaccine. In Australia, the proportion of the population susceptible to hepatitis A infection has declined over time due to high rates of opportunistic vaccination as well as the sustained inflow of seropositive immigrants from high-endemicity countries. These factors have contributed to a rapid decline in incidence. An age-structured hepatitis A transmission model incorporating demographic changes was fitted to seroprevalence and disease notification data and used to project incidence trends and transmission potential for hepatitis A in the general population. Robustness of findings was assessed through worst-case scenarios regarding vaccine uptake, migration and the duration of immunity. The decline in age-specific seroprevalence until the introduction of hepatitis A vaccine in 1994 was well explained through a declining basic reproduction number (R0 ) that remained >1. Accounting for existing immunity, we estimated that the effective reproduction number (Reff ) <1 in the general population of Australia since the early 1990s, declining more rapidly after the introduction of the hepatitis A vaccine. Future projections under a variety of scenarios support Reff remaining <1 with continued low incidence in the general population. In conclusion, our results suggest that sustained endemic transmission in the general Australian population is no longer possible although risks of sporadic outbreaks remain. This suggests potential for local elimination of hepatitis A infection in Australia, provided that elimination criteria can be defined and satisfied in risk groups. The methodology used here to investigate elimination potential can easily be replicated in settings such as in the USA where sequential seroprevalence studies are supported by routine notification data.


Identifying a set that contains the best dynamic treatment regimes.
A dynamic treatment regime (DTR) is a treatment design that seeks to accommodate patient heterogeneity in response to treatment. DTRs can be operationalized by a sequence of decision rules that map patient information to treatment options at specific decision points. The sequential, multiple assignment, randomized trial (SMART) is a trial design that was developed specifically for the purpose of obtaining data that informs the construction of good (i.e. efficacious) decision rules. One of the scientific questions motivating a SMART concerns the comparison of multiple DTRs that are embedded in the design. Typical approaches for identifying the best DTRs involve all possible comparisons between DTRs that are embedded in a SMART, at the cost of greatly reduced power to the extent that the number of embedded DTRs (EDTRs) increase. Here, we propose a method that will enable investigators to use SMART study data more efficiently to identify the set that contains the most efficacious EDTRs. Our method ensures that the true best EDTRs are included in this set with at least a given probability. Simulation results are presented to evaluate the proposed method, and the Extending Treatment Effectiveness of Naltrexone SMART study data are analyzed to illustrate its application.


Proposing a method of regional assessment and a novel outcome measure in rheumatoid arthritis.
We proposed a method of regional assessment in patients with rheumatoid arthritis. The utility of this method was demonstrated by assessing drug efficacy in patients who received infliximab (n = 31) or tocilizumab (n = 6). Joints were divided into four regions: upper/large, upper/small, lower/large, and lower/small. The total joint index was calculated as follows: the sum of tender and swollen joint counts divided by the number of evaluable joints in each region. At the baseline, the total joint index of the upper/small region was the lowest and that of the lower/large region was the highest compared with other regions. The change in the total joint index from the baseline to the 30-week point (Δ) did not differ among the four regions. There were significant close relations of Δ between the upper/small and the upper/large region and between the lower/small and the lower/large region. This method allows us to focus on a specific region and to compare and contrast among them.


Temporal Dynamic of the Ratio between Monopartite Begomoviruses and Their Associated Betasatellites in Plants, and Its Modulation by the Viral Gene βC1.
The begomovirus-betasatellite complex constantly threatens crops in Asia. However, the quantitative relationship between begomoviruses and betasatellites remains largely unknown. The quantities of tobacco curly shoot virus (TbCSV) and its betasatellite (TbCSB) and their ratio varied significantly in initial infection, and thereafter, the ratio tended to become constant. The TbCSB/TbCSV ratio in agrobacteria inoculum significantly affected that in plants in the initial infection but not thereafter. Null-mutation of βC1 that encodes a multifunctional protein important for pathogenesis in TbCSB significantly reduced the TbCSB/TbCSV ratio in plants. Viral inoculum plants with higher TbCSB/TbCSV ratios promoted whitefly transmission of the virus. The expression of AV1 encoded by TbCSV, βC1 encoded by TbCSB and the βC1/AV1 ratio varied significantly in the initial infection and thereafter the ratio tended to become constant. Additionally, the temporal dynamics of the ratio between another begomovirus and its betasatellite was similar to that of TbCSV and was positively regulated by βC1. These results indicate that the ratio between monopartite begomoviruses and betasatellites tend to become constant as infection progresses, and is modulated by βC1, but a higher betasatellite/begomovirus ratio in virally inoculated plants promotes virus transmission by whiteflies. Our findings provide novel insights into the association between begomoviruses and betasatellites.


[Selective estrogen receptors modulators (SERMs): biochemistry, pharmacology, and clinical use in gynecology].
The selective strogen receptors modulators (SERMs) were initially developed as antistrogens for the treatment of breast cancer, but their unusual properties have led to their use in the treatment and prevention of other diseases as well. SERMs bind the strogen receptor (ER) and modulated ER-mediated gene transcription. Tamoxifen is an effective treatment for hormone responsive breast cancer and can prevent breast cancer in high-risk women. Raloxifene was approved for the prevention and treatment of osteoporosis in postmenopausal women, also appears to prevent breast cancer. Other SERMs are in development, with the goal of improving efficacy and reducing toxicity.


Iranian scientists and French showers: collaborative fact-checking of identity-salient online information.
In this study, we investigate what leads people to fact-check online information, how they fact-check such information in practice, how fact-checking affects their judgments about the information's credibility, and how each of the above processes is affected by the salience of the information to readers' cultural identities. Eight pairs of adult participants were recruited from diverse cultural backgrounds to participate online in joint fact-checking of suspect Tweets. To examine their collaborative deliberations we developed a novel experimental design and analytical model. Our analyses indicate that the salience of online information to people's cultural identities influences their decision to fact-check it, that fact-checking deliberations are often non-linear and iterative, that collaborative fact-checking leads people to revise their initial judgments about the credibility of online information, and that when online information is highly salient to people's cultural identities, they apply different standards of credibility when fact-checking it. In conclusion, we propose that cultural identity is an important factor in the fact-checking of online information, and that joint fact-checking of online information by people from diverse cultural backgrounds may have significant potential as an educational tool to reduce people's susceptibility to misinformation.


Barriers to the Integration of Care in Inter-Organisational Settings: A Literature Review.
In recent years, inter-organisational collaboration between healthcare organisations has become of increasingly vital importance in order to improve the integration of health service delivery. However, different barriers reported in academic literature seem to hinder the formation and development of such collaboration. This systematic literature review of forty studies summarises and categorises the barriers to integrated care in inter-organisational settings as reported in previous studies. It analyses how these barriers operate. Within these studies, twenty types of barriers have been identified and then categorised in six groups (barriers related to administration and regulation, barriers related to funding, barriers related to the inter-organisational domain, barriers related to the organisational domain, barriers related to service delivery, and barriers related to clinical practices). Not all of these barriers emerge passively, some are set up intentionally. They are not only context-specific, but are also often related and influence each other. The compilation of these results allows for a better understanding of the characteristics and reasons for the occurrence of barriers that impede collaboration aiming for the integration of care, not only for researchers but also for practitioners. It can help to explain and counteract the slow progress and limited efficiency and effectiveness of some of the inter-organisational collaboration in healthcare settings.


Chemical (VX) terrorist threat: public knowledge, attitudes, and responses.
This article reports the results of a study of people's perceptions and reactions to a hypothetical terrorist attack involving a chemical agent (specifically, the nerve agent VX). Thirteen focus groups composed of 8 to 12 participants each were conducted using trained moderators. To achieve a broad representation of perspectives, the groups were conducted in several regions and included urban and rural locations. In addition, a variety of population groups, such as African Americans, Hispanics, American Indians, Asians, and people with English as a second language, were included in the study. Findings demonstrated fear, fatalism, and unfulfilled information needs related to the threat agent. To better prepare the public for VX threats or threats from other highly toxic chemical agents, it will be important to emphasize that VX exposure can be avoided or reduced, that VX effects can be treated, and that VX can be survived if appropriate protective measures are taken. Related findings from the focus groups are that participants preferred television, radio, and the Emergency Alert System for emergency messages and that people prefer to hear information about a chemical attack from a well-known, well-respected public figure or from a content expert on chemical attacks, protective actions, and health. In addition, local television meteorologists were identified as a category of trusted conveyers of important information in relation to chemical terrorist attacks.


Light-Controlled Lipid Interaction and Membrane Organization in Photolipid Bilayer Vesicles.
Controlling lateral interactions between lipid molecules in a bilayer membrane to guide membrane organization and domain formation is a key factor for studying and emulating membrane functionality in synthetic biological systems. Here, we demonstrate an approach to reversibly control lipid organization, domain formation, and membrane stiffness of phospholipid bilayer membranes using the photoswitchable phospholipid azo-PC. azo-PC contains an azobenzene group in the sn2 acyl chain that undergoes reversible photoisomerization on illumination with UV-A and visible light. We demonstrate that the concentration of the photolipid molecules and also the assembly and disassembly of photolipids into lipid domains can be monitored by UV-vis spectroscopy because of a blue shift induced by photolipid aggregation.


High-permeability pulmonary edema: nursing assessment, diagnosis, and interventions.
High-permeability pulmonary edema (HPPE) is a problem affecting 150,000 to 200,000 critically ill patients yearly. In HPPE the alveolar-capillary membrane is injured. The resulting increased permeability of the alveolar-capillary membrane allows shifts of fluid and protein into the interstitial fluid space and alveolus. As hypoxemia develops, the nurse assesses cardinal signs and symptoms derived from the physical examination and observations. Clinical data consisting of results from various laboratory and diagnostic studies confirm the diagnosis of HPPE. Finally, nursing diagnoses can be delineated as the basis on which expert nursing care is planned and implemented.


Anterior hypopituitarism in adult survivors of childhood cancers treated with cranial radiotherapy: a report from the St Jude Lifetime Cohort study.
To estimate the prevalence of and risk factors for growth hormone deficiency (GHD), luteinizing hormone/follicle-stimulating hormone deficiencies (LH/FSHD), thyroid-stimulatin hormone deficiency (TSHD), and adrenocorticotropic hormone deficiency (ACTHD) after cranial radiotherapy (CRT) in childhood cancer survivors (CCS) and assess the impact of untreated deficiencies. Retrospective study in an established cohort of CCS with 748 participants treated with CRT (394 men; mean age, 34.2 years [range, 19.4 to 59.6 years] observed for a mean of 27.3 years [range, 10.8 to 47.7 years]). Multivariable logistic regression was used to study associations between demographic and treatment-related risk factors and pituitary deficiencies, as well as associations between untreated deficiencies and cardiovascular health, bone mineral density (BMD), and physical fitness. The estimated point prevalence was 46.5% for GHD, 10.8% for LH/FSHD, 7.5% for TSHD, and 4% for ACTHD, and the cumulative incidence increased with follow-up. GHD and LH/FSHD were not treated in 99.7% and 78.5% of affected individuals, respectively. Male sex and obesity were significantly associated with LH/FSHD; white race was significant associated with LH/FSHD and TSHD. Compared with CRT doses less than 22 Gy, doses of 22 to 29.9 Gy were significantly associated with GHD; doses ≥ 22 Gy were associated with LH/FSHD; and doses ≥ 30 Gy were associated with TSHD and ACTHD. Untreated GHD was significantly associated with decreased muscle mass and exercise tolerance; untreated LH/FSHD was associated with hypertension, dyslipidemia, low BMD, and slow walking; and both deficits, independently, were associated with with abdominal obesity, low energy expenditure, and muscle weakness. Anterior pituitary deficits are common after CRT. Continued development over time is noted for GHD and LH/FSHD with possible associations between nontreatment of these conditions and poor health outcomes.


The incidence of PAX6 mutation in patients with simple aniridia: an evaluation of mutation detection in 12 cases.
Twelve aniridia patients, five with a family history and seven presumed to be sporadic, were exhaustively screened in order to test what proportion of people with aniridia, uncomplicated by associated anomalies, carry mutations in the human PAX6 gene. Mutations were detected in 90% of the cases. Three mutation detection techniques were used to determine if one method was superior for this gene. The protein truncation test (PTT) was used on RT-PCR products, SSCP on genomic PCR amplifications, and chemical cleavage of mismatch on both RT-PCR and genomic amplifications. For RT-PCR products, only the translated portion of the gene was screened. On genomic products exons 1 to 13 (including 740 bp of the 3' untranslated sequence and all intron/exon boundaries) were screened, as was a neuroretina specific enhancer in intron 4. Ten of the possible 12 mutations in the five familial cases and five of the sporadic patients were found, all of which conformed to a functional outcome of haploinsufficiency. Five were splice site mutations (one in the donor site of intron 4, two in the donor site of intron 6, one in each of the acceptor sites of introns 8 and 9) and five were nonsense mutations in exons 8, 9, 10, 11, and 12. SSCP analysis of individually amplified exons, with which nine of the 10 mutations were seen, was the most useful detection method for PAX6.


Recovery of semivolatile organic compounds during sample preparation: implications for characterization of airborne particulate matter.
Semivolatile compounds present special analytical challenges not met by conventional methods for analysis of ambient particulate matter (PM). Accurate quantification of PM-associated organic compounds requires validation of the laboratory procedures for recovery over a wide volatility and polarity range. To meet these challenges, solutions of n-alkanes (nC12-nC40) and polycyclic aromatic hydrocarbons PAHs (naphthalene to benzo[ghi]perylene) were reduced in volume from a solvent mixture (equal volumes of hexane, dichloromethane and methanol), to examine recovery after reduction in volume. When the extract solution volume reached 0.5 mL the solvent was entirely methanol, and the recovery averaged 60% for n-alkanes nC12-nC25 and PAHs from naphthalene to chrysene. Recovery of higher MW compounds decreased with MW, because of their insolubility in methanol. When the walls of the flasks were washed with 1 mL of equal parts hexane and dichloromethane (to reconstruct the original solvent composition), the recovery of nC18 and higher MW compounds increased dramatically, up to 100% for nC22-nC32 and then slowly decreasing with MW due to insolubility. To examine recovery during extraction of the components of the High Capacity Integrated Gas and Particle Sampler, the same standards were used to spike its denuders and filters. For XAD-4 coated denuders and filters, normalized recovery was >95% after two extractions. Recovery from spiked quartz filters matched the recovery from the coated surfaces for alkanes nC18 and larger and for fluoranthene and larger PAHs. Lower MW compounds evaporated from the quartz filter with the spiking solvent. This careful approach allowed quantification of organics by correcting for volatility- and solubility-related sample preparation losses. This method is illustrated for an ambient sample collected with this sampler during the Texas Air Quality Study 2000.


Effect of the addition of live yeast (Saccharomyces cerevisiae) on growth and carcass quality of steers fed high-forage or high-grain diets and on feed digestibility and in situ degradability.
The effects of adding yeast culture (Saccharomyces cerevisiae, 5 x 10(9) live organisms/g of growth medium) at 10 g/steer daily to three diets consisting of 75% alfalfa silage and 25% barley, 96% corn silage and 4.0% soybean meal, or 75% dry-rolled barley and 25% alfalfa hay on performance of growing and finishing steers and carcass characteristics, feed digestibility, and degradability in the rumen were determined. In separate trials over 2 yr, the performance of steers fed the three diets with no supplementary live yeast (control; C) or with live yeast (Y) was determined using 72 Hereford steer calves in eight pens each year in a randomized complete block design. The diets were fed sequentially and the steers were randomized before diets were switched. Each year at the end of the high-grain feeding segment of the trial, the steers were slaughtered when ultrasound backfat was such that the carcass would grade A1 or A2 (Canadian grades). The digestibility of feed DM, CP, and NDF of the three diets was determined sequentially using six and eight mature, ruminally cannulated steers in 2 yr, respectively. The degradation characteristics of the three diets in the rumen were determined using the nylon bag technique in two steers for each additive treatment. In the 1st yr, the feed efficiency of the corn silage diet was lower (P < .05) for C than for Y steers (5.9 vs 6.8). Differences (P < .05) were not observed between C and Y steers for any of the performance or carcass characteristics.(ABSTRACT TRUNCATED AT 250 WORDS)


Validation of reference housekeeping genes for gene expression studies in western corn rootworm (Diabrotica virgifera virgifera).
Quantitative Real-time PCR (qRT-PCR) is a powerful technique to investigate comparative gene expression. In general, normalization of results using a highly stable housekeeping gene (HKG) as an internal control is recommended and necessary. However, there are several reports suggesting that regulation of some HKGs is affected by different conditions. The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is a serious pest of corn in the United States and Europe. The expression profile of target genes related to insecticide exposure, resistance, and RNA interference has become an important experimental technique for study of western corn rootworms; however, lack of information on reliable HKGs under different conditions makes the interpretation of qRT-PCR results difficult. In this study, four distinct algorithms (Genorm, NormFinder, BestKeeper and delta-CT) and five candidate HKGs to genes of reference (β-actin; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; β-tubulin; RPS9, ribosomal protein S9; EF1a, elongation factor-1α) were evaluated to determine the most reliable HKG under different experimental conditions including exposure to dsRNA and Bt toxins and among different tissues and developmental stages. Although all the HKGs tested exhibited relatively stable expression among the different treatments, some differences were noted. Among the five candidate reference genes evaluated, β-actin exhibited highly stable expression among different life stages. RPS9 exhibited the most similar pattern of expression among dsRNA treatments, and both experiments indicated that EF1a was the second most stable gene. EF1a was also the most stable for Bt exposure and among different tissues. These results will enable researchers to use more accurate and reliable normalization of qRT-PCR data in WCR experiments.


The Complete Blood Count: A Practical Tool for the Pediatrician.
Every child should have a baseline hematology evaluation with hemoglobin and hematocrit levels starting at age 12 months, or younger if clinically indicated. Although history and physical examination provide key information needed to diagnose blood disorders, the addition of a complete blood count (CBC) with differential count and reticulocyte count allows the clinician to narrow the differential diagnosis and tailor the subsequent evaluation. The interpretation of CBC results is a skill that requires practice. Every clinician can learn to identify possible diagnoses before consulting a specialist. This review provides a step-by-step approach for CBC interpretation with tools to help the clinician diagnose and interpret the most common blood disorders seen in the general pediatric clinic or inpatient setting.


Myxoma virus virotherapy for glioma in immunocompetent animal models: optimizing administration routes and synergy with rapamycin.
Oncolytic myxoma virus (MYXV) is being developed as a novel virotherapeutic against human brain cancer and has promising activity against human brain tumor models in immunocompromised hosts. Because an intact immune system could reduce its efficacy, the purpose of this study was to evaluate the oncolytic potential of MYXV in immunocompetent racine glioma models. Here, we report that MYXV infects and kills all racine cell glioma lines and that its effects are enhanced by rapamycin. Intratumoral administration of MYXV with rapamycin improved viral replication in the tumor and significantly prolonged host survival. Similarly, coadministration via a method of convection-enhanced delivery (CED) enhanced viral replication and efficacy in vivo. Mechanisms by which rapamycin improved MYXV oncolysis included an inhibition of type I IFN production in vitro and a reduction of intratumoral infiltration of CD68(+) microglia/macrophages and CD163(+) macrophages in vivo. Our findings define a method to improve MYXV efficacy against gliomas by rapamycin coadministration, which acts to promote immune responses engaged by viral delivery.


Influence of fumigants on soil microbial diversity and survival of E. coli O157:H7.
The aim of this study was to assess the effects of soil fumigation with methyl bromide (MeBr; CH(3)Br) and methyl iodide (MeI, iodomethane; CH(3)I) on the microbial community structure and diversity in two soils and determine the effects of microbial diversity on the survival of Escherichia coli O157:H7 from contaminated irrigation water. Polymerase chain reaction (PCR) was used to amplify 16S rRNA from total bacterial community composition and the products were subjected to denaturing gradient gel electrophoresis (DGGE). The Shannon-Weaver index of diversity (H') was used to determine the effects of both fumigants on soil microbial diversity. The effect was more severe in sandy soil than in clay soil at the normal application rate of MeBr and MeI. Our results showed that MeBr and MeI have about the same effects on soil microbial diversity. The two fumigants had greater impact on microbial diversity in sandy soil than in clay soil and this resulted in higher survival of E. coli O157:H7 in sandy soil than in clay soil during the 50 days that the study was conducted. MeBr has been used as soil fumigant for >40 years with no serious detrimental effects on agricultural production and our research also suggests that the use of MeI may also produce no long-term detrimental effects on agricultural production since both fumigants had about the same effects on soil microbial communities. Therefore, soil systems with reduced microbial diversity may offer greater opportunities for the survival of pathogenic bacteria such as E. coli O157:H7.


Evaluation of Bilateral Asymmetry in the Humerus of Human Skeletal Specimen.
Several studies have established a relationship between morphological and behavioral asymmetry making investigations of bilateral bone asymmetry an attractive and important research area. The purpose of this study was to investigate bilateral asymmetry patterns of skeletal specimen from five geographical locations (Rwanda, Burundi, Congo, Kenya, and Uganda) at Galloway Osteological Collection, Department of Anatomy, School of Biomedical Sciences, Makerere University College of Health Sciences. The angle of torsion and retroversion, mid-shaft circumference, length, and weight of 232 pairs of humeri were determined. A Torsiometer was used to measure the angle of torsion in degrees according to Krahl and Evans 1945, a tape was used to measure the mid-shaft circumference at the level of the apex of the deltoid V, and the length in cm was determined. An osteometric board was used to measure the length of the humerus in centimeters. A weighing balance was used to measure the weight of the humerus in grams. The analysis of humeral asymmetry with respect to parameters of the human skeletal specimen at the Galloway Osteological Collection Mulago revealed bilateral asymmetrical status observed in the angle of torsion, length, weight, and mid-shaft circumference. Our result mostly showed lateralization to the right in all the parameters investigated except the torsion angle which is to the left. Our investigation revealed that humeral torsion is inversely proportional to weight, length, and mid-shaft circumference of the humerus. This study established the existence of bilateral asymmetries in the humeri of all the geographical regions investigated with more asymmetry observed in the male compared with the female.


[Adrenergic mechanism of the stimulation of the small intestine motor activity].
Myoelectrical activity of the duodenum and ileum was recorded in rabbits with implanted electrode. Acute haemorrhage caused either elevation or reduction of the firing rate, the responses being abolished with a combined blockade of alpha- and beta-adrenoreceptors. Beta-adrenoreceptors agonist isoproterenol resulted in a biphasic (decrease-increase) change of the myoelectrical activity which could be abolished with blockade of beta-adrenoreceptors. The character of the small intestine motor responses to acute haemorrhage and isoproterenol infusion depended on the initial level of the intestinal contractile activity. The findings suggest that catecholamines induce a small intestinal motor effect via stimulation of both inhibitory adrenoreceptors and excitatory presynaptic adrenoreceptors.


Conjunctival squamous intraepithelial neoplasia and its differential diagnosis.
Conjunctival squamous intraepithelial neoplasia (CSIN) represents the in situ precursor of squamous cell carcinoma. The graded severity of intraepithelial dysplasia is considered a measure of risk for progression to invasive carcinoma. The range of cytoarchitectural changes in CSIN overlaps those of reactive atypia, squamous epithelial papilloma and in situ sebaceous carcinoma. Pseudoepitheliomatous hyperplasia and benign hereditary dyskeratosis of the conjunctiva are conditions without risk of neoplastic transform that are potentially mistaken for CSIN.


Preferential expression of one beta-tubulin gene during flagellate development in Physarum.
The microbial eukaryote Physarum polycephalum displays several distinct cell types in its life cycle, including amoebae, flagellates and plasmodia. Despite its relative simplicity, Physarum has a tubulin gene family of complexity comparable to that of Drosophila. We have identified beta-tubulin cDNAs from Physarum that are derived from the betA beta-tubulin locus and encode beta 1A tubulin. We have also identified a partial cDNA for the unlinked betB beta-tubulin gene, which encodes beta 1B tubulin. The polypeptide sequences encoded by betA and betB show 99% identity, but the nucleotide sequences show only 85% identity, consistent with an ancient duplication of these genes. The betB gene is expressed in amoebae, flagellates and plasmodia, whereas betA is expressed only in amoebae and flagellates. During the amoeba-flagellate transition the level of betA transcript increases over 100-fold, while the level of betB transcript changes very little. Thus Physarum has a mechanism for regulating the level of discrete beta-tubulin transcripts differentially during flagellate development. A need for this differential regulation could account for the maintenance of the virtually isocoding betA and betB beta-tubulin genes.


Decadal shifts in traits of reef fish communities in marine reserves.
Marine reserves are known to impact the biomass, biodiversity, and functions of coral reef fish communities, but the effect of protective management on fish traits is less explored. We used a time-series modelling approach to simultaneously evaluate the abundance, biomass, and traits of eight fish families over a chronosequence spanning 44 years of protection. We constructed a multivariate functional space based on six traits known to respond to management or disturbance and affect ecosystem processes: size, diet, position in the water column, gregariousness, reef association, and length at maturity. We show that biomass increased with a log-linear trend over the time-series, but abundance only increased after 20 years of closure, and with more variation among reserves. This difference is attributed to recovery rates being dependent on body sizes. Abundance-weighted traits and the associated multivariate space of the community change is driven by increased proportions over time of the trait categories: 7-15 cm body size; planktivorous; species low in the water column; medium-large schools; and species with high levels of reef association. These findings suggest that the trait compositions emerging after the cessation of fishing are novel and dynamic.


A Quality Improvement Project to Reduce Combination Acetaminophen-opioid Prescriptions to Pediatric Orthopedic Patients.
Acetaminophen-opioid analgesics are among the most commonly prescribed pain medications in pediatric orthopedic patients. However, these combined opioid analgesics do not allow for individual medication titration, which can increase the risk of opioid misuse and hepatoxicity from acetaminophen. The primary aim of this quality improvement project was to alter the prescribing habits of pediatric orthopedic providers at our institution from postoperative acetaminophen-opioid analgesics to independent acetaminophen and opioids. The study took place in a level 1 trauma center at a children's hospital. A multidisciplinary team of health professionals utilized lean methodology to develop a project plan. Guided by a key driver diagram, we removed acetaminophen-oxycodone products from hospital formulary, implemented a revised inpatient and outpatient electronic order set, and conducted multiple education efforts. Outcomes included inpatient and outpatient percent combined acetaminophen-opioid orders by surgical providers over 27 months. Before the intervention, inpatient acetaminophen-opioid products accounted for an average of 46% of all opioid prescriptions for orthopedic patients. After the intervention and multiple educational efforts, we reported a reduction in the acetaminophen-opioid products to 2.9%. For outpatient prescriptions, combined analgesics accounted for 88% before the intervention, and we reported a reduction to 15% after the intervention. By removing acetaminophen-oxycodone products from hospital formulary, educating the medical staff, and employing revised electronic order sets, the prescribing practice of pediatric orthopedic surgeons changed from the routine use of acetaminophen-opioid analgesics to independent medications.


Intracutaneous distributions of fluconazole, itraconazole, and griseofulvin in Guinea pigs and binding to human stratum corneum.
We have compared the distribution of fluconazole (FLC) with that of itraconazole (ITC) and griseofulvin (GRF) in the abdominal skin tissues after a single oral dose was administered to guinea pigs. The FLC concentrations in the stratum corneum reached a peak at 2 h after administration and were similar to those of ITC and higher than those of GRF in spite of the administration of a lower dose. GRF was eliminated from the stratum corneum faster than FLC and ITC. The FLC concentrations were also remarkably higher than those of ITC and GRF in the epidermis-cutis but lower in the subcutaneous fatty tissue. The distribution characteristics of each drug result from differences in their physicochemical properties. Following the administration of multiple doses, the FLC concentrations in the stratum corneum were highest in the abdominal skin tissues; those at 24 h after each administration increased gradually and were maintained at a level more than 10 times higher than that of the plasma concentrations. The FLC concentrations in the planta pedis stratum corneum and in the nail showed good dose proportionality and obvious accumulation and were 60 and 40 times as high as that in plasma on day 14. The extent of binding of FLC to human corneous keratin in vitro was about 10%, which is lower than those of ITC (94 to 97%) and GRF (36 to 38%). FLC, unlike ITC, therefore, is presumed to exist in the stratum corneum at high concentrations in an active nonbinding form. These excellent intracutaneous pharmacokinetic properties of FLC probably account in large part for the in vivo efficacy of FLC.


Macrorhabdus ornithogaster (Megabacterium) infection in adult hobby chickens in North America.
A field investigation was conducted on a flock of adult hobby chickens showing intermittent signs of enteritis. Roosters examined in the initial field visit and postmortem had cecal worms, roundworms, tetratrichomonads, and coccidiosis. Macrorhabdus ornithogaster was diagnosed histologically in the mucosal isthmus of the proventriculus and ventriculus. Three roosters and two hens were examined in a follow-up investigation of the flock conducted 9 days later. Macrorhabdus ornithogaster was confirmed in one hen.


Activation and phosphorylation of the 'dense-vesicle' high-affinity cyclic AMP phosphodiesterase by cyclic AMP-dependent protein kinase.
Incubation of a hepatocyte particulate fraction with ATP and the isolated catalytic unit of cyclic AMP-dependent protein kinase (A-kinase) selectively activated the high-affinity 'dense-vesicle' cycle AMP phosphodiesterase. Such activation only occurred if the membranes had been pre-treated with Mg2+. Mg2+ pre-treatment appeared to function by stimulating endogenous phosphatases and did not affect phosphodiesterase activity. Using the antiserum DV4, which specifically immunoprecipitated the 51 and 57 kDa components of the 'dense-vesicle' phosphodiesterase from a detergent-solubilized membrane extract, we isolated a 32P-labelled phosphoprotein from 32P-labelled hepatocytes. MgCl2 treatment of such labelled membranes removed 32P from the immunoprecipitated protein. Incubation of the Mg2+-pre-treated membranes with [32P]ATP and A-kinase led to the time-dependent incorporation of label into the 'dense-vesicle' phosphodiesterase, as detected by specific immunoprecipitation with the antiserum DV4. The time-dependences of phosphodiesterase activation and incorporation of label were similar. It is suggested (i) that phosphorylation of the 'dense-vesicle' phosphodiesterase by A-kinase leads to its activation, and that such a process accounts for the ability of glucagon and other hormones, which increase intracellular cyclic AMP concentrations, to activate this enzyme, and (ii) that an as yet unidentified kinase can phosphorylate this enzyme without causing any significant change in enzyme activity but which prevents activation and phosphorylation of the phosphodiesterase by A-kinase.


Postoperative condition after the use of remifentanil with a small dose of piritramide compared with a fentanyl-based protocol in patients undergoing craniotomy.
The use of remifentanil requires other analgesics for postoperative pain relief compared to fentanyl in patients undergoing craniotomy. This could possibly reduce the postoperative advantages of this short-acting opioid. We compared remifentanil and fentanyl-based anaesthesia in a randomized observer and patient blinded trial on patients, undergoing an elective craniotomy. Twenty patients received anaesthesia using remifentanil with a small dose of piritramide (0.1 mg kg(-1)) after closure of the dura mater. Twenty patients underwent a fentanyl-based protocol. In both groups, anaesthesia was induced with thiopental and rocuronium, and maintained with 0.6-1 minimum alveolar concentration (MAC) isoflurane in a nitrous oxide/oxygen mixture 2:1 and rocuronium. Patients received 1 g of paracetamol rectally postoperatively. A visual analogue scale (VAS) for pain, the Glasgow Coma Score, a modified Aldrete Score, arterial carbon dioxide tension (PaCO2) and piritramide consumption were evaluated every half an hour postoperatively. No significant differences were found for pain, Aldrete or Glasgow Coma scores or for PaCO2 between the groups when controlled for age, although the pain and Glasgow Coma Scores were consistently higher and PaCO2 lower in the remifentanil group. Furthermore, 11 out of 20 patients in the remifentanil group requested extra piritramide as opposed to 7 out of 20 in the fentanyl group (P = 0.11). Despite the intraoperative use of piritramide in the remifentanil group, patients experienced more pain postoperatively. A significant influence of age on pain intensity was found. The use of remifentanil with a small dose of piritramide of 0.1 mg kg(-1) has no evident advantage over the use of fentanyl considering the postoperative conditions after craniotomy.


Effects of Gene Polymorphisms, Metabolic Activity, and Content of Alcohol Dehydrogenase and Acetaldehyde Dehydrogenases on Prognosis of Hepatocellular Carcinoma Patients.
Alcohol dehydrogenase and acetaldehyde dehydrogenases have been associated with hepatocellular carcinoma, but how alcohol dehydrogenase and acetaldehyde dehydrogenases alter the prognosis of hepatocellular carcinoma have not been completely elucidated. Metabolic activities, gene polymorphisms, and content of alcohol dehydrogenase and acetaldehyde dehydrogenases were determined in 68 fibrotic livers from hepatocellular carcinoma patients. These characteristics were then correlated with clinical features and prognosis in these patients. The median survival time of the ALDH-high activity group (727 days) was increased by 128% compared with that of ALDH-low activity group (319 days), and there was a significant negative correlation between the activity of acetaldehyde dehydrogenases and the level of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase. There was no difference in survival time between ALDH2-high and ALDH2-low expression group, though the activity of acetaldehyde dehydrogenases had correlation with the content of ALDH2 (r = 0.6887, P < .001). Mutation at ALDH2rs671 significantly decreased both the activity and content of acetalde- hyde dehydrogenases, but the polymorphism had no relationship with progression of hepatocellular carcinoma patients. In addition, the activity and 3 polymorphisms of alcohol dehydrogenase had no effect on overall survival. Mutation at ADH1Crs698 significantly decreased both the activity and content of alcohol dehydrogenase (P < .05), mutation at ADH1C rs2241894 had an inverse effect, and mutation at ADH1B rs1229984 increased activity but did not affect content. The activity of alcohol dehydrogenase had a moderate cor- relation with the content of ADH1A and ADH1C in livers (P < .05). Low activity of acetaldehyde dehydrogenases in livers correlates with poor prognosis and clinical progression in hepatocel- lular carcinoma patients, and both gene polymorphisms and content influence its metabolic activity.


Methotrexate-based ionic liquid as a potent anticancer drug for oral delivery: In vivo pharmacokinetics, biodistribution, and antitumor efficacy.
Oral delivery of the sparingly soluble drug methotrexate (MTX) is challenging owing to its poor bioavailability and low solubility. To address this challenge, the present study reports the conversion of MTX into a series of five ionic liquids (ILs) comprising a cationic component-i.e., cholinium (Cho), tetramethylammonium (TMA), tetrabutylphosphonium (TBP), or an amino acid ester-and an anionic component-i.e., MTX. The biocompatibility, pharmacokinetics, tissue distribution, and antitumor efficacy of each MTX-based IL were investigated to determine its usefulness as a pharmaceutical. Oral administration to mice revealed that proline ethyl ester MTX (IL[ProEt][MTX]) had 4.6-fold higher oral bioavailability than MTX sodium, followed by aspartic diethyl ester MTX, IL[TBP][MTX], IL[Cho][MTX], and IL[TMA][MTX]. The peak plasma concentration, elimination half-life, area under the plasma concentration, mean absorption time, and body clearance of IL[ProEt][MTX] were significantly (p < 0.0001) higher by 1.7-, 6.2-, 4.6-, 2.5-, and 3.6-fold, respectively, than those of MTX sodium. MTX accumulation in the lungs, spleen, kidney, and gastrointestinal tract was also reduced by 5.6-, 1.8-, 1.5-, and 1.4-fold, respectively, indicating the IL formulations had lower systemic toxicity than free MTX. Mechanistic studies revealed that the IL[ProEt][MTX] solution formed spherical structures with an average size of 190 nm. This was probably responsible for its improved oral absorption performance in vivo. In vivo antitumor studies also demonstrated that IL[ProEt][MTX] suppressed tumor growth more than MTX sodium. These results suggest that MTX-based ILs provide a simple scalable approach to improving the oral bioavailability of poorly soluble MTX.


The Role of High-Potency Benzodiazepines in the Treatment of Panic Disorder.
Medication plays a central role in the treatment of panic disorder, with the goal of eliminating panic attacks and restoring normal function (i.e., achieving full remission). Four drug classes have similar efficacy (tricyclic antidepressants, selective serotonin reuptake inhibitors [SSRIs], benzodiazepines, and monoamine oxidase inhibitors). Nonetheless, benzodiazepines remain the most prescribed medication for panic disorder in the United States. The high-potency benzodiaze-pines alprazolam (available as immediate- and extended-release tablets) and clonazepam (available as tablets and orally disintegrating wafers) are the only benzodiazepines approved by the U.S. Food and Drug Administration for the treatment of panic disorder. High-potency benzodiaze-pines, with their proven efficacy in panic disorder exerted through control of the central nervous system excitability by a selective and potent enhancement of inhibitory gamma-aminobutyric acid-mediated neurotransmission, are also a safe and well-tolerated option for potentiation of rapid treatment response when initiating treatment with SSRIs. Judicious use of high-potency benzodiazepines followed by a cautious taper and discontinuation may optimize the benefits and minimize any potential risk associated with this class of drugs.


Improving central placement rates of peripherally inserted catheters.
Recent advances in catheter technology and in the knowledge base of professionals in the practice of intravenous therapy have had a significant impact on patient care and improved outcomes. As professionals learn new techniques and monitor outcomes, they appropriately begin to question some traditional beliefs. One of the items that we monitor in our continuous quality improvement program is the number of malpositioned peripherally inserted central catheters immediately after insertion. By reviewing the literature and discussing procedures with the hospital radiologist, a new method for advancing peripherally inserted central catheters for tip placement in the superior vena cava was evaluated, obtaining excellent results.


Matrix-assisted colloidosome reverse-phase layer-by-layer encapsulating biomolecules in hydrogel microcapsules with extremely high efficiency and retention stability.
The layer-by-layer (LbL) polyelectrolyte self-assembly encapsulation method has attracted much interest because of its versatility to use various polymers for capsule formation, ability to encapsulate different templates, and capability to control capsule permeability. Traditionally, the LbL method was performed in water as solvent and limited to poorly or non-water-soluble templates. Using the matrix-assisted LbL method, complex mixtures of water-soluble proteins or DNA could be encapsulated within agarose microbeads templates but leakage of biomolecules into the water phase during the LbL process results in low encapsulation efficiency. Recently, the reverse-phase LbL (RP-LbL) method was introduced to perform LbL and encapsulation of water-soluble templates in organic solvents, thus preventing the templates from dissolving and allowing high encapsulation efficiency. However, encapsulation of complex mixtures of biomolecules or other substances with quantitative encapsulation efficiency remained impossible. Here we present a new approach for encapsulation of biomolecules or complex mixtures thereof with almost 100% encapsulation efficiency. The ability of our method to achieve high encapsulation efficiency arises from the combination of two strategies. (1) Using microparticles as surface stabilizer to create stable biomolecule-loaded hydrogel microbeads, termed matrix-assisted colloidosome (MAC), that are able to disperse in oil and organic solvents. (2) Using the RP-LbL method to fabricate polymeric capsule "membranes", thereby preventing diffusion of the highly water-soluble biomolecules. Using an oil phase during emulsification and an organic solvent phase during encapsulation could completely prevent leakage of water-soluble biomolecules and almost 100% encapsulation efficiency is achieved. Microcapsules fabricated with our method retained nearly 100% of encapsulated proteins during a 7 day incubation period in water. The method was demonstrated on model proteins and may be extended to other biomolecules or mixtures. Our method is a valuable addition to the family of encapsulation techniques and can significantly contribute to the fields of bioreactors and bioanalytical microcapsules.


Nationwide survey on the current practice of ventricular tachycardia ablation.
We performed a nationwide survey on the current practice of ventricular tachycardia catheter ablation in Italy during the year 2016. Among 145 operators participating in the survey, 58 (40.0%) did not perform any ventricular tachycardia ablation in 2016. Among those performing ventricular tachycardia ablation, 9 operators (6.2%) performed only right ventricular endocardial catheter ablation, 52 (35.9%) performed endocardial catheter ablation both in the right and left ventricle (LV) and 26 (17.9%) performed both endocardial and epicardial LV catheter ablations. Seventy operators (89.7%) among the 78 performing LV and epicardial ablations treated patients with ischemic cardiomyopathy; ablations in the setting of other causes were less frequently performed. The following were considered as minimum requirements for ventricular tachycardia ablation: presence of a three-dimensional mapping system (120 operators, 82.8%), ICU in the hospital (118 operators, 81.4%), operator's training in high volume centers (93 operators, 64.1%). Twenty-eight operators (19.3%) performed catheter ablation in patients with electrical storm only after hemodynamic stabilization, 41 operators (28.3%) also during the acute phase and 9 operators (6.2%) never performed catheter ablation in electrical storm patients; the remaining 67 operators did not perform ventricular tachycardia ablation at all, or performed ablations only in the right ventricle. The present survey provides a snapshot of the current invasive treatment of ventricular tachycardia by catheter ablation. The procedure, especially in the setting of ischemic cardiomyopathy, is performed nationwide. Complex cases, including those with electrical storm, should be managed within a preestablished integrated network of regional referral centers able to transfer patients as soon as possible.


Scrutinizing assortative mating in birds.
It is often claimed that pair bonds preferentially form between individuals that resemble one another. Such assortative mating appears to be widespread throughout the animal kingdom. Yet it is unclear whether the apparent ubiquity of assortative mating arises primarily from mate choice ("like attracts like"), which can be constrained by same-sex competition for mates; from spatial or temporal separation; or from observer, reporting, publication, or search bias. Here, based on a conventional literature search, we find compelling meta-analytical evidence for size-assortative mating in birds (r = 0.178, 95% CI 0.142-0.215, 83 species, 35,591 pairs). However, our analyses reveal that this effect vanishes gradually with increased control of confounding factors. Specifically, the effect size decreased by 42% when we used previously unpublished data from nine long-term field studies, i.e., data free of reporting and publication bias (r = 0.103, 95% CI 0.074-0.132, eight species, 16,611 pairs). Moreover, in those data, assortative mating effectively disappeared when both partners were measured by independent observers or separately in space and time (mean r = 0.018, 95% CI -0.016-0.057). Likewise, we also found no evidence for assortative mating in a direct experimental test for mutual mate choice in captive populations of Zebra finches (r = -0.020, 95% CI -0.148-0.107, 1,414 pairs). These results highlight the importance of unpublished data in generating unbiased meta-analytical conclusions and suggest that the apparent ubiquity of assortative mating reported in the literature is overestimated and may not be driven by mate choice or mating competition for preferred mates.


Serum concentrations of adiponectin and characterization of adiponectin protein complexes in dogs.
To assess serum concentrations of adiponectin and characterize adiponectin protein complexes in healthy dogs. 11 healthy dogs. Sera collected from 10 dogs were evaluated via velocity sedimentation and ultracentrifugation, SDS-PAGE, western immunoblotting, and radioimmunoassay. Visceral adipose tissue (approx 90 g) was collected from the falciform ligament of a healthy dog undergoing elective ovariohysterectomy, and adiponectin gene expression was assessed via a real-time PCR procedure. Adiponectin gene expression was detected in visceral adipose tissue. Serum adiponectin concentrations ranged from 0.85 to 1.5 microg/mL (mean concentration, 1.22 microg/mL). In canine serum, adiponectin was present as a multimer, consisting of a low-molecular-weight complex (180 kd); as 3 (180-, 90-, and 60-kd) complexes under denaturing conditions; as 2 (90- and 60-kd) complexes under reducing conditions; and as a dimer, a monomer, and globular head region (60, 30, and 28 kd, respectively) under reducing-denaturing conditions. It is likely that adiponectin also circulates as a high-molecular-weight (360- to 540-kd) complex in canine serum, but resolution of this complex was not possible via SDS-PAGE. After exposure to identical experimental conditions, adiponectin protein complexes in canine serum were similar to those detected in human and rodent sera. Circulating adiponectin concentrations in canine serum were slightly lower than concentrations in human serum. Adiponectin gene expression was identified in canine visceral adipose tissue. Results suggest that adiponectin could be used as an early clinical marker for metabolic derangements, including obesity, insulin resistance, and diabetes mellitus in dogs.


Follicular fluid steroid and gonadotropic hormone levels and mitochondrial function from exosomes predict embryonic development.
Human follicular fluid (FF) is a complex biological fluid that contributes to the micro-environment of oocyte development. The aim of this study was to evaluate the role of steroid and gonadotropic hormones levels and mitochondrial function in embryo development during in vitro fertilization cycles. This was a cohort study of 138 women receiving IVF/ICSI, including 136 FF samples from 109 infertile women. FF steroid and gonadotropic hormones levels were tested by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and immunoassays. The mRNA expression levels of mitochondrial electron transport chain (ETC) complex genes from FF exosomes were detected by qPCR. Analysis of these individual FF concentrations revealed that LH and FSH concentrations were higher in follicles in which the oocyte developed into a top quality (TQ) blastocyst (LH: 9.44 ± 2.32mIU/ml, FSH: 9.32 ± 1.01mIU/ml) than those in which there was a failure of fertilization (LH: 5.30 ± 0.84mIU/ml, FSH: 6.91 ± 0.62mIU/ml). In contrast, follicular cortisone concentrations were lower for oocytes that resulted in a TQ blastocyst (12.20 ± 0.82mIU/ml). The receiver operating characteristic analysis showed that FF LH and FSH levels predicted TQ blastocyst with excellent AUC value of 0.711 and 0.747. Mitochondrial ETC complex I and III mRNA levels were increased in the FF exosomes of TQ blastocyst. Correlation analysis showed that mRNA levels of ETC complex I was positively correlated with LH and FSH levels in FF. The levels of FF steroid and gonadotropic hormones from single follicle can predetermine subsequent embryo development to some extent. Furthermore, impaired exosome mitochondrial dysfunction is a potiential event that causes hormone change in embryo development.


Postoperative arterial oxygen tension after peroperative PEEP-ventilation.
Forty otherwise healthy patients (29 women and 11 men), undergoing elective cholecystectomy, were randomly allocated to be ventilated during the operation either with a positive end-expiratory pressure of 1 kPa (10 cmH2O) (PEEP group) or with intermittent positive pressure ventilation without PEEP (control group). During the operation the mean arterial oxygen tension (PaO2) in the PEEP group increased from 14.6 to 16.5 kPa, while no changes occurred in the control group (13.5 and 13.6 kPa). On the first postoperative day, PaO2 decreased by 12% of the preoperative values in the PEEP group; the decrease was 20% in the control group. On the third postoperative day, the PaO2 in the control group was still 9% below the preoperative values, but on the fifth day, both groups had reached their preoperative PaO2 values. In the postoperative period, no statistically significant difference in PaO2 could be demonstrated between the groups. Determinations of the forced vital capacity and forced expiratory volume in the first second showed no difference between the groups pre- or postoperatively. The present study demonstrated no clinically relevant beneficial effect of peroperative PEEP ventilation on the postoperative arterial hypoxaemia after an upper abdominal laparotomy.


Ancient and conserved functional interplay between Bcl-2 family proteins in the mitochondrial pathway of apoptosis.
In metazoans, Bcl-2 family proteins are major regulators of mitochondrially mediated apoptosis; however, their evolution remains poorly understood. Here, we describe the molecular characterization of the four members of the Bcl-2 family in the most primitive metazoan, Trichoplax adhaerens All four trBcl-2 homologs are multimotif Bcl-2 group, with trBcl-2L1 and trBcl-2L2 being highly divergent antiapoptotic Bcl-2 members, whereas trBcl-2L3 and trBcl-2L4 are homologs of proapoptotic Bax and Bak, respectively. trBax expression permeabilizes the mitochondrial outer membrane, while trBak operates as a BH3-only sensitizer repressing antiapoptotic activities of trBcl-2L1 and trBcl-2L2. The crystal structure of a trBcl-2L2:trBak BH3 complex reveals that trBcl-2L2 uses the canonical Bcl-2 ligand binding groove to sequester trBak BH3, indicating that the structural basis for apoptosis control is conserved from T. adhaerens to mammals. Finally, we demonstrate that both trBax and trBak BH3 peptides bind selectively to human Bcl-2 homologs to sensitize cancer cells to chemotherapy treatment.


Microelectrode Array based Functional Testing of Pancreatic Islet Cells.
Electrophysiological techniques to characterize the functionality of islets of Langerhans have been limited to short-term, one-time recordings such as a patch clamp recording. We describe the use of microelectrode arrays (MEAs) to better understand the electrophysiology of dissociated islet cells in response to glucose in a real-time, non-invasive method over prolonged culture periods. Human islets were dissociated into singular cells and seeded onto MEA, which were cultured for up to 7 days. Immunofluorescent imaging revealed that several cellular subtypes of islets; β, δ, and γ cells were present after dissociation. At days 1, 3, 5, and 7 of culture, MEA recordings captured higher electrical activities of islet cells under 16.7 mM glucose (high glucose) than 1.1 mM glucose (low glucose) conditions. The fraction of the plateau phase (FOPP), which is the fraction of time with spiking activity recorded using the MEA, consistently showed distinguishably greater percentages of spiking activity with high glucose compared to the low glucose for all culture days. In parallel, glucose stimulated insulin secretion was measured revealing a diminished insulin response after day 3 of culture. Additionally, MEA spiking profiles were similar to the time course of insulin response when glucose concentration is switched from 1.1 to 16.7 mM. Our analyses suggest that extracellular recordings of dissociated islet cells using MEA is an effective approach to rapidly assess islet functionality, and could supplement standard assays such as glucose stimulate insulin response.


[Supraduodenal choledochoduodenal anastomosis in treating cholelithiasis].
In the recent years the number of patients with cholelithiasis has been increasing. The absolute majority of the patients are admitted to hospitals for urgent indications. Elderly patients with long histories, complications and concomitant diseases prevail. Not only the disease but complications as well must be exactly diagnosed. On the basis of the experience with 1298 operations on biliary ducts (178 of them subjected to choledochoduodenal anastomosis) the authors have determined the following indications for choledochoduodenal anastomosis: choledocholithiasis and dilated bile duct over 1,5 cm in diameter, cleavage of concrements in the bile duct, uncertainty of complete sanitation of the dilated duct and constricted terminal end of the choledochus with a concomitant pancreatitis. A one-row suture and the Yurash-Vinogradov method of choledochoduodenal anastomosis are preferred.


"The whole sky has broken down on me. I might die alone": A qualitative study on the lived experiences of COVID-19 positive frontline workers in Bangladesh.
Many countries, including Bangladesh, have conducted research on the mental health of frontline workers and their challenges in adjusting to their new workplaces. However, the authors are unaware of any studies on their real-life experiences as COVID-19-positive patients in Bangladesh. This study intends to investigate the lived experiences of Bangladeshi frontline workers who were isolated as a result of the COVID-19 infection and tested positive for the virus. We used a qualitative methodology and a semi-structured interview guide to conduct ten interviews between July 26 and August 12, 2020. The participants were recruited via a social media campaign and purposive sampling. All interviews were conducted via telephone and online and were transcribed and analyzed using Colaizzi's phenomenological method. The study does, however, identify four primary themes and 13 supporting themes, including (1) experience in a new working environment (subthemes: workload and adaptation, maintaining health protocol and social distance, and the fear of infection), (2) diagnosis (subthemes: the origin of infection, physiological problems, experiences at the diagnosis center), (3) recovery days (subthemes: earlier reactions, experiences in isolation, coping mechanisms), and (4) post-COVID-19 (subthemes: excitement, fear, and confusion; physiological problems; increased religiosity; and changes in philosophy). This study is important for healthcare policymakers because it helps them design healthcare management systems that take Bangladeshi society's social context into account. This study also recommends that long-term behavioral change programs be implemented by national policymakers to lessen societal stigma. At the same time, it suggests that the government should help lessen the barriers to health care services that persons with lower socioeconomic status confront.


On-demand release of Candida albicans biofilms from urinary catheters by mechanical surface deformation.
Candida albicans is a leading cause of catheter-associated urinary tract infections and elimination of these biofilm-based infections without antifungal agents would constitute a significant medical advance. A novel urinary catheter prototype that utilizes on-demand surface deformation is effective at eliminating bacterial biofilms and here the broader applicability of this prototype to remove fungal biofilms has been demonstrated. C. albicans biofilms were debonded from prototypes by selectively inflating four additional intralumens surrounding the main lumen of the catheters to provide the necessary surface strain to remove the adhered biofilm. Deformable catheters eliminated significantly more biofilm than the controls (>90% eliminated vs 10% control; p < 0.001). Mechanical testing revealed that fungal biofilms have an elastic modulus of 45 ± 6.7 kPa with a fracture energy of 0.4-2 J m-2. This study underscores the potential of mechanical disruption as a materials design strategy to combat fungal device-associated infections.


Cloning, characterization, and expression in Escherichia coli of a gene encoding Listeria seeligeri catalase, a bacterial enzyme highly homologous to mammalian catalases.
A gene coding for catalase (hydrogen-peroxide:hydrogen-peroxide oxidoreductase; EC 18.104.22.168) of the gram-positive bacterium Listeria seeligeri was cloned from a plasmid library of EcoRI-digested chromosomal DNA, with Escherichia coli DH5 alpha as a host. The recombinant catalase was expressed in E. coli to an enzymatic activity approximately 50 times that of the combined E. coli catalases. The nucleotide sequence was determined, and the deduced amino acid sequence revealed 43.2% amino acid sequence identity between bovine liver catalase and L. seeligeri catalase. Most of the amino acid residues which are involved in catalytic activity, the formation of the active center accession channel, and heme binding in bovine liver catalase were also present in L. seeligeri catalase at the corresponding positions. The recombinant protein contained 488 amino acid residues and had a calculated molecular weight of 55,869. The predicted isoelectric point was 5.0. Enzymatic and genetic analyses showed that there is most probably a single catalase of this type in L. seeligeri. A perfect 21-bp inverted repeat, which was highly homologous to previously reported binding sequences of the Fur (ferric uptake regulon) protein of E. coli, was detected next to the putative promoter region of the L. seeligeri catalase gene.


Scores of the Cleft Hearing, Appearance and Speech Questionnaire (CHASQ) in Swedish Participants With Cleft lip and/or Cleft Palate and a Control Population.
The primary aim of this study was to investigate whether there was any difference in scores of the Cleft Hearing, Appearance and Speech Questionnaire (CHASQ) between patients with cleft lip and/or cleft palate (CL/P) and a control population. The second aim was to compare CL/P and control population scores in this study with a British norm CL/P population. Single-site, cross-sectional study with an age-matched control population. Participants were recruited from a hospital, a school, and a sports club. They answered the CHASQ in the hospital or at home. Sixty-four participants with CL/P (7-19 years of age) and a control population of 56 participants without CL/P (9-20 years of age). CHASQ. There was no statistically significant difference in satisfaction with cleft-related features between the CL/P and the control population. Participants with CL/P were significantly more satisfied with non-cleft-related features than the control population. Cleft Hearing, Appearance and Speech Questionnaire scores were also similar to earlier established British normative data of a CL/P population. The results indicated that children and young people with CL/P were as satisfied with their appearance, hearing, and speech as children and young people without CL/P. Swedish CHASQ scores were also similar to British scores.


Estrogen treatment in vivo increases keratinocyte growth factor expression in the mammary gland.
Proliferation and differentiation of mammary epithelia are regulated by the combined action of systemic hormones and locally derived paracrine growth factors. Keratinocyte growth factor (KGF) is a potential candidate stromal factor that may participate in the hormonal control of stromal/epithelial interactions. In this study, we have examined the in vivo effect of 17beta-estradiol (E) treatment on KGF expression in mammary glands of peripubertal (5-week-old) and mature (11-week-old) mice. Mice received subcutaneous injections of hormone after which KGF mRNA levels were assayed by ribonuclease protection analysis of mammary gland RNA. E treatment caused a dose- and time-dependent increase in KGF mRNA levels in intact mice from both age groups. Neither 17alpha-estradiol nor progesterone injection affected KGF mRNA levels. Comparison of the relative expression of KGF in parenchyma-free fat pads and in intact glands demonstrated that the basal and E-dependent KGF mRNA levels did not require the presence of mammary epithelium. ELISA assay of KGF tissue content demonstrated that concomitantly with an up-regulation of mRNA, E treatment also increased KGF protein in mammary glands from peripubertal and mature mice. These data show that E treatment stimulates both KGF mRNA and protein expression in mammary stroma in vivo and raises the possibility that KGF has a role in E-regulated mammary gland development.


Testing group differences in brain functional connectivity: using correlations or partial correlations?
Resting-state functional magnetic resonance imaging allows one to study brain functional connectivity, partly motivated by evidence that patients with complex disorders, such as Alzheimer's disease, may have altered functional brain connectivity patterns as compared with healthy subjects. A functional connectivity network describes statistical associations of the neural activities among distinct and distant brain regions. Recently, there is a major interest in group-level functional network analysis; however, there is a relative lack of studies on statistical inference, such as significance testing for group comparisons. In particular, it is still debatable which statistic should be used to measure pairwise associations as the connectivity weights. Many functional connectivity studies have used either (full or marginal) correlations or partial correlations for pairwise associations. This article investigates the performance of using either correlations or partial correlations for testing group differences in brain connectivity, and how sparsity levels and topological structures of the connectivity would influence statistical power to detect group differences. Our results suggest that, in general, testing group differences in networks deviates from estimating networks. For example, high regularization in both covariance matrices and precision matrices may lead to higher statistical power; in particular, optimally selected regularization (e.g., by cross-validation or even at the true sparsity level) on the precision matrices with small estimation errors may have low power. Most importantly, and perhaps surprisingly, using either correlations or partial correlations may give very different testing results, depending on which of the covariance matrices and the precision matrices are sparse. Specifically, if the precision matrices are sparse, presumably and arguably a reasonable assumption, then using correlations often yields much higher powered and more stable testing results than using partial correlations; the conclusion is reversed if the covariance matrices, not the precision matrices, are sparse. These results may have useful implications to future studies on testing functional connectivity differences.


Pre-travel advice concerning vector-borne diseases received by travelers prior to visiting Cuzco, Peru.
Peru is an increasingly popular tourist destination that poses a risk to travelers due to endemic vector-borne diseases (VBDs). The objective of our study was to determine which factors are associated with receiving pre-travel advice (PTA) for VBDs among travelers visiting Cuzco, Peru. A cross-sectional secondary analysis based on data from a survey among travelers departing Cuzco at Alejandro Velazco Astete International Airport during the period January-March 2012 was conducted. From the 1819 travelers included in the original study, 1717 were included in secondary data analysis. Of these participants, 42.2% received PTA and 2.9% were informed about vector-borne diseases, including yellow fever (1.8%), malaria (1.6%) and dengue fever (0.1%). Receiving information on VBDs was associated with visiting areas endemic to yellow fever and dengue fever in Peru. The only disease travelers received specific recommendations for before visiting an endemic area for was yellow fever. Only 1 in 30 tourists received information on VBD prevention; few of those who traveled to an endemic area were warned about specific risks for infectious diseases prior to their trip. These important findings show that most tourists who travel to Peru do not receive PTA for the prevention of infectious and VBD, which can affect not only the travelers but their countries of origin as well.


Monoclonal antibody and F(ab')2 fragment delivery to tumor in patients with glioma: comparison of intracarotid and intravenous administration.
Non-i.v. delivery of radiolabeled monoclonal antibodies (MAbs) has been shown to increase tumor uptake and decrease dose to normal tissues. In this study, we have examined the potential advantage of intracarotid (i.c.) versus i.v. administration for the delivery of an intact MAb and a F(ab')2 fragment to tumor in patients with gliomas. Three patients received 10-50 mg of 81C6 IgG2b, a MAb reactive with the glioma-associated extracellular matrix antigen tenascin, and three received 5-20 mg of the F(ab')2 fragment of Mel-14, which is reactive with gliomas and melanomas. Paired-injection protocols, in which one-half of the MAb was labeled with 131I and administered by i.c. injection, and one-half was labeled with 125I and simultaneously administered by i.v. injection, were used. For both 81C6 IgG2b and Mel-14 (Fab')2, no differences in blood clearance half-times or urinary excretion rates of radioiodine were observed between i.c.- and i.v.-administered activity. Analysis of biopsy samples revealed i.c.:i.v. uptake ratios of 1.02 +/- 0.04, 0.95 +/- 0.03, and 1.03 +/- 0.05 for the accumulation of 81C6 IgG2b in temporalis muscle, normal brain, and glioma, respectively. Similarly, the i.c.:i.v. uptake ratios for Mel-14 F(ab')2 in these tissues were 0.98 +/- 0.04 (SD), 1.00 +/- 0.05, and 1.04 +/- 0.05. When the differences in percentage of injected dose/g uptake after i.c. and i.v. administration were compared, no statistically significant advantage for i.c. delivery was seen (P = 0.22-0.61). These data indicate that i.c. administration of MAb 81C6 IgG2b and Mel-14 F(ab')2 fragments offers no delivery advantage to offset the small but finite risk involved in cannulation and injection of the internal carotid artery.


De novo DNA methylation by Dnmt3a and Dnmt3b is dispensable for nuclear reprogramming of somatic cells to a pluripotent state.
Induced pluripotent stem cells (iPSCs) are generated from somatic cells by the transduction of defined transcription factors, and this process involves dynamic changes in DNA methylation. While the reprogramming of somatic cells is accompanied by demethylation of pluripotency genes, the functional importance of de novo DNA methylation has not been clarified. Here, using loss-of-function studies, we generated iPSCs from fibroblasts that were deficient in de novo DNA methylation mediated by Dnmt3a and Dnmt3b. These iPSCs reactivated pluripotency genes, underwent self-renewal, and showed restricted developmental potential that was rescued upon reintroduction of Dnmt3a and Dnmt3b. We conclude that de novo DNA methylation by Dnmt3a and Dnmt3b is dispensable for nuclear reprogramming of somatic cells.


Estimating HIV prevalence and projecting AIDS incidence in the United States: a model that accounts for therapy and changes in the surveillance definition of AIDS.
The AIDS incubation distribution is changing in calendar time because of treatment and changes in the surveillance definition of AIDS. To obtain reliable estimates of HIV prevalence and projections of AIDS incidence in the 1990s using the method of backcalculation, we constructed an appropriate incubation distribution for each calendar date of infection. We parameterized the impact of treatment on the incubation distribution by specifying the relative hazard for AIDS in treated versus untreated people as a function of duration of HIV infection. To account for trends in the incubation distribution, we modelled the prevalence of treatment, the distribution of treatment onset times, and the impact of the revision of the AIDS surveillance definition in 1987. We selected and evaluated backcalculation models based on consistency with external information. We defined a 'plausible range' of estimates that took into account uncertainty about the natural incubation distribution and treatment efficacy, as well as bootstrap assessment of stochastic error. Using these methods, we projected that national United States AIDS incidence will plateau during 1991-1994 at over 50,000 cases per year. Projections exhibited substantial systematic uncertainty, and we calculated a plausible range for AIDS incidence in 1994 of 42,300 to 70,700 cases. An estimated 628,000 to 988,000 cumulative HIV infections occurred as of 1 January 1991. After accounting for AIDS mortality, we estimated that 484,000 to 844,000 people were living with HIV infection on 1 January 1991. Favourable trends in HIV incidence appeared in gay men and intravenous drug users. Plausible ranges for our estimates overlapped with those from a 'stage model' approach to incorporating treatment effects in backcalculations. Our approach, however, tended to yield smaller estimates of epidemic size, mainly because the parameters used with the stage model implied that more treatment was in use and that treatment was more effective than in our model.


Genetic structure in the Mediterranean seagrass Posidonia oceanica: disentangling past vicariance events from contemporary patterns of gene flow.
The Mediterranean Sea is a two-basin system, with the boundary zone restricted to the Strait of Sicily and the narrow Strait of Messina. Two main population groups are recognized in the Mediterranean endemic seagrass Posidonia oceanica, corresponding to the Western and the Eastern basins. To address the nature of the East-West cleavage in P. oceanica, the main aims of this study were: (i) to define the genetic structure within the potential contact zone (i.e. the Strait of Sicily) and clarify the extent of gene flow between the two population groups, and (ii) to investigate the role of present water circulation patterns vs. past evolutionary events on the observed genetic pattern. To achieve these goals, we utilized SSR markers and we simulated, with respect to current regime, the possible present-day dispersal pattern of Posidonia floating fruits using 28-day numerical Lagrangian trajectories. The results obtained confirm the presence of the two main population groups, without any indices of reproductive isolation, with the break zone located at the level of the Southern tip of Calabria. The populations in the Strait of Sicily showed higher affinity with Western than with Eastern populations. This pattern of genetic structure probably reflects historical avenues of recolonization from relict glacial areas and past vicariance events, but seems to persist as a result of the low connectivity among populations via marine currents, as suggested by our dispersal simulation analysis.


Metabolic aspects of bariatric surgery.
Insulin resistance is a nearly universal finding in morbid obesity. It may be compensated and latent or uncompensated with single or multiple clinical abnormalities. Although lifestyle interventions and medical measures alone may control most metabolic problems in the short term, the ultimate benefits of such an approach are usually limited by the complexity of available therapeutic regimens and the difficulty of maintaining full patient compliance. Many studies now document that bariatric surgery can effectively and safely control these complications in the short term and long term or even prevent their occurrence. Further investigations are needed to understand better the mechanisms involved and to define more clearly the appropriate indications and contraindications of the treatments proposed.


Molecular control of the cytosolic aconitase/IRP1 switch by extramitochondrial frataxin.
The inability to produce normal levels of the mitochondrial protein frataxin causes the hereditary degenerative disorder Friedreich's Ataxia (FRDA), a syndrome characterized by progressive gait instability, cardiomyopathy and high incidence of diabetes. Frataxin is an iron-binding protein involved in the biogenesis of iron-sulfur clusters (ISC), prosthetic groups allowing essential cellular functions such as oxidative phosphorylation, enzyme catalysis and gene regulation. Although several evidence suggest that frataxin acts as an iron-chaperone within the mitochondrial compartment, we have recently demonstrated the existence of a functional extramitochondrial pool of mature frataxin in various human cell types. Here, we show that a similar proteolytic process generates both mature mitochondrial and extramitochondrial frataxin. To address the physiological function of human extramitochondrial frataxin, we searched for ISC-dependent interaction partners. We demonstrate that the extramitochondrial form of frataxin directly interacts with cytosolic aconitase/iron regulatory protein-1 (IRP1), a bifunctional protein alternating between an enzymatic and a RNA-binding function through the 'iron-sulfur switch' mechanism. Importantly, we found that the cytosolic aconitase defect and consequent IRP1 activation occurring in FRDA cells are reversed by the action of extramitochondrial frataxin. These results provide new insight into the control of cytosolic aconitase/IRP1 switch and expand current knowledge about the molecular pathogenesis of FRDA.


Measuring time-varying information flow in scalp EEG signals: orthogonalized partial directed coherence.
This study aimed to develop a time-frequency method for measuring directional interactions over time and frequency from scalp-recorded electroencephalographic (EEG) signals in a way that is less affected by volume conduction and amplitude scaling. We modified the time-varying generalized partial directed coherence (tv-gPDC) method, by orthogonalization of the strictly causal multivariate autoregressive model coefficients, to minimize the effect of mutual sources. The novel measure, generalized orthogonalized PDC (gOPDC), was tested first using two simulated models with feature dimensions relevant to EEG activities. We then used the method for assessing event-related directional information flow from flash-evoked responses in neonatal EEG. For testing statistical significance of the findings, we followed a thresholding procedure driven by baseline periods in the same EEG activity. The results suggest that the gOPDC method 1) is able to remove common components akin to volume conduction effect in the scalp EEG, 2) handles the potential challenge with different amplitude scaling within multichannel signals, and 3) can detect directed information flow within a subsecond time scale in nonstationary multichannel EEG datasets. This method holds promise for estimating directed interactions between scalp EEG channels that are commonly affected by the confounding impact of mutual cortical sources.


Distressing near-death experiences.
Most reported near-death experiences include profound feelings of peace, joy, and cosmic unity. Less familiar are the reports following close brushes with death of experiences that are partially or entirely unpleasant, frightening, or frankly hellish. While little is known about the antecedents or aftereffects of these distressing experiences, there appear to be three distinct types, involving (1) phenomenology similar to peaceful near-death experiences but interpreted as unpleasant, (2) a sense of nonexistence or eternal void, or (3) graphic hellish landscapes and entities. While the first type may eventually convert to a typical peaceful experience, the relationship of all three types to prototypical near-death experiences merits further study. The effect of the distressing experience in the lives of individuals deserves exploration, as the psychological impact may be profound and long-lasting.


A clinically meaningful theory of outcome measures in rehabilitation medicine.
Comparative effectiveness research in rehabilitation medicine requires the development and validation of clinically meaningful and scientifically rigorous measurements of patient states and theories that explain and predict outcomes of intervention. Patient traits are latent (unobservable) variables that can be measured only by inference from observations of surrogate manifest (observable) variables. In the behavioral sciences, latent variables are analogous to intensive physical variables such as temperature and manifest variables are analogous to extensive physical variables such as distance. Although only one variable at a time can be measured, the variable can have a multidimensional structure that must be understood in order to explain disagreements among different measures of the same variable. The use of Rasch theory to measure latent trait variables can be illustrated with a balance scale metaphor that has randomly added variability in the weights of the objects being measured. Knowledge of the distribution of the randomly added variability provides the theoretical structure for estimating measures from ordinal observation scores (e.g., performance measures or rating scales) using statistical inference. In rehabilitation medicine, the latent variable of primary interest is the patient's functional ability. Functional ability can be estimated from observations of surrogate performance measures (e.g., speed and accuracy) or self-report of the difficulty the patient experiences performing specific activities. A theoretical framework borrowed from project management, called the Activity Breakdown Structure (ABS), guides the choice of activities for assessment, based on the patient's value judgments, to make the observations clinically meaningful. In the case of low vision, the functional ability measure estimated from Rasch analysis of activity difficulty ratings was discovered to be a two-dimensional variable. The two visual function dimensions are independent of physical limitations and psychological state. To explain outcome measures (latent variable estimated from difficulty ratings), a theory must be developed that explicitly defines how latent variables are related to the observed manifest variables and to each other. The latent variables are categorized as primary variables, which in the case of low vision are the two visual function dimensions, and as effect modifiers, which in the case of low vision are other physical and psychological latent traits of the patients that can influence the outcome measures. Interventions give rise to latent intervention effect variables that can alter the latent primary variables or independently affect the outcome measures. The latent effect modifier variables, in turn, can alter the latent intervention effect variables. Once developed and validated, a theory of this form will predict the rehabilitation potential of individual patients, i.e., the probability of obtaining criterion outcome measures given the observed state of the patient and the choice of interventions.


Evaluation of transrectal ultrasound in the early detection of prostate cancer.
To determine the ability of transrectal ultrasound to detect early localized prostate cancer, unsuspected (nonpalpable) cancer in the contralateral lobe of patients undergoing radical prostatectomy for clinically localized disease was evaluated. A total of 59 patients with palpable prostate cancer clinically confined to 1 lobe underwent transrectal ultrasound before radical prostatectomy and step-sectioning of the radical prostatectomy specimen. Transrectal ultrasound was performed with 5 or 7 MHz. real-time transrectal units. Pathological findings in these 59 cases revealed no tumor in the contralateral lobe in 34 (58%) and the presence of unsuspected tumor in 25 (42%). Transrectal ultrasound detected 13 of 25 unsuspected cancers for a sensitivity of 52%. Of 34 patients with no contralateral lobe lesion transrectal ultrasound was correct in 23 for a specificity of 68%. The positive and negative predictive values for transrectal ultrasound in this study group were 54 and 66%, respectively. There was no significant difference in the pathological size of the clinically suspected and clinically unsuspected cancers as measured by average largest dimension, and transrectal ultrasound sensitivity did not correlate with the size of the cancer. Based on careful sonopathological analysis, transrectal ultrasound may not be a good method to detect clinically unsuspected prostate cancer and the false positive rate would appear to be high.


Critical evaluation of root causes of the reduced compactability after roll compaction/dry granulation.
The influence of lubrication and particle size on the reduced compactability after dry granulation was investigated. Powder cellulose, lactose, magnesium carbonate, and two types of microcrystalline cellulose were roll compacted, granulated, and sieved into particle fractions. Particle fractions were compressed into tablets using internal and external lubrication. Internal lubrication resulted in an overlubrication of the granule material compared with the powder material. This resulted in extraordinary high reduction of compactability after dry granulation for lubricant-sensitive materials. The granule size can cause differences in strength, whereby the degree of this effect was material dependent. The loss in strength with increasing compaction force was comparable for different particles sizes of one material, suggesting a change in material properties independently of the size. Granule hardening could be one reason as for higher compaction forces the integrity of the granule structure survived the compression step. The results demonstrated that granule lubrication mainly influence the degree of the reduced compactability after dry granulation and must be considered for the evaluation of mechanism for this phenomenon. Hardening of the material as well as size enlargement will cause the loss in strength after recompression, but the influence of both depends strongly on the material.


Increase in furfural tolerance in ethanologenic Escherichia coli LY180 by plasmid-based expression of thyA.
Furfural is an inhibitory side product formed during the depolymerization of hemicellulose by mineral acids. Genomic libraries from three different bacteria (Bacillus subtilis YB886, Escherichia coli NC3, and Zymomonas mobilis CP4) were screened for genes that conferred furfural resistance on plates. Beneficial plasmids containing the thyA gene (coding for thymidylate synthase) were recovered from all three organisms. Expression of this key gene in the de novo pathway for dTMP biosynthesis improved furfural resistance on plates and during fermentation. A similar benefit was observed by supplementation with thymine, thymidine, or the combination of tetrahydrofolate and serine (precursors for 5,10-methylenetetrahydrofolate, the methyl donor for ThyA). Supplementation with deoxyuridine provided a small benefit, and deoxyribose was of no benefit for furfural tolerance. A combination of thymidine and plasmid expression of thyA was no more effective than either alone. Together, these results demonstrate that furfural tolerance is increased by approaches that increase the supply of pyrimidine deoxyribonucleotides. However, ThyA activity was not directly affected by the addition of furfural. Furfural has been previously shown to damage DNA in E. coli and to activate a cellular response to oxidative damage in yeast. The added burden of repairing furfural-damaged DNA in E. coli would be expected to increase the cellular requirement for dTMP. Increased expression of thyA (E. coli, B. subtilis, or Z. mobilis), supplementation of cultures with thymidine, and supplementation with precursors for 5,10-methylenetetrahydrofolate (methyl donor) are each proposed to increase furfural tolerance by increasing the availability of dTMP for DNA repair.


Phosphorylation of eukaryotic initiation factor-2{alpha} promotes the extracellular survival of obligate intracellular parasite Toxoplasma gondii.
While seeking a new host cell, obligate intracellular parasites, such as the protozoan Toxoplasma gondii, must be able to endure the stress of an extracellular environment. The mechanisms Toxoplasma use to remain viable while deprived of a host cell are not understood. We have previously shown that phosphorylation of Toxoplasma eukaryotic initiation factor-2α (TgIF2α) is a conserved response to stress. Here we report the characterization of Toxoplasma harboring a point mutation (S71A) in TgIF2α that prevents phosphorylation. Results show that TgIF2α phosphorylation is critical for parasite viability because the TgIF2α-S71A mutants are ill-equipped to cope with life outside the host cell. The TgIF2α-S71A mutants also showed a significant delay in producing acute toxoplasmosis in vivo. We conclude that the phosphorylation of TgIF2α plays a crucial role during the lytic cycle by ameliorating the stress of the extracellular environment while the parasite searches for a new host cell.