Cardiac but not pulmonary receptors mediate depressor response to IV phenyl biguanide in conscious rabbits.
Phenyl biguanide (PBG) stimulates pulmonary and cardiac receptors in the cat and rabbit. Previous reports have suggested that pulmonary receptors mediate the reflex respiratory responses and cardiac receptors mediate the reflex hypotension and bradycardia. Using selective denervation of the lung (LDX) and intrapericardial procaine to block cardiac nerves (CDX), we investigated the specific role of pulmonary and cardiac receptors in the reflex response to PBG infusion (60 micrograms/kg iv) in the conscious rabbit. Breathing frequency, arterial pressure, heart rate, and renal sympathetic nerve activity (RSNA) were recorded before and after LDX and CDX. Before LDX and CDX, PBG infusion produced tachypnea, hypotension, and bradycardia. This was accompanied by a withdrawal of RSNA to a level not different from that evoked by ganglionic blockade. These responses were preserved after LDX but were abolished after CDX with intrapericardial procaine. Intrapericardial infusion of PBG produced no response. These results indicate that in conscious rabbits both the respiratory and cardiovascular responses to PBG infusion are mediated by cardiac receptors not accessible from the epicardial surface. Furthermore, the reflex hypotension is mediated largely by withdrawal of sympathetic nerve activity resulting in a decreased peripheral resistance.


Evaluating the level of injury in English professional football using a risk based assessment process.
To show how epidemiological data can be presented and analysed in frequency based and risk based formats and how risk based information can simplify management decisions on injury prevention strategies in professional football. The club physiotherapists at four English professional football clubs prospectively recorded players' injuries over the period November 1994 to May 1997. The nature, location, and mechanism of each injury and the specific numbers of days that players were unavailable to train or play as a result of injuries were recorded. The rates of injury were evaluated on a risk matrix using the number of days and the estimated costs of absence as measures of injury consequences. There was a significant difference in the time lost through injury as a function of injury severity (p<0.001). Slight and minor injuries accounted for 51% of all injuries but represented only 17% of the risk from injury, whereas major injuries accounted for only 12% of the number of injuries and 47% of the risk. Player to player contact injuries accounted for 39% of the risk of injury, and football specific activities accounted for 47% of the risk. The risks of acute injury in professional football were three orders of magnitude greater than those in the construction, manufacturing, and service sectors of industry. The risks associated with minor, moderate, and major acute injuries and osteoarthritis in lower limb joints of professional footballers were found to be unacceptable when evaluated against work based risk criteria used by the Health and Safety Executive. All stakeholders within professional football were shown to have an important contribution to make in reducing the overall level of risk to players through the provision of risk prevention strategies.


A case of echinostomiasis with ulcerative lesions in the duodenum.
Echinostomiasis is an endemic intestinal trematodiasis of humans in Korea. We observed a human case of Echinostoma hortense infection who had ulcerations on the duodenal mucosa. A 55-year old man living in Hamyang-gun, Kyongnam, complained of epigastric pain with hematemesis in April 1994. Endoscopy revealed lesions of early gastric cancer and duodenal ulcerations. A penetrating parasite into the duodenal mucosa was picked out, and identified as E. hortense. As the patient was treated with praziquantel 10 mg/kg single dose, 3 more E. hortense and 7 Metagonimus worms were recovered. This case demonstrates that echinostomiasis causes gross ulcerations in the duodenum.


Cloning and nucleotide sequence of the isoamylase gene from a strain of Pseudomonas sp.
A strain of Pseudomonas sp., SMP1, isolated from a soil sample collected in the Monterotondo area (Rome), secreted isoamylase activity into the culture medium. The enzyme was purified and optimal reaction and stability conditions were determined by varying pH and temperature. The chemico-physical properties of the enzyme were similar to those of the isoamylase purified in Japan more than 20 years ago from 'Pseudomonas amyloderamosa' strain SB15. A genomic library of SMP1 was prepared in Escherichia coli using pUC12 as vector. Two isoamylase-producing colonies were identified out of 6300 screened. The hybrid plasmids isolated from the two clones showed common restriction patterns. The chromosomal portion of one of these plasmids (pSM257) was completely sequenced. Comparison between the deduced amino acid sequence of the isoamylase and the published sequences of other amylolytic enzymes showed the presence of conserved domains.


Management services organizations: providing economies of scale.
Effective information management and other shared costs can reduce agencies' operating expenses as well as make new products available while providing opportunities to gain a competitive advantage. By taking a close look at ways of controlling documentation, such as the use of a management services organization for information system/computer-based management, agencies can prepare for the challenges of the next century.


Scapular fractures: a common diagnostic pitfall.
Scapular fractures are one of the most difficult fractures to diagnose on radiographs. Detection can be challenging because of the obscuration by the overlying structures or incomplete imaging due to difficult patient collaboration. Familiarity with imaging characteristics of these abnormalities will allow radiologists to better diagnose and characterize scapular fractures. Three-dimensional computed tomographic scans are considered the gold standard for scapular diagnoses. Treatment strategies differ depending on the type of scapular fractures, but the site and degree of displacement will determine whether surgical intervention should be considered. Complications can occur in fractures that are undiagnosed or improperly evaluated. The purpose of this article is to describe imaging features of traumatic scapular injury, and discuss the role of diagnostic imaging in clinical decision making after shoulder trauma.


Virulent avian influenza A viruses: their effect on avian lymphocytes and macrophages in vivo and in vitro.
To investigate the pathogenesis of virulent avian influenza A viruses, the effect of A/turkey/Ont/7732/66 (H5N9) (Ty/Ont), A/tern/South Africa/1961 (H5N3) (Tern/S.A.) and A/chicken/Pennsylvania/1370/83 (H5N2) (Ck/Penn) on avian lymphoid cell populations was examined in vivo. Previous studies have shown that infection of chickens with Ty/Ont resulted in the extensive destruction of lymphoid tissues. In this study, other virulent avian H5 influenza viruses, Tern/S.A. or Ck/Penn, had little or no effect on lymphoid tissues of infected chickens. Therefore the effect of Ty/Ont on lymphoid tissue is a specific activity of this virus only and not of other virulent avian H5 influenza strains. To examine the role of viral replication in the destruction of lymphocytes, in vitro cultures of avian macrophages and lymphocytes were inoculated with Ty/Ont. Macrophages supported the synthesis of viral proteins whereas lymphocytes produced small, but detectable amounts of viral protein; however, infectious virus was not produced by either cell type. Furthermore inoculation of chicken spleen cells with Ty/Ont in vivo and in vitro had a profound effect on the proliferative response of lymphocytes to concanavalin A. These results suggest that Ty/Ont infects macrophages as well as lymphocytes in the chicken, and the effects of the virus on both cell types may well contribute to lymphoid necrosis.


[Tobacco and cannabis consumption : toxicity and management of quitting].
The use of cannabis is most often associated with that of tobacco. While the addictive power of cannabis is low, that of tobacco is high. The regular use of both tobacco and cannabis is at the origin of the increase in dependence on each substance, which makes stopping this double consumption often difficult. This combined use can cause somatic and mental complications. Practitioners should identify this dual use, advise stopping and offer their help after a careful evaluation of both consumptions and their causes. The management includes psychotherapeutic support; If the efficacy of medications to aid stopping cannabis has not been proven to aid in stopping cannabis, nicotine replacement therapy can limit withdrawal syndrome, craving and improve patients' adherence to monitoring.The mobilization of practitioners in the management of patients and the development of effective cannabis cessation pharmacotherapies are essential.


Hypermutability in environmental Pseudomonas aeruginosa and in populations causing pulmonary infection in individuals with cystic fibrosis.
Pseudomonas aeruginosa is the pathogen most commonly associated with morbidity and mortality in cystic fibrosis (CF) patients. The host-pathogen interactions responsible for progressive CF lung diseases are complex. However, there is growing interest in the role of hypermutable P. aeruginosa (that is, those strains with an increased mutation frequency due to mutations in mismatch repair and error prevention genes), in terms of both bacterial adaptation and antimicrobial resistance. The prevalence of hypermutable P. aeruginosa in chronic CF infection has been established, and at 37 % is surprisingly high. To the authors' knowledge, there are no reports of prevalence during the early stages of infection, in environmental pseudomonas, which are believed to be the primary source of infection, and in epidemic strains, which have emerged as a major challenge. The aim of this study was to establish the prevalence of hypermutable P. aeruginosa in these pseudomonas populations. The hypothesis was that hypermutability would be rare in early and in environmental P. aeruginosa but in contrast would explain the relatively recent emergence of epidemic strains. It was found that 10/100 (10 %) of early isolates were strong or weak mutators, suggesting that the CF lung is not the only factor influencing the existence of mutators in this group of patients. Two weak mutators (6 %) were found in 32 environmental isolates. Only two of 15 (13 %) epidemic P. aeruginosa strains were hypermutable, and although closer analysis revealed this issue to be complex, on the whole the data suggested that the atypical characteristics of these highly transmissible strains cannot solely be explained by this phenomenon. The higher than predicted prevalence of mutators in early infection, and in environmental isolates, reinforces the importance of early and aggressive treatment for P. aeruginosa infection in CF.


Protein O-mannosyltransferase activities in lymphoblasts from patients with alpha-dystroglycanopathies.
Defects in O-mannosylation of alpha-dystroglycan cause some forms of congenital muscular dystrophy (CMD), the so-called alpha-dystroglycanopathies. Six genes are responsible for these diseases with overlapping phenotypes. We investigated the usefulness of a biochemical approach for the diagnosis and investigation of the alpha-dystroglycanopathies using immortalized lymphoblasts prepared from genetically diagnosed and undiagnosed CMD patients and from control subjects. We measured the activities of protein O-mannose beta1,2-N-acetylglucosaminyltransferase 1 (POMGnT1) and protein O-mannosyltransferase (POMT). Lymphoblasts from patients harbouring known mutations in either POMGNT1 or POMT1 showed a marked decrease in POMGnT1 or POMT activity, respectively, compared to controls. Furthermore, we identified pathogenic mutations in POMGNT1, POMT1 or POMT2 in six previously genetically uncharacterised patients who had very low enzyme activity. In conclusion, the lymphoblast-based enzymatic assay is a sensitive and useful method (i) to select patients harbouring POMGNT1, POMT1 or POMT2 mutations; (ii) to assess the pathogenicity of new or already described mutations.


Differential TERT promoter methylation and response to 5-aza-2'-deoxycytidine in acute myeloid leukemia cell lines: TERT expression, telomerase activity, telomere length, and cell death.
The catalytic subunit of human telomerase (TERT) is highly expressed in cancer cells, and correlates with complex cytogenetics and disease severity in acute myeloid leukemia (AML). The TERT promoter is situated within a large CpG island, suggesting that expression is methylation-sensitive. Studies suggest a correlation between hypermethylation and TERT overexpression. We investigated the relationship between TERT promoter methylation and expression and telomerase activity in human leukemia and lymphoma cell lines. DAC-induced demethylation and cell death were observed in all three cell lines, as well as telomere shortening in HL-60 cells. DAC treatment reduced TERT expression and telomerase activity in OCI/AML3 and HL-60 cells, but not in U937 cells. Control U937 cells expressed lower levels of TERT mRNA, carried a highly methylated TERT core promoter, and proved more resistant to DAC-induced repression of TERT expression and cell death. AML patients had significantly lower methylation levels at several CpGs than "well elderly" individuals. This study, the first to investigate the relationship between TERT methylation and telomerase activity in leukemia cells, demonstrated a differential methylation pattern and response to DAC in three AML cell lines. We suggest that, although DAC treatment reduces TERT expression and telomerase activity, this is unlikely to occur via direct demethylation of the TERT promoter. However, further investigations on the regions spanning CpGs 7-12 and 14-16 may reveal valuable information regarding transcriptional regulation of TERT.


Effects of atrial natriuretic factor on cyclic guanosine monophosphate and cyclic adenosine monophosphate accumulation in microdissected nephron segments from rats.
Atrial natriuretic factor (ANF) (1 microM) markedly increased cyclic guanosine monophosphate (cGMP) content in microdissected glomeruli (35-fold) and in microdissected inner medullary collecting ducts (IMCD) (20-fold). ANF caused little or no increase in cGMP content in other nephron segments. The threshold concentration for increased cGMP accumulation by ANF was 0.1-1 nM in IMCD, which is in the range reported for rat plasma. Sodium nitroprusside (1 mM), which selectively stimulates soluble guanylate cyclase, increased cGMP content in glomeruli but not in IMCD. ANF did not alter cAMP accumulation in the absence or presence of vasopressin (AVP) or parathyroid hormone (PTH) in outer and inner medullary tubule suspensions, or in microdissected proximal convoluted tubules (PCT), medullary thick ascending limbs (MAL) or IMCD. These data are compatible with the hypothesis that cGMP is a second messenger for a physiologic action of ANF in the inner medullary collecting duct. ANF apparently activates membrane-bound guanylate cyclase in this segment.


[Can bariatric surgery cure metabolic syndrome?].
To evaluate the impact of bariatric surgery on the metabolic syndrome (MS) and what are the criteria that contribute the most for its exclusion after surgery. The progress of leucometry was also analyzed. 47 obese women with MS were evaluated. All patients were operated with the Roux-en-Y vertical gastric bypass technique, with the insertion of a contention ring on the gastro-jejune anastomosis (Fobi-Capella). Patients were evaluated before and on the first year post-surgery. Fasting glucose presented a relevant decrease at 3 months after surgery. After 12 months, all 20 patients who had DM2 or altered fasting glucose presented normal levels of fasting glucose and glicated hemoglobin, and none of them was using any anti-diabetic drug. Triglycerides levels were reduced by 49.2%, whereas HDL-cholesterol increased by 27.2%. Systolic and diastolic blood pressures were reduced by an average of 28.7 and 20.8 mmHg, respectively. Leucocytes counting fell from 7671/microL to 6156/microL. Fasting glucose, triglycerides, DBP, SBP and HDL-cholesterol were the variables that contributed most for the reduction of MS. At the end of the first year, elimination of MS occurred in 80.9% of the patients. Bariatric surgery reduces resistance to insulin and consequently the cardiovascular risk factors.


Torsade de Pointes Due to Hypokalemia and Hypomagnesemia.
This scenario was developed to educate emergency medicine (EM) interns but can be used to educate medical students and junior residents. Torsade de Pointes (TdP) is a rare but potentially fatal arrythmia if not quickly diagnosed and properly treated. TdP is defined as a polymorphic ventricular tachycardia (VT) characterized by an oscillatory change in amplitude around an isoelectric line that is associated with a QTc prolongation on the electrocardiogram (ECG).1 It has been well described to predispose to ventricular fibrillation and arrhythmic death. QTc prolongation can be congenital or acquired. Between 1 in 2000 to 20,000 have the genetic mutation for QTc prolongation.1 Acquired QTc is most commonly drug related leading to electrolyte abnormalities. 2 Around 28% of cases of TdP are associated with hypokalemia and hypomagnesemia.2 Several European centers estimate 0.8 to 1.2 per million people per year are drug induced.1 Patients with TdP most commonly presents with syncope, palpitations, and dizziness.2 While 50% are asymptomatic, up to 10% of patients will present in cardiac arrest.1 It is imperative for EM physicians to be able to recognize TdP as it can quickly decompensate into a ventricular fibrillation and sudden death. These patients require management of electrolyte abnormalities, ventricular dysrhythmias, and cardiac death.2 This simulation case will demonstrate treatment strategies for TdP with electrolyte repletion, antiarrhythmics, and defibrillation. By the end of this simulation session, learners will be able to: 1) formulate appropriate work-up for altered mental status (AMS) 2) recognize hypokalemia and associated findings on ECG 3) address hypomagnesemia in a setting to hypokalemia 4) manage pulseless VT by following advanced cardiac life support (ACLS) 5) recognize and address TdP 6) provide care after return of spontaneous circulation (ROSC) 7) consult intensivist and admit to intensive care unit (ICU). This session was conducted using high-fidelity simulation, which was immediately followed by an in-depth debriefing session. Each session had three EM first-year residents and six observers. There was one simulation instructor running the session and one simulation technician who acted as a nurse. After the simulation and debriefing session was complete, an online survey was sent via surveymonkey.com to all the participants. The survey collected responses to the following questions: (1) was the case believable? (2) did the case have the right amount of complexity? (3) did the case help improve medical knowledge and patient care? (4) did the simulation environment gave a real-life experience? (5) did the debriefing session after simulation help improve knowledge? A Likert scale was used to collect the responses. This case was performed once a year for 2 years in a row. There was a total of 19 respondents from both years. One hundred percent of them either agreed or strongly agreed that the case was beneficial in learning and in improving medical knowledge and patient care. All of them found the post-session debrief to be very helpful. Two of them felt neutral about the case being realistic. This high-fidelity simulation was a realistic way of educating learners on how to manage hypokalemia and hypomagnesemia leading to TdP. Cost-effectiveness varies depending on what is available at individual simulation laboratories. Learners are forced to start with a broad differential for the patient who presents with AMS. As they manage the case, the patient quickly decompensates into a fatal arrhythmia due to electrolyte abnormalities. Learners enforced their knowledge on leading ACLS, intubation skills, and treating TdP with electrical conversion and electrolyte repletion. Hypokalemia, hypomagnesemia, torsades de pointes, altered mental status, medical simulation.


Using the ratio of means as the effect size measure in combining results of microarray experiments.
Development of efficient analytic methodologies for combining microarray results is a major challenge in gene expression analysis. The widely used effect size models are thought to provide an efficient modeling framework for this purpose, where the measures of association for each study and each gene are combined, weighted by the standard errors. A significant disadvantage of this strategy is that the quality of different data sets may be highly variable, but this information is usually neglected during the integration. Moreover, it is widely known that the estimated standard deviations are probably unstable in the commonly used effect size measures (such as standardized mean difference) when sample sizes in each group are small. We propose a re-parameterization of the traditional mean difference based effect measure by using the log ratio of means as an effect size measure for each gene in each study. The estimated effect sizes for all studies were then combined under two modeling frameworks: the quality-unweighted random effects models and the quality-weighted random effects models. We defined the quality measure as a function of the detection p-value, which indicates whether a transcript is reliably detected or not on the Affymetrix gene chip. The new effect size measure is evaluated and compared under the quality-weighted and quality-unweighted data integration frameworks using simulated data sets, and also in several data sets of prostate cancer patients and controls. We focus on identifying differentially expressed biomarkers for prediction of cancer outcomes. Our results show that the proposed effect size measure (log ratio of means) has better power to identify differentially expressed genes, and that the detected genes have better performance in predicting cancer outcomes than the commonly used effect size measure, the standardized mean difference (SMD), under both quality-weighted and quality-unweighted data integration frameworks. The new effect size measure and the quality-weighted microarray data integration framework provide efficient ways to combine microarray results.


Parasites of two native fishes in adjacent Adirondack lakes.
This survey of parasites in 2 adjacent lakes is the first of its kind in the Adirondack Park of New York State. Wolf Lake is designated as a heritage lake whereas nearby Deer Lake is limnologically similar but has at least 5 introduced fish species. Both lakes have 2 native species, i.e., white sucker (Catostomus commersoni) and redbreast sunfish (Lepomis auritus), which were the focus of this study. Parasite communities of both hosts were surveyed and compared between each lake and were statistically evaluated for differences in species similarity, prevalence, mean intensity, and mean abundance. Between lakes, white suckers had significant differences in the prevalence of 4 parasite species (Myxobolus sp. 2, Myxobolus bibulatus, Octospinifer macilentis, and Pomphorhynchus bulbocoli) and mean abundances of 4 parasites (neascus larvae, Octospinifer macilentis, Pomphorhynchus bulbocoli, and Glaridacris confusus). Redbreast sunfish had significant differences in the prevalence of 3 species (Myxobolus uvuliferis, a coccidian species, and Spinitectus carolini) and differences in parasite mean abundance of 5 species (neascus larvae, Clinostomum marginatum , Leptorhynchoides thecatus, Spinitectus carolini, and Eustrongylides sp.). Differences in component communities between lakes were found and, although the exact causes cannot be determined by this study, we speculate on several possible explanations.


Evolution of angiosperm seed disperser mutualisms: the timing of origins and their consequences for coevolutionary interactions between angiosperms and frugivores.
The origins of interactions between angiosperms and fruit-eating seed dispersers have attracted much attention following a seminal paper on this topic by Tiffney (1984). This review synthesizes evidence pertaining to key events during the evolution of angiosperm-frugivore interactions and suggests some implications of this evidence for interpretations of angiosperm-frugivore coevolution. The most important conclusions are: (i) the diversification of angiosperm seed size and fleshy fruits commenced around 80 million years ago (Mya). The diversity of seed sizes, fruit sizes and fruit types peaked in the Eocene around 55 to 50 Mya. During this first phase of the interaction, angiosperms and animals evolving frugivory expanded into niche space not previously utilized by these groups, as frugivores and previously not existing fruit traits appeared. From the Eocene until the present, angiosperm-frugivore interactions have occurred within a broad frame of existing niche space, as defined by fruit traits and frugivory, motivating a separation of the angiosperm-frugivore interactions into two phases, before and after the peak in the early Eocene. (ii) The extinct multituberculates were probably the most important frugivores during the early radiation phase of angiosperm seeds and fleshy fruits. Primates and rodents are likely to have been important in the latter part of this first phase. (iii) Flying frugivores, birds and bats, evolved during the second phase, mainly during the Oligocene and Miocene, thus exploiting an existing diversity of fleshy fruits. (iv) A drastic climate shift around the Eocene-Oligocene boundary (around 34 Mya) resulted in more semi-open woodland vegetation, creating patchily occurring food resources for frugivores. This promoted evolution of a 'flying frugivore niche' exploited by birds and bats. In particular, passerines became a dominant frugivore group worldwide. (v) Fleshy fruits evolved at numerous occasions in many angiosperm families, and many of the originations of fleshy fruits occurred well after the peak in the early Eocene. (vi) During periods associated with environmental change altering coevolutionary networks and opening of niche space, reciprocal coevolution may result in strong directional selection formative for both fruit and frugivore evolution. Further evidence is needed to test this hypothesis. Based on the abundance of plant lineages with various forms of fleshy fruits, and the diversity of frugivores, it is suggested that periods of rapid coevolution in angiosperms and frugivores occurred numerous times during the 80 million years of angiosperm-frugivore evolution.


Whole-genome identification of transposable elements reveals the equine repetitive element insertion polymorphism in Chinese horses.
Transposable elements (TEs) are diverse, abundant, and complicated in genomes. They not only can drive the genome evolution process but can also act as special resources for adaptation. However, little is known about the evolutionary processes that shaped horses. In this work, 126 horse assemblages involved in most horse breeds in China were used to investigate the patterns of TE variation for the first time. By using RepeatMasker and melt software, we found that the horse-specific short interspersed repetitive elements family, equine repetitive elements (ERE1), exhibited polymorphisms in horse genomes. Phylogenetic analysis based on these ERE1 loci (minor allele frequency ≥0.05) revealed three major horse groups, namely, those in northern China, southern China, and Qinghai-Tibetan, which mirrors the result determined by SNPs to some extent. The present ERE1 family emerged ~0.26 to 1.77 Mya ago, with an activity peak at ~0.49 Mya, which matches the early stage of the horse lineage and decreases after the divergence of Equus caballus and Equus ferus przewalskii. To detect the functional ERE1(s) associated with adaptation, locus-specific branch length, genome-wide association study, and absolute allele frequency difference analyses were conducted and resulted in two common protein-coding genes annotated by candidate ERE1s. They were clustered into the vascular smooth muscle contraction (p = 0.01, EDNRA) and apelin signalling pathways (p = 0.02, NRF1). Notably, ERE1 insertion into the EDNRA gene showed a higher association with adaptation among southern China horses and other horses in 15 populations and 451 individuals (p = 4.55 e-8). Our results provide a comprehensive understanding of TE variations to analyse the phylogenetic relationships and traits relevant to adaptive evolution in horses.


Functional Characterization of Type III-A CRISPR-Cas in a Clinical Human Methicillin-R Staphylococcus aureus Strain.
CRISPR with its cas genes is an adaptive immune system that protects prokaryotes against foreign genetic elements. The type III-A CRISPR-Cas system is rarely found in Staphylococcus aureus, and little is known about its function in S. aureus. Here, we describe the genome characteristics of the clinical methicillin-resistant S. aureus (MRSA) strain TZ0912, carrying a type III-A CRISPR-Cas system. Phylogenetic analysis of 35 reported CRISPR-Cas-positive S. aureus strains revealed that the CRISPR-Cas system is prevalent in CC8 clones (10/35) and is located in the staphylococcal cassette chromosome mec (SCCmec) V, which confers methicillin resistance. Plasmid transformation and phage infection assays reveal that the type III-A CRISPR-Cas system protects TZ0912 against foreign DNA with sequence homology to the spacers located in the CRISPR array. We observed that the CRISPR-Cas immune system could effectively protect MRSA against phage attacks in both liquid culture and solid medium. In accordance with previous reports, using RNA-seq analysis and plasmid transformation assays, we find that the crRNAs close to the leading sequence of the CRISPR array are more highly expressed and are more effective at directing plasmid elimination compared to the distant spacers. This study established a model for evaluating the efficiency of naive CRISPR-Cas system in MRSA against phage, which could contribute to future research on the function of CRISPR-Cas in clinical MRSA isolates and improve phage therapy against MRSA infections.


Oxidative stress alters mitochondrial bioenergetics and modifies pancreatic cell death independently of cyclophilin D, resulting in an apoptosis-to-necrosis shift.
Mitochondrial dysfunction lies at the core of acute pancreatitis (AP). Diverse AP stimuli induce Ca2+-dependent formation of the mitochondrial permeability transition pore (MPTP), a solute channel modulated by cyclophilin D (CypD), the formation of which causes ATP depletion and necrosis. Oxidative stress reportedly triggers MPTP formation and is elevated in clinical AP, but how reactive oxygen species influence cell death is unclear. Here, we assessed potential MPTP involvement in oxidant-induced effects on pancreatic acinar cell bioenergetics and fate. H2O2 application promoted acinar cell apoptosis at low concentrations (1-10 μm), whereas higher levels (0.5-1 mm) elicited rapid necrosis. H2O2 also decreased the mitochondrial NADH/FAD+ redox ratio and ΔΨm in a concentration-dependent manner (10 μm to 1 mm H2O2), with maximal effects at 500 μm H2O2 H2O2 decreased the basal O2 consumption rate of acinar cells, with no alteration of ATP turnover at <50 μm H2O2 However, higher H2O2 levels (≥50 μm) diminished spare respiratory capacity and ATP turnover, and bioenergetic collapse, ATP depletion, and cell death ensued. Menadione exerted detrimental bioenergetic effects similar to those of H2O2, which were inhibited by the antioxidant N-acetylcysteine. Oxidant-induced bioenergetic changes, loss of ΔΨm, and cell death were not ameliorated by genetic deletion of CypD or by its acute inhibition with cyclosporine A. These results indicate that oxidative stress alters mitochondrial bioenergetics and modifies pancreatic acinar cell death. A shift from apoptosis to necrosis appears to be associated with decreased mitochondrial spare respiratory capacity and ATP production, effects that are independent of CypD-sensitive MPTP formation.


The Therapeutic and Diagnostic Potential of Amyloid β Oligomers Selective Antibodies to Treat Alzheimer's Disease.
Improvements have been made in the diagnosis of Alzheimer's disease (AD), manifesting mostly in the development of in vivo imaging methods that allow for the detection of pathological changes in AD by magnetic resonance imaging (MRI) and positron emission tomography (PET) scans. Many of these imaging methods, however, use agents that probe amyloid fibrils and plaques-species that do not correlate well with disease progression and are not present at the earliest stages of the disease. Amyloid β oligomers (AβOs), rather, are now widely accepted as the Aβ species most germane to AD onset and progression. Here we report evidence further supporting the role of AβOs as pathological instigators of AD and introduce promising anti-AβO diagnostic probes capable of distinguishing the 5xFAD mouse model from wild type mice by PET and MRI. In a developmental study, Aβ oligomers in 5xFAD mice were found to appear at 3 months of age, just prior to the onset of memory dysfunction, and spread as memory worsened. The increase of AβOs is prominent in the subiculum and correlates with concomitant development of reactive astrocytosis. The impact of these AβOs on memory is in harmony with findings that intraventricular injection of synthetic AβOs into wild type mice induced hippocampal dependent memory dysfunction within 24 h. Compelling support for the conclusion that endogenous AβOs cause memory loss was found in experiments showing that intranasal inoculation of AβO-selective antibodies into 5xFAD mice completely restored memory function, measured 30-40 days post-inoculation. These antibodies, which were modified to give MRI and PET imaging probes, were able to distinguish 5xFAD mice from wild type littermates. These results provide strong support for the role of AβOs in instigating memory loss and salient AD neuropathology, and they demonstrate that AβO selective antibodies have potential both for therapeutics and for diagnostics.


Cognitive Fatigue and Processing Speed in Children Treated for Brain Tumours.
The relationship between fatigue and cognition has not been fully elucidated in children and adolescent survivors of brain tumours. The aim of the present study was to investigate the potential relationship between fatigue and cognitive impairments in these survivors, as this group is at risk for both types of deficits. Survivors of paediatric brain tumours (n = 45) underwent a neuropsychological testing on average 4 years after diagnosis. Mean age at follow-up was 13.41 years. Cognition was assessed with neuropsychological tests, and fatigue with the Pediatric Quality of Life (PedsQL™) Multidimensional Fatigue Scale. Regression analysis, adjusted for cranial radiotherapy and age at diagnosis, was used to investigate the associations between cognitive variables and fatigue subscales. Cognitive variables associated with fatigue were subsequently exploratively assessed. Significant associations were found for cognitive fatigue and measures of cognitive processing speed; Coding: p = .003, r = .583, 95% CI [9.61; 22.83] and Symbol Search: p = .001, r = .585, 95% CI [10.54; 24.87]. Slower processing speed was associated with poorer results for cognitive fatigue. Survivors with the largest decrease in processing speed from baseline to follow-up also experienced the most cognitive fatigue. Survivors expressed more cognitive fatigue compared to other types of fatigue. The association between cognitive fatigue and cognitive processing speed in children and adolescents treated for brain tumours is in concordance with the results previously reported in adults. Some survivors experience fatigue without impairment in processing speed, indicating the need for comprehensive assessments. Moreover, the study supports that fatigue is a multidimensional concept which should be measured accordingly.


IRE1-bZIP60 Pathway Is Required for Nicotiana attenuata Resistance to Fungal Pathogen Alternaria alternata.
As an endoplasmic reticulum (ER) stress sensor, inositol-requiring enzyme 1 (IRE1) splices the bZIP60 mRNA, and produces an active bZIP60 transcription factor that regulates genes involved in the unfolded protein response (UPR) during ER stresses. This IRE1-bZIP60 pathway is conserved in plant species and recently implicated in plant-pathogen interaction. However, it is unclear whether this IRE1-bZIP60 pathway is involved in Nicotiana attenuata resistance to necrotic fungal pathogen, Alternaria alternata. In this study, transcriptional levels of chaperone protein genes, including luminal binding protein (BiP), protein disulfide isomerase (PDI), calnexin 1-like (CNX 1-like), and calreticulin (CRT), and genes involved in IRE1-bZIP60 pathway, were all significantly induced in N. attenuata leaves after A. alternata inoculation. Silencing IRE1 or bZIP60 led to N. attenuata plants more susceptible to A. alternata, which were associated with reduced gene expressions of Feruloyl-CoA 6'-hydroxylase 1 (F6'H1), a gene encoding a key enzyme for phytoalexin scopoletin and scopolin biosynthesis. Further, electromobility shift assays (EMSA) indicated that bZIP60 protein of spliced form could directly bind to the promoter region of F6'H1 in vitro. JA signaling pathway is required for N. attenuata resistance to A. alternata. Interestingly, the fungus-elicited transcriptional levels of BiP, PDI, CNX 1-like, CRT, IRE1, and bZIP60(s) were all significantly decreased in JA-deficient or JA-insensitive plants. Meanwhile, those genes were significantly induced by methyl jasmonate (MeJA) when applied exogenously. However, the transcriptional levels of JA-regulated genes allene oxide synthase (AOS) and lipoxygenease 3 (LOX3) were not affected in plants impaired with IRE1-bZIP60 pathway. Thus, it is concluded that IRE1-bZIP60 pathway is required for N. attenuata resistance to A. alternata, and JA signaling pathway plays an important role in the elicitation of chaperone protein genes and IRE1-bZIP60 pathway.


Combinatorial co-encapsulation of hydrophobic molecules in poly(lactide-co-glycolide) microparticles.
There is great interest for developing poly(lactide-co-glycolide) (PLGA) based particles for targeted delivery and controlled release of encapsulated biological molecules. These PLGA particles can be used to deliver proteins, small molecule drugs and nucleotides. Furthermore, it has been shown that the co-encapsulation of multiple factors in PLGA particles can generate synergistic responses, and can also provide theranostic capability. However, the number of possible unique particle formulations that may be generated by the combination of different components in a particle increases dramatically with each new component, and currently, there is no method to generate large libraries of unique PLGA particles. In order to address this gap, we have developed a high-throughput methodology to produce hundreds of small batches of particles. The particles are generated in multi-well plate wells by a modified oil-in-water emulsion technique. In order to demonstrate the versatility of this technique, combinatorial formulations of six different loading concentrations of three fluorescent dyes were fabricated giving rise to 216 unique PLGA particle formulations. We demonstrate systematic and well-controlled combinatorial loading of hydrophobic molecules into the particles. This parallel particle production (PPP) methodology potentiates the generation of hundreds of different combinatorial particle formulations with multiple co-encapsulates in less than 24 h in standard polystyrene multi-well plates, thus providing rapid, low cost, high-throughput production. We envision that such a PPP library of particles encapsulating combinations of drugs and imaging modalities can subsequently be tested on small populations of cells in a high-throughput fashion, and represents a step toward personalized medicine.


Breath Figures of Nanoscale Bricks: A Universal Method for Creating Hierarchic Porous Materials from Inorganic Nanoparticles Stabilized with Mussel-Inspired Copolymers.
High-performance catalysts and photovoltaics are required for building an environmentally sustainable society. Because catalytic and photovoltaic reactions occur at the interfaces between reactants and surfaces, the chemical, physical, and structural properties of interfaces have been the focus of much research. To improve the performance of these materials further, inorganic porous materials with hierarchic porous architectures have been fabricated. The breath figure technique allows preparing porous films by using water droplets as templates. In this study, a valuable preparation method for hierarchic porous inorganic materials is shown. Hierarchic porous materials are prepared from surface-coated inorganic nanoparticles with amphiphilic copolymers having catechol moieties followed by sintering. Micron-scale pores are prepared by using water droplets as templates, and nanoscale pores are formed between the nanoparticles. The fabrication method allows the preparation of hierarchic porous films from inorganic nanoparticles of various shapes and materials.


Mode of inhibition of HIV reverse transcriptase by 2-hexaprenylhydroquinone, a novel general inhibitor of RNA-and DNA-directed DNA polymerases.
A natural compound from the Red Sea sponge Ircinia sp., 2-hexaprenylhydroquinone (HPH), has been shown to be a general inhibitor of retroviral reverse transcriptases (from HIV-1, HIV-2 and murine leukaemia virus) as well as of cellular DNA polymerases (Escherichia coli DNA polymerase I, and DNA polymerases alpha and beta). The pattern of inhibition was found to be similar for all DNA polymerases tested. Thus the mode of inhibition was studied in detail for HIV-1 reverse transcriptase. HPH is a non-competitive inhibitor and binds the enzyme irreversibly with high affinity (Ki=0. 62 microM). The polar hydroxy groups have been shown to be of key importance. A methylated derivative, mHPH, which is devoid of these polar moieties, showed a significantly decreased capacity to inhibit all DNA polymerases tested. Like the natural product, mHPH binds the enzyme independently at an allosteric site, but with reduced affinity (Ki=7.4 microM). We show that HPH does not interfere with the first step of the polymerization process, i.e. the physical formation of the reverse-transcriptase-DNA complex. Consequently, we suggest that the natural inhibitor interferes with the subsequent steps of the overall reaction. Since HPH seems not to affect the affinity of dNTP for the enzyme (the Km is unchanged under conditions where the HPH concentration is increased), we speculate that its inhibitory capacity is derived from its effect on the nucleotidyl-transfer catalytic reaction. We suggest that such a mechanism of inhibition is typical of an inhibitor whose mode of inhibition should be common to all RNA- and DNA-directed polymerases.


Assessing the Effects of Land Use on Surface Water Quality in the Lower uMfolozi Floodplain System, South Africa.
This study investigated the impacts of cultivation on water and soil quality in the lower uMfolozi floodplain system in KwaZulu-Natal province, South Africa. We did this by assessing seasonal variations in purposefully selected water and soil properties in these two land-use systems. The observed values were statistically analysed by performing Student's paired t-tests to determine seasonal trends in these variables. Results revealed significant seasonal differences in chloride and sodium concentrations and electrical conductivity (EC) and the sodium adsorption ratio (SAR) with cultivated sites exhibiting higher values. Most of the analyzed chemical parameters were within acceptable limits specified by the South African agricultural-water-quality (SAWQ) water quality guidelines for irrigation except for sodium adsorption ratio (SAR), chloride, sodium and EC. EC, pH and nitrate content which were higher than the specified SAWQ limits in cultivated sites. Quantities of glyphosate, ametryn and imidacloprid could not be measured because they were below detectable limits. The study concludes that most water quality parameters met SAWQ's standards. These results argue for concerted efforts to systematically monitor water and soil quality characteristics in this environment to enhance sustainability by providing timely information for management purposes.


Hyper-IgM syndrome--a case report and a clinical perspective.
The authors describe the case of a 28-year-old woman, with a history of recurrent bacterial infections since childhood and multiple hospitalizations for pneumonia, with important pulmonary sequelae, including bronchiectasis which warranted the need to perform a left lobectomy and lingulectomia at age 13. After diagnostic work up, the diagnosis of hypogammaglobulinemia with hyper-IgM was established, and she began regular replacement i.v. immunoglobulin treatment, with good tolerance and no side effects. A sequencing of the entire coding region (exons 1-5) of the AICDA gene was performed, and a homozygous c.260G > C mutation was identified, confirming the diagnosis of type 2 hyper-IgM syndrome. This case illustrates the negative impact that a delay in diagnosis and hence delay in treatment has in patients with primary immunodeficiency since early therapy is the only way to reduce the incidence and severity of complications.


Published randomized controlled trials of surveillance in cancer patients - a systematic review.
With solid tumor cancer survivorship increasing, the number of patients requiring post-treatment surveillance also continues to increase. This highlights the need for evidence-based cancer surveillance guidelines. Ideally, these guidelines would be based on combined high-quality data from randomized controlled trials (RCTs). We present a systematic review of published cancer surveillance RCTs in which we sought to determine the feasibility of data pooling for guideline development. We carried out a systematic search of medical databases for RCTs in which adult patients with solid tumors that had undergone surgical resection with curative intent and had no metastatic disease at presentation, were randomized to different surveillance regimens that assessed effectiveness on overall survival (OS). We extracted study characteristics and primary and secondary outcomes, and assessed risk of bias and validity of evidence with standardized checklist tools. Our search yielded 32,216 articles for review and 18 distinct RCTs were included in the systematic review. The 18 trials resulted in 23 comparisons of surveillance regimens. There was a highlevel of variation between RCTs, including the study populations evaluated, interventions assessed and follow-up periods for the primary outcome. Most studies evaluated colorectal cancer patients (11/18, [61%]). The risk of bias and validity of evidence were variable and inconsistent across studies. This review demonstrated that there is tremendous heterogeneity among RCTs that evaluate effectiveness of different postoperative surveillance regimens in cancer patients, rendering the consolidation of data to inform high-quality cancer surveillance guidelines unfeasible. Future RCTs in the field should focus on consistent methodology and primary outcome definition.


[Expression of the cerbB-2 (HER-2/neu) oncoprotein in prostatic adenocarcinoma].
Our aim is to determine the expression of the cerbB-2 oncoprotein in prostate cancers using an immunohistochemistry staining and to compare these results with several clinical and histological prognostic factors. An immunohistochemical staining using the cerbB-2 monoclonal antibody (Dako) was performed in 32 radical prostatectomy specimens diagnosed of adenocarcinoma. The intensity of cerbB-2 expression was evaluated with a scale that variated from 0 (no staining) to 3+ (strong complete membrane staining) according to published guidelines. Association of cerbB-2 index immunoreactivity with clinical and histological prognostic factors was examined. Definite positive membranous staining was detected in 14 of 32 neoplastic cases (44%). Such overexpression was correlated with higher Gleason grade (p=0.04) and higher stage of disease (p=0.038). 1) This study shows that 44% of all prostate cancer express the cerbB-2 oncoprotein with immunohistochemical technique. 2) These findings suggest that is necessary to standardize the immunohistochemical staining procedure with cerbB-2 in prostate adenocarcinoma. 3) The level of cerbB-2 expression was correlated with Gleason grade and clinical stage.


An Online International Collaborative Learning Program During the COVID-19 Pandemic for Nursing Students: Mixed Methods Study.
Given the limitations imposed by the COVID-19 pandemic, a better understanding of how nursing programs around the globe have implemented distance education methods and related initiatives to provide international collaborative learning opportunities as well as complementary aspects of practical education would be constructive for nursing students. It is expected that international collaboratives through web-based communication will continue to be increasingly utilized after the pandemic; therefore, it is time to discuss the effects and direction of these developments. We aimed to examine the impact of an online international collaborative learning program on prelicensure nursing students' international and global competencies in South Korea. We conducted a mixed methods study (web-based surveys and focus group interviews). A total of 15 students participated in the study. The surveys were used to examine changes in participants' global leadership competencies, and the focus group interviews were used to evaluate the program's effectiveness and to identify opportunities for improvement. The online international collaborative program consisted of 7 synchronous web-based classroom sessions. Each session ran for 60 to 90 minutes. Faculty experts and nurses working in the United States discussed various topics with students, such as nursing education in the United States and evidence-based teaching and learning. The students gave presentations on the South Korean nursing education system. Data were analyzed with descriptive statistics, the Mann-Whitney U test, and content analysis methods. Participants reported improvement in their global leadership competencies. Four main categories emerged from analysis of the focus interviews: (1) realistic applicability, (2) clarification, (3) expansion of perspectives, and (4) initiative. The online international collaborative learning program had a positive impact on the development of students' international competencies. The findings support the further development of international exchange programs through web-based meetings in the postpandemic era.


Highly sensitive serological approaches for Pepino mosaic virus detection.
Pepino mosaic virus (PepMV) causes severe disease in tomato and other Solanaceous crops around globe. To effectively study and manage this viral disease, researchers need new, sensitive, and high-throughput approaches for viral detection. In this study, we purified PepMV particles from the infected Nicotiana benthamiana plants and used virions to immunize BALB/c mice to prepare hybridomas secreting anti-PepMV monoclonal antibodies (mAbs). A panel of highly specific and sensitive murine mAbs (15B2, 8H6, 23D11, 20D9, 3A6, and 8E3) could be produced through cell fusion, antibody selection, and cell cloning. Using the mAbs as the detection antibodies, we established double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), Dot-ELISA, and Tissue print-ELISA for detecting PepMV infection in tomato plants. Resulting data on sensitivity analysis assays showed that both DAS-ELISA and Dot-ELISA can efficiently monitor the virus in PepMV-infected tissue crude extracts when diluted at 1:1 310 720 and 1:20 480 (weight/volume ratio (w/v), g/mL), respectively. Among the three methods developed, the Tissue print-ELISA was found to be the most practical detection technique. Survey results from field samples by the established serological approaches were verified by reverse transcription polymerase chain reaction (RT-PCR) and DNA sequencing, demonstrating all three serological methods are reliable and effective for monitoring PepMV. Anti-PepMV mAbs and the newly developed DAS-ELISA, Dot-ELISA, and Tissue print-ELISA can benefit PepMV detection and field epidemiological study, and management of this viral disease, which is already widespread in tomato plants in Yunnan Province of China.


Fluorometric evaluation of large and small intestinal ischaemia in the horse.
Arteriovenous (ischaemic strangulating obstruction, ISO) and venous obstructions (haemorrhagic strangulating obstruction, HSO) were created for 70 min in the small intestine of eight anaesthetised horses, and ISO was created in four horses for 2 h and four horses for 3 h at the sternal and diaphragmatic flexures of the large colon. Five minutes following release of the occlusions, sodium fluorescein 20 per cent (0.5 mg/kg bodyweight intravenously) was administered. Serial quantitative measurements of serosal surface fluorescence of the injured segments and a control segment were made at 2, 10, 15, 20 and 30 mins using a fibreoptic perfusion fluorometer. Significant differences in fluorescence were seen in small intestine segments between HSO vs control, ISO vs HSO, but not ISO vs control segments. In the large colon, significant differences were seen between the fluorescence in the ischaemic 3 h ventral colon and control segments, and a trend for difference in fluorescence between the 2 and 3 h ischaemic segments. Differences in fluorescence between 2 vs 3 h dorsal colon segments and 2 or 3 h dorsal colon vs control segments were not significant.


Trefoil factor 2 negatively regulates type 1 immunity against Toxoplasma gondii.
IL-12-mediated type 1 inflammation confers host protection against the parasitic protozoan Toxoplasma gondii. However, production of IFN-γ, another type 1 inflammatory cytokine, also drives lethality from excessive injury to the intestinal epithelium. As mechanisms that restore epithelial barrier function following infection remain poorly understood, this study investigated the role of trefoil factor 2 (TFF2), a well-established regulator of mucosal tissue repair. Paradoxically, TFF2 antagonized IL-12 release from dendritic cells (DCs) and macrophages, which protected TFF2-deficient (TFF2(-/-)) mice from T. gondii pathogenesis. Dysregulated intestinal homeostasis in naive TFF2(-/-) mice correlated with increased IL-12/23p40 levels and enhanced T cell recruitment at baseline. Infected TFF2(-/-) mice displayed low rates of parasite replication and reduced gut immunopathology, whereas wild-type (WT) mice experienced disseminated infection and lethal ileitis. p38 MAPK activation and IL-12p70 production was more robust from TFF2(-/-)CD8+ DC compared with WT CD8+ DC and treatment of WT DC with rTFF2 suppressed TLR-induced IL-12/23p40 production. Neutralization of IFN-γ and IL-12 in TFF2(-/-) animals abrogated resistance shown by enhanced parasite replication and infection-induced morbidity. Hence, TFF2 regulated intestinal barrier function and type 1 cytokine release from myeloid phagocytes, which dictated the outcome of oral T. gondii infection in mice.


Costs and renumeration of osteomyelitis treatment involving free flaps: implications of return to theatre.
Aim: This study aimed to define the costs of surgical management of chronic osteomyelitis where free tissue transfer was required in addition to debridement of bone, particularly the increased costs incurred by a return to theatre. We hypothesised that there would be a significantly greater cost when patients required re-exploration for vascular compromise. Method: We retrospectively analysed the costs of a consecutive series of sixty patient episodes treated at the Bone Infection Unit in Oxford from 2012 to 2015. Treatment involved excision of osteomyelitis with free tissue transfer for immediate soft tissue cover. We compared the costs of uncomplicated cases with those who returned to theatre and determined the profit / loss for the hospital from renumeration through the UK National Health Service Tariff Structure. Results: Hospital income according to UK HRG tariff was compared to the actual cost of treatment and these 60 cases were significantly underfunded overall (P < 0.005). In just 1 case, the cost to the hospital was completely covered by tariff. Six patients (10%) returned to theatre for urgent flap re-exploration with five flaps salvaged and one failed, requiring another free flap reconstruction (1.7%). These six patient episodes had a significantly higher mean cost compared to the uncomplicated cases. The average financial loss to the hospital for patients who did return to theatre was £19401 (range £8103 to £48380) and in those who did not was £9600 (range - £600 to £23717). The case requiring further free tissue transfer cost a total of £74158, £48380 more than the hospital was paid: the most extreme discrepancy. The overall loss for this group of 60 patients was £610 090. Conclusions: Surgery for chronic osteomyelitis is multidisciplinary, complex and therefore expensive with a significant risk of complications. However, this study demonstrates that the hospital currently makes a financial loss on almost all patients but especially if flap complications occur. This study has implications for the long term viability of specialist units treating this important disease.


Study on the Effect of Bushen Zhuanggu Tablet Combined with Conventional Regimen on Bone Mineral Density Improvement, Functional Recovery and Fracture Risk Prevention in Patients with Postmenopausal Osteoporosis.
This case-control study was to explore the effect of Bushen Zhuanggu tablet combined with routine regimen on bone mineral density (BMD) improvement, functional recovery, and fracture prevention in postmenopausal osteoporosis (PMOP) patients. 180 postmenopausal osteoporosis patients were randomly selected from communities A, B, and C cohorts as research subjects from January to May 2021. The study subjects were divided into three groups. The groups were in a 1 : 1 ratio according to the principles of nonrandomised, concurrent controlled trials, and methods. There were 60 participants in each group (group A, group B, and group C). Group A was treated with Bushen Zhuanggu tablet for antiosteoporosis + basic treatment (calcium supplement and vitamin D). Group C was given Bushen Zhuanggu tablet for antiosteoporosis intervention. Group B was given basic treatment (calcium supplement and vitamin D supplementation) as a control group. The follow-up time was 6 months after treatment. Finally, we compare the differences in calcium and phosphorus metabolism indexes, BMD, bone metabolism indexes, upper and lower limb muscle strength, and quality of life scores. Group A, B, and C's effective rate was 98.33%, 80.00%, and 93.33%, respectively. The group A's effective rate was significantly higher than that in group B and C, and the difference was statistically significant (P < 0.05). After 6 months intervention, the levels of serum Ca2+, serum phosphorus (P), serum creatinine (Cr), and parathyroid hormone (PTH) in 3 groups decreased. Ca, P, Scr, and PTH levels in group A were the lowest among study groups, and the difference was statistically significant (P < 0.05). The increase in the BMD of lumbar spine, the left femoral neck, and Ward's triangle area of the three groups were observed with the highest data in group A. After 6 months of treatment, the levels of serum N-terminal propeptide of type I procollagen, PINP, and serum osteocalcin (OC) increased, while the levels of β-cross-linked C-terminal telopeptide of type I collagen (β-CTX) and alkaline phosphatase (ALP) decreased in the three groups. The improvement of all bone metabolic indexes in group A was significantly better than that in B and C groups, and the difference was statistically significant (P < 0.05). The enhanced upper limb muscle strength and the shorter standing-walking timing test (TUGT) time were observed after 6 months of treatment. The improvement effect of upper and lower limb muscle strength in group A was significantly better than that in B and C groups, and the difference was statistically significant (P < 0.05). There were significant differences in physiological function, life function, general health status, physical pain, mental state, emotional function, vitality, and social function among the three groups after 6 months treatment, and the difference was statistically significant (P < 0.05). The score of quality of life in group A was higher than that in B and C groups, and the difference was statistically significant (P < 0.05). Bushen Zhuanggu tablet combined with conventional therapy is effective in the postmenopausal osteoporosis treatment, which effectively increase the BMD, regulate calcium and phosphorus metabolism, promote the recovery of limb function, prevent the recurrence of fracture, and improve the patients' quality of life. This treatment scheme is worth popularizing.


Mosquito NADPH-cytochrome P450 oxidoreductase: kinetics and role of phenylalanine amino acid substitutions at leu86 and leu219 in CYP6AA3-mediated deltamethrin metabolism.
The NADPH-cytochrome P450 oxidoreductase (CYPOR) enzyme is a membrane-bound protein and contains both FAD and FMN cofactors. The enzyme transfers two electrons, one at a time, from NADPH to cytochrome P450 enzymes to function in the enzymatic reactions. We previously expressed in Escherichia coli the membrane-bound CYPOR (flAnCYPOR) from Anopheles minimus mosquito. We demonstrated the ability of flAnCYPOR to support the An. minimus CYP6AA3 enzyme activity in deltamethrin degradation in vitro. The present study revealed that the flAnCYPOR purified enzyme, analyzed by a fluorometric method, readily lost its flavin cofactors. When supplemented with exogenous flavin cofactors, the activity of flAnCYPOR-mediated cytochrome c reduction was increased. Mutant enzymes containing phenylalanine substitutions at leucine residues 86 and 219 were constructed and found to increase retention of FMN cofactor in the flAnCYPOR enzymes. Kinetic study by measuring cytochrome c-reducing activity indicated that the wild-type and mutant flAnCYPORs followed a non-classical two-site Ping-Pong mechanism, similar to rat CYPOR. The single mutant (L86F or L219F) and double mutant (L86F/L219F) flAnCYPOR enzymes, upon reconstitution with the An. minimus cytochrome P450 CYP6AA3 and a NADPH-regenerating system, increased CYP6AA3-mediated deltamethrin degradation compared to the wild-type flAnCYPOR enzyme. The increased enzyme activity could illustrate a more efficient electron transfer of AnCYPOR to CYP6AA3 cytochrome P450 enzyme. Addition of extra flavin cofactors could increase CYP6AA3-mediated activity supported by wild-type and mutant flAnCYPOR enzymes. Thus, both leucine to phenylalanine substitutions are essential for flAnCYPOR enzyme in supporting CYP6AA3-mediated metabolism.


SOX5 interacts with YAP1 to drive malignant potential of non-small cell lung cancer cells.
The dysregulation of transcription factors plays a vital role in tumor initiation and progression. Sex determining region Y-box 5 (SOX5) encodes a member of the SRY-related HMG-box family of transcription factors involved in the determination of the cell fate and the regulation of embryonic development. However, its functional roles in non-small cell lung cancer (NSCLC) remain unclear. Herein, we report that SOX5 sustains stem-like traits and enhances the malignant phenotype of NSCLC cells. We determine that SOX5 is preferentially expressed by cancer stem-like cells (CSLCs) of human NSCLC. In vitro gain- and loss-of-function studies demonstrate that SOX5 promotes self-renewal, invasion and migration in NSCLC cells. Importantly, knockdown of SOX5 potently inhibits tumor growth in a xenograft mouse model. Mechanistically, YAP1 can act as an interacting protein of SOX5 to drive the malignant potential of NSCLC cells. Silencing of YAP1 attenuates the malignant processes in NSCLC cells, which is consistent with the function of SOX5 loss. SOX5 overexpression reverses the attenuated malignant progression in YAP1 knockdown cancer cells. Taken together, these findings identify that SOX5 acts as an oncogenic factor by interacting with YAP1 in NSCLC cells and may be a potential therapeutic target for NSCLC patients.


Patterns of cervical spine injury and their associated lesions.
Motorcycle riding and diving into shallow water continue to present a high risk of cervical spine injury, often complicated by spinal cord damage. In patients with high cervical cord trauma, differentiation of arterial hypotension due to losing vasomotor control from the effects of internal hemorrhage can cause difficulty. In a series of 123 consecutive cases of cervical spine injury, no evidence was found that either early surgical treatment or steroid administration exert a favorable influence on recovery from traumatic myelopathy. When compared with other series, differences were found in the nature, frequency and severity of both spinal and associated injuries, resulting from the relative frequency among the population studied of trauma due to a particular mechanism-traffic accident, diving, industrial injury-and the special functions and location of the hospital from which information is gathered.


Impact of Top Electrodes on the Nonvolatile Resistive Switching Properties of Citrus Thin Films.
Natural citrus thin films on an indium tin oxide (ITO)/glass substrate were synthesized using the solution method for resistive random access memory (RRAM) applications. The results indicated that the citrus memory device possessed stable resistive switching behavior. For a clear understanding of the role of the interface reaction between the top metal electrode and the citrus film, we investigated the influences of various top electrode (TE) materials on the resistive switching in TE/citrus/ITO devices. In comparison with Au/citrus/ITO and Ti/citrus/ITO devices, the Al/citrus/ITO device can be reproduced with a DC voltage of more than 100 times while only showing a slight decrease in the ON/OFF ratio. In addition, the Al/citrus/ITO device exhibited a high ON/OFF ratio of over 104 and an outstanding uniformity, which was attributed to the fast formation of a native oxide layer (AlOx), as confirmed by the line scan analysis. This indicated that the interface layer, created by the redox reaction between the Al electrode and citrus film, played an important role in the resistive switching properties of TE/citrus/ITO structures. These findings can serve as design guidelines for future bio-based RRAM devices.


Epidemiology of renal dysfunction and patient outcome in atherosclerotic renal artery occlusion.
Patients with atherosclerotic renal artery occlusion (RAO) effectively have only a single functioning kidney, so they constitute an ideal group in whom to study the relationship of atherosclerotic renovascular disease (ARVD) severity to renal functional outcome. Of 299 patients with ARVD who had presented to a single center over a 12-yr period, 142 (47.5%) patients with RAO were identified. There was no relationship between baseline renal function and contralateral renovascular anatomy. Patients with contralateral normal, insignificant (<50%), or significant (>50%) renal artery stenoses had baseline creatinine of 243 +/- 235, 292 +/- 197, or 210 +/- 102 micromol/L, respectively, but patients with bilateral RAO (creatinine, 540 +/- 304 micromol/L; P < 0.0001) were significantly worse. There were significant correlations between baseline GFR and both proteinuria (r = -0.32; P < 0.01) and contralateral bipolar renal length (r = 0.44; P < 0.0001). Over a mean follow-up period of 31 +/- 21 (2 to 82) mo, the overall rate of progressive renal functional decline was -4.1 ml/min per yr. Nine patients required dialysis at presentation and a further 15 (10.5%) during the course of the study. There were 85 (59.9%) deaths; median survival of the whole group was 25 mo, and 5-yr survival was 31%. Multivariate analysis indicated that low baseline GFR was the chief variable independently associated with increased probability of death or need of dialysis but that renal vascular anatomy had no prognostic impact. This study reinforces the importance of intrarenal vascular and parenchymal disease in the etiology of renal dysfunction in ARVD.


Dry eye modifies the thermal and menthol responses in rat corneal primary afferent cool cells.
Dry eye syndrome is a painful condition caused by inadequate or altered tear film on the ocular surface. Primary afferent cool cells innervating the cornea regulate the ocular fluid status by increasing reflex tearing in response to evaporative cooling and hyperosmicity. It has been proposed that activation of corneal cool cells via a transient receptor potential melastatin 8 (TRPM8) channel agonist may represent a potential therapeutic intervention to treat dry eye. This study examined the effect of dry eye on the response properties of corneal cool cells and the ability of the TRPM8 agonist menthol to modify these properties. A unilateral dry eye condition was created in rats by removing the left lacrimal gland. Lacrimal gland removal reduced tears in the dry eye to 35% compared with the contralateral eye and increased the number of spontaneous blinks in the dry eye by over 300%. Extracellular single-unit recordings were performed 8-10 wk following surgery in the trigeminal ganglion of dry eye animals and age-matched controls. Responses of corneal cool cells to cooling were examined after the application of menthol (10 μM-1.0 mM) to the ocular surface. The peak frequency of discharge to cooling was higher and the cooling threshold was warmer in dry eye animals compared with controls. The dry condition also altered the neuronal sensitivity to menthol, causing desensitization to cold-evoked responses at concentrations that produced facilitation in control animals. The menthol-induced desensitization of corneal cool cells would likely result in reduced tearing, a deleterious effect in individuals with dry eye.


Countrywide inter-epidemic region migration pattern suggests the role of southwestern population in wheat stripe rust epidemics in China.
Understanding countrywide pathogen population structure and inter-epidemic region spread is crucial for deciphering crop potential losses. Wheat stripe rust caused by Puccinia striiformis f. sp. tritici is a destructive disease that affects worldwide wheat production, widespread in China, representing largest epidemic region globally. This study aimed to understand the population structure and migration route of P. striiformis f. sp. tritici across China based on sampling from 15 provinces representing six epidemic zones, viz., over-summering, over-wintering, eastern, Yun-Gui, Xinjiang and Tibet epidemic regions. High genotypic diversity was recorded in over-summering, Tibet and over-wintering epidemic regions. Epidemic regions partly explain population subdivision with variable divergence (FST = 0.005-0.344). Xinjiang and Tibet epidemic regions were independent epidemic zones with least sharing of genotypes. Among other epidemic zones, i.e. over-summering, over-wintering, eastern and Yun-Gui epidemic zones, re-sampling MLGs, clustering-based structure, DAPC analyses, relative migration and low divergence (FST from 0.006 to 0.073) revealed frequent geneflow. Yun-Gui epidemic regions, with a potential for both over-summering and over-wintering, could play an important role in causing epidemics in main wheat-cultivating areas of China. High diversity, recombination signatures and inter-epidemic region migration patterns need to be considered in host-resistant cultivar development in China and neighbouring countries, considering risk of long-distance migration capacity of pathogen.


Exfoliation Syndrome: A Disease of Autophagy and LOXL1 Proteopathy.
Exfoliation syndrome (XFS) is an age-related disease involving the deposition of aggregated fibrillar material (exfoliation material) at extracellular matrices in tissues that synthesize elastic fibers. Its main morbidity is in the eye, where exfoliation material accumulations form on the surface of the ciliary body, iris, and lens. Exfoliation glaucoma (XFG) occurs in a high proportion of persons with XFS and can be a rapidly progressing disease. Worldwide, XFG accounts for about 25% of open-angle glaucoma cases. XFS and XFG show a sharp age-dependence, similarly to the many age-related diseases classified as aggregopathies. Progress in understanding the cellular bases for XFS/XFG has been slowed by a lack of experimental models. Working with primary human tenon fibroblasts (TF) derived from trabeculectomies of XFG patients and age-matched primary open-glaucoma controls, we found that TF from XFG cells display many of the functional features observed in cells from other protein aggregate diseases, such as Parkinson, Alzheimer, Huntington, and age-related macular degeneration. We have documented defects in lysosomal positioning, microtubule organization, autophagy processing rate, and mitochondrial health. In regard to failure of lysosomal and autophagosome positioning in XFG cells, we have found that XFG TF are unable to establish the transnuclear microtubule organizing center that is required for efficient centripetal vesicular locomotion along microtubules. In regard to potential sources of the autophagy malfunction, we have directed our attention to a potential role of the lysyl oxidase-like 1 protein (LOXL1), the elastic fiber catalyst that displays variant-dependent association with risk for XFG. Our experiments show that (a) in XFG cells, a substantial fraction of LOXL1 is processed for degradation by the autophagic system; (b) most of the LOXL1 N-terminus domain exists in a highly disordered state, a condition known to greatly increase the frequency of polypeptide misfolding; (c) that maximum misfolding occurs at amino acid position 153, the location of the high risk variant G153D; and (d) that replacement of glycine (G) by aspartate (D) there results in a substantial decrease in disorder within the 20 amino acid surrounding domain. Finally, we show that clusterin, a protein that can be induced by the presence of intracellular, or extracellular aggregates, is uniformly overexpressed in XFG TF. The implications of our results for a theory relating XFG to cellular aggregopathy are discussed.


Is activation of transversus abdominis and obliquus internus abdominis associated with long-term changes in chronic low back pain? A prospective study with 1-year follow-up.
To investigate associations between deep abdominal muscle activation and long-term pain outcome in chronic non-specific low back pain (LBP). Recruitment of transversus abdominis and obliquus internus abdominis during the abdominal drawing-in manoeuvre was recorded by B-mode ultrasound and anticipatory onset of deep abdominal muscle activity with M-mode ultrasound. Recordings were done before and after 8 weeks with guided exercises for 109 patients with chronic non-specific LBP. Pain was assessed with a numeric rating scale (0-10) before and 1 year after intervention. Associations between muscle activation and long-term pain were examined by multiple linear and logistic regression methods. Participants with a combination of low baseline lateral slide in transversus abdominis and increased slide after intervention had better odds for long-term clinically important pain reduction (≥2 points on the numeric rating scale) compared with participants with small baseline slide and no improvement in slide (OR 14.70, 95% CI 2.41 to 89.56). There were no associations between contraction thickness ratios in transversus abdominis or obliquus internus abdominis and pain at 1-year follow-up. Transversus abdominis lateral slide before intervention was marginally associated with a lower OR for clinically important improvement in pain at 1-year follow-up (OR 0.76, 95% CI 0.62 to 0.93). Delayed onset of the abdominal muscles after the intervention period was weakly associated with higher long-term pain. Improved transversus abdominis lateral slide among participants with low baseline slide was associated with clinically important long-term pain reduction. High baseline slide and delayed onset of abdominal muscles after the intervention period were weakly associated with higher pain at 1-year follow-up. Clinical Trial Registration number The study was preregistered in ClinicalTrials.gov with identifier NCT00201513.


Insomnia and circadian sleep disorders in ovarian cancer: Evaluation and management of underestimated modifiable factors potentially contributing to morbidity.
Ovarian cancer is the leading cause of gynaecological cancer deaths and the seventh most commonly diagnosed cancer among women worldwide, so that, as it is related to substantial and increasing disease burden, the management of ovarian cancer survivors should be a priority. Such issues involve prevention and management of emotional distress, anxiety/depressive symptoms, and maintenance of quality of life from initial diagnosis to post-treatment. Within this framework, sleep disturbances, in particular insomnia, are emerging as modifiable determinants of mental health, also contributing to substantial morbidity among cancer, including ovarian cancer. To this aim we conducted a systematic review according to PRISMA guidelines on prevalence and management of insomnia and circadian sleep disorders in ovarian cancer, while selecting 22 papers. Insomnia was evaluated in ovarian cancer and, while circadian sleep disturbances were poorly assessed in ovarian cancer, insomnia increased from 14% to 60% of patients. Insomnia was associated with cancer-related comorbid conditions such as emotional distress, anxiety/depressive symptoms and low quality of life. Despite this evidence, no studies have been conducted about insomnia treatment in ovarian cancer. The burden of insomnia and circadian sleep disorders in patients with ovarian cancer still needs to be addressed, and requires a call to action for the evaluation and management of these potential modifiable factors that might contribute to ovarian cancer morbidity.


ChimeraTE: a pipeline to detect chimeric transcripts derived from genes and transposable elements.
Transposable elements (TEs) produce structural variants and are considered an important source of genetic diversity. Notably, TE-gene fusion transcripts, i.e. chimeric transcripts, have been associated with adaptation in several species. However, the identification of these chimeras remains hindered due to the lack of detection tools at a transcriptome-wide scale, and to the reliance on a reference genome, even though different individuals/cells/strains have different TE insertions. Therefore, we developed ChimeraTE, a pipeline that uses paired-end RNA-seq reads to identify chimeric transcripts through two different modes. Mode 1 is the reference-guided approach that employs canonical genome alignment, and Mode 2 identifies chimeras derived from fixed or insertionally polymorphic TEs without any reference genome. We have validated both modes using RNA-seq data from four Drosophila melanogaster wild-type strains. We found ∼1.12% of all genes generating chimeric transcripts, most of them from TE-exonized sequences. Approximately ∼23% of all detected chimeras were absent from the reference genome, indicating that TEs belonging to chimeric transcripts may be recent, polymorphic insertions. ChimeraTE is the first pipeline able to automatically uncover chimeric transcripts without a reference genome, consisting of two running Modes that can be used as a tool to investigate the contribution of TEs to transcriptome plasticity.


Unemployment in women with psychosocial disabilities during the COVID-19 pandemic: Lessons from Tana River County, Kenya.
In low-income settings, the informal economy is a practical alternative to work and employment for persons with disabilities. However, the COVID-19 pandemic negatively affected the informal economy. This study aimed to explore the experiences of women with psychosocial disabilities in Kenya during the pandemic. We found that the pandemic worsened their experiences of work and employment, and they did not receive any social welfare or support from the government. Our findings suggest that pandemic management must adopt inclusive and context-sensitive approaches that support persons with psychosocial disabilities. Social welfare and protection for persons with disabilities are relevant for socio-economic empowerment and inclusion.


Mycophenolate mofetil in animal models of autoimmune disease.
Mycofenolate mofetil (MMF-Cellcept) is an immunomodulatory drug utilized extensively in transplant medicine. The efficacy of regimes including Cellcept in preventing allograft rejection, and in the treatment of rejection, is now firmly established. The immunosuppressive actions of this drug enabled the investigation for the beneficial effects in autoimmune diseases. We review the evidence for the contribution of MMF in autoimmunity in animal models of systemic lupus erythematosus (SLE), mercury induced autoimmune glomerulonephritis, diabetes mellitus, experimental autoimmune uveoretinitis, and experimental allergic encephalitis. MMF has an influence on the T and B cell pathways. It is immunosuppressive and anti-inflammatory.


Pseudomonas aeruginosa exotoxin A induces pulmonary endothelial cytotoxicity: protection by dibutyryl-cAMP.
In pseudomonal septicaemia, serum levels of antibody to exotoxin A have been demonstrated to be an important independent predictor of survival. Previously, we have demonstrated that exotoxin A directly injures pulmonary endothelial cells, and that dibutyryl-cyclic adenosine monophosphate (Db-cAMP) can attenuate this injury. The object of this study was to examine the mechanisms of this pulmonary endothelial cell injury and the mechanism of Db-cAMP protection. The effects of differing duration of exposure to exotoxin A and a reduction in temperature on endothelial cell injury were examined. In addition, the effect of post-treatment with Db-cAMP on exotoxin A-induced endothelial cell injury was studied. A brief, 5 min, exposure to exotoxin resulted in maximum injury comparable to that produced by 18 h exposure. This injury did not occur at low temperatures, which would inhibit receptor-mediated endocytosis. Db-cAMP protected endothelial cells, even when added up to one hour after exotoxin exposure. These results suggest that, in this model, exotoxin A-induced injury of endothelial cells is receptor-mediated. Furthermore, this injury may be attenuated even after exotoxin A internalization has taken place.


Balancing work and family: A theoretical explanation and longitudinal examination of its relation to spillover and role functioning.
We take a temporally dynamic perspective to present a model that explains the relations among work-family spillover (conflict and enrichment), work-family balance, and role satisfaction and performance over time. We posit that these relationships differ for two primary conceptualizations, balance satisfaction and effectiveness. We collect data using two samples, each with three time points. In Study 1 (N = 681), we test our hypotheses for balance satisfaction. Cross-lagged analyses indicated that bidirectional enrichment predicted subsequent job and family satisfaction, and in turn, balance satisfaction. Thus, enrichment appears to primarily initiate the balance satisfaction process as it unfolds over time. Contrary to common theoretical and practical assumptions, role satisfaction seems to drive balance satisfaction rather than the other way around. In Study 2 (N = 493), we test our hypotheses for balance satisfaction and balance effectiveness. Cross-lagged analyses indicated that conflict primarily initiated the balance effectiveness process where role performance and balance effectiveness operated in feedback cycles of mutual influence over time. Posthoc model tests are consistent with Study 1 in that work-to-family enrichment predicted job satisfaction and in turn, balance satisfaction. Collectively, these studies suggest that the processes involving balance satisfaction versus balance effectiveness have different primary originating factors (enrichment or conflict, respectfully) and different temporal sequencing with role satisfaction and performance (unidirectional vs. reciprocal, respectively), warranting distinct theoretical explanations. This program of research represents a comprehensive, theoretical explanation and temporal examination of work-family balance, setting the stage for a new phase of research. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Fast, bedside diagnosis of toxic epidermal necrolysis using ex vivo confocal laser scanning microscopy: A retrospective study.
Toxic epidermal necrolysis (TEN) is a severe life-threatening drug eruption with rapid evolution. A fast histologic differentiation between TEN and clinically similarly looking staphylococcal scalded skin syndrome is of vital importance for relevant treatment decision. The recently developed ex vivo confocal laser scanning microscopy (CLSM) offers innovative and extremely fast histological visualization of fresh tissue specimens. To assess the diagnostic efficacy of ex vivo CLSM in comparison with standard histopathology for TEN. We performed side-by-side comparison of TEN specimens analysed with ex vivo CLSM and haematoxylin and eosin staining. Analysis focused on typical histopathological features of TEN, including epidermal cleavage in the basal layer and confluent epidermal necrosis. We retrospectively assessed the diagnostic performance of ex vivo CLSM for TEN in clinically confirmed cases. We report substantial agreement between ex vivo CLSM and classical histology for the detection of subepidermal cleavage and confluent epidermal necrosis. When considering full-thickness epidermal loss, epidermal cleavage in the basal layer showed the highest diagnostic performance, reaching 87.5% sensitivity and 100% specificity. Based on our data, ex vivo CSLM appears as a rapid, resource-optimizing, and reliable approach for morphological TEN emergency screening on fresh skin samples.


Low dosages of vitamin A may cause a decrease in the total neuron number of fetal hippocampal rat cells.
We investigated the effect of low, medium and high doses of oral vitamin A, on the number of fetal hippocampal neurons. High doses of vitamin A during pregnancy may cause embryonic malformations. There are reports about dosages that don't cause macroscopic malformations, but may cause mental and behavioral disorders. Still, quantitative morphological studies explaining this topic are lacking. We administered oral vitamin A to pregnant rats on the 10th-12th days of pregnancy at doses of 10000, 20000, 30000, 40000, 50000, 100000 and 200000 IU/kg. We collected the fetuses on the 19th day and removed their brains. After staining with cresyl violet and immunolabeling with Tunel and Ki67 antibody, we examined the hippocampi with stereological methods. Vitamin A decreased hippocampal neuron numbers beginning from 20000 IU/kg. While the number of Ki67 positive cells increased with the dosage, the increase of apoptotic cells begun at the dose of 50000 IU/kg. Our study demonstrates that vitamin A, beginning from the dosage of 20000 IU/kg, is decreasing the total hippocampal neuron numbers during the critical period of embryonic brain development and that apoptosis may not be the only factor in this outcome (Tab. 1, Fig. 3, Ref. 27).


Physiology of Ex Planta Nitrogenase Activity in Rhizobium japonicum.
Thirty-nine wild-type strains of Rhizobium japonicum have been studied for their ability to synthesize nitrogenase ex planta in defined liquid media under microaerobic conditions. Twenty-one produced more than trace amounts of acetylene reduction activity, but only a few of these yielded high activity. The oxygen response curves were similar for most of the nitrogenase-positive strains. The strains derepressible for activity had several phenotypic characteristics different from non-derepressible strains. These included slower growth and lower oxygen consumption under microaerobic conditions and lower extracellular polysaccharide production. Extracellular polysaccharide production during growth on gluconate in every nitrogenase-positive strain assayed was lower under both aerobic and microaerobic conditions than the non-derepressible strains. These phenotypic characteristics may be representative of a genotype of a subspecies of R. japonicum. These studies were done in part to enlarge the base number of strains available for studies on the physiology, biochemistry, and genetics of nitrogen fixation.


Role of MRI in diagnosis and management of idiopathic inflammatory myopathies.
The idiopathic inflammatory myopathies (IIM) represent a heterogeneous group of muscle diseases. Given their potential amenability to immunosuppressive treatment, the early diagnosis of the IIM subtype and the exclusion of "myositis mimics" are of great importance. MRI can visualize muscle edema, fatty replacement, atrophy, subcutaneous pathology, and fasciitis. MRI furthermore provides information about the distribution of muscle involvement and help in guiding of muscle biopsy. This review discusses significant MRI developments aimed at improving the diagnostic sensitivity and specificity, and at assessing IIM activity and damage. The manuscript also highlights techniques such as magnetic resonance spectroscopy, blood oxygenation level-dependent MRI, and diffusion-weighted MRI and T2 mapping which may in the future allow to better characterize vascular involvement, metabolic reserves, dynamic muscle recruitment, and treatment outcome.


Measurements of serum non-ceruloplasmin copper by a direct fluorescent method specific to Cu(II).
Meta-analyses indicated the breakdown of copper homeostasis in the sporadic form of Alzheimer's disease (AD), comprising copper decreases within the brain and copper increases in the blood and the pool not bound to ceruloplasmin (non-Cp Cu, also known in the literature as "free" copper). The calculated non-Cp Cu (Walshe's) index has many limitations. A direct fluorescent method for non-Cp Cu detection has been developed and data are presented herein. The study included samples from 147 healthy subjects, 36 stable mild cognitive impairment (MCI) and 89 AD patients, who were tested for non-Cp Cu through the direct method, total serum copper, ceruloplasmin concentration and o-dianisidine ceruloplasmin activity. The indirect non-Cp Cu Walshe's index was also calculated. The direct method was linear (0.9-5.9 μM), precise (within-laboratory coefficient variation of 9.7% for low and 7.1% for high measurements), and had a good recovery. A reference interval (0-1.9 μM) was determined parametrically in 147 healthy controls (27-84 years old). The variation of non-Cp Cu was evaluated according to age and sex. Non-Cp Cu was 1.5 times higher in AD patients (regarding the upper value of the reference interval) than in healthy controls. Healthy, MCI and AD subjects were differentiated through the direct non-Cp Cu method [areas under the curve (AUC)=0.755]. Considering a 95% specificity and a 1.91 μmol/L cut-off, the sensitivity was 48.3% (confidence interval 95%: 38%-58%). The likelihood ratio (LR) was 9.94 for positive test results (LR+) and 0.54 for negative test result (LR-). The direct fluorescent test reliably and accurately measures non-Cp Cu, thereby determining the probability of having AD.


Malignant gliomas display altered pH regulation by NHE1 compared with nontransformed astrocytes.
Malignant gliomas exhibit alkaline intracellular pH (pH(i)) and acidic extracellular pH (pH(e)) compared with nontransformed astrocytes, despite increased metabolic H(+) production. The acidic pH(e) limits the availability of HCO(-)(3), thereby reducing both passive and dynamic HCO(-)(3)-dependent buffering. This implies that gliomas are dependent upon dynamic HCO(-)(3)-independent H(+) buffering pathways such as the type 1 Na(+)/H(+) exchanger (NHE1). In this study, four rapidly proliferating gliomas exhibited significantly more alkaline steady-state pH(i) (pH(i) = 7.31-7.48) than normal astrocytes (pH(i) = 6.98), and increased rates of recovery from acidification, under nominally CO(2)/HCO(-)(3)-free conditions. Inhibition of NHE1 in the absence of CO(2)/HCO(-)(3) resulted in pronounced acidification of gliomas, whereas normal astrocytes were unaffected. When suspended in CO(2)/HCO(-)(3) medium astrocyte pH(i) increased, yet glioma pH(i) unexpectedly acidified, suggesting the presence of an HCO(-)(3)-dependent acid loading pathway. Nucleotide sequencing of NHE1 cDNA from the gliomas demonstrated that genetic alterations were not responsible for this altered NHE1 function. The data suggest that NHE1 activity is significantly elevated in gliomas and may provide a useful target for the development of tumor-selective therapies.


CKD and poverty: a growing global challenge.
Approximately 1.2 billion individuals worldwide live in extreme poverty (< $1/d), and 2.7 billion live in moderate poverty (< $2/d). Poverty is most prevalent in developing countries, but does not spare richer economies, where huge income discrepancies have been reported. Poverty is a major health care marker affecting a number of chronic, communicable, and noncommunicable diseases. Poverty and social deprivation are known to affect the predisposition, diagnosis, and management of chronic diseases; they directly impact on the prevalence of such conditions as obesity, diabetes, and hypertension. Also, growing evidence links poverty to chronic kidney disease (CKD). This may be caused by a direct impact of poverty on CKD or indirectly through the increased health care burden linked to poverty-associated diabetes and hypertension. Furthermore, data have shown that the poor and socially deprived have a greater prevalence of end-stage renal disease. Access to renal care, dialysis, and transplantation may also be affected by social deprivation. Overall, poverty and social deprivation are emerging as major risk markers for CKD in both developing and developed countries. Their impact on CKD warrants careful analysis because it may confound the interpretation of CKD risk factors within communities. This review therefore aims to look at the evidence linking poverty to CKD and its major risk factors, namely, diabetes and hypertension.


[Toxoplasmosis and mental diseases].
One hundred mentally handicapped patients admitted to the 10 de octubre clinical-surgical teaching hospital in the Havana city, 100 patients admitted to the day hospital of such institution and 100 normal subjects were studied. The first two groups had different psychiatric diseases. They underwent toxoplasmin tests. The percentage of reactors increased from 30% among healthy subjects to 45,9% among the those attending the day hospital and to 60% among those admitted to wards; this is statistically significative. It is concluded that the higher the severity of the psychiatric disease, the poorer the hygienic habits and the higher the contact with the parasite phocuses.


Birth weight, season of birth and postnatal growth do not predict levels of systemic inflammation in Gambian adults.
Studies testing whether systemic inflammation might lie on the causal pathway between aberrant fetal and post-natal growth patterns and later cardiovascular disease have been inconclusive, possibly due to the use of single markers of unknown predictive value. We used repeated measures of a comprehensive set of inflammatory markers to investigate the relationship between early life measures and systemic inflammation in an African population. Individuals born in three rural villages in The Gambia, and for whom early life measurements were recorded, were traced (n = 320). Fasting levels of eight inflammatory markers (C-reactive protein, serum amyloid A, orosomucoid, fibrinogen, α 1-antichymotrypsin, sialic acid, interleukin-6 and neopterin) were measured, and potential confounding factors recorded. The association between early life measurements and systemic inflammation was assessed using regression analysis. Levels of most markers were unrelated to early growth patterns. In analyses adjusted for age and sex, more rapid growth between birth and 3 months of age was associated with higher levels of fibrinogen, orosomucoid, and sialic acid. These relationships persisted after further adjustment for body mass index but after full adjustment only the association with fibrinogen remained. This study provides little evidence that size at birth or growth in early infancy determine levels of inflammatory markers in young Gambian adults.