Impact of select risk factors on treatment outcome in adults with candidemia.
Studies examining relationships between patient-related factors and treatment outcome in patients with candidemia are limited and often based on all-cause mortality. Our purpose was to examine the impact of concurrent renal replacement therapy (RRT) and other pre-specified factors on treatment outcome among adults with candidemia. This Institutional Review Board (IRB)-approved, single-center, case-cohort study included patients over 18 years of age admitted to Duke University Hospital between Jun 1, 2013 and Jun 1, 2017 with a blood culture positive for Candida spp. Treatment-, patient-, and disease-specific data were collected, and outcome (success/failure) determined 90 days after the index culture. An odds ratio (OR) and 95% confidence interval (95%CI) were calculated for the following during therapy: receipt of RRT, fluconazole monotherapy regimen, intensive care unit (ICU) stay, and neutropenia. Among the 112 encounters (from 110 unique patients) included, treatment failure occurred in 8/112 (7.1%). Demographics were comparable between outcome groups. Among 12 patients receiving concomitant RRT, only 1 patient failed therapy. With regard to treatment failure, no significant differences were observed with RRT (OR, 1.21; 95%CI, 0.14 - 10.75), fluconazole monotherapy regimen (OR, 1.59; 95%CI, 0.3-8.27), ICU stay (OR, 1.43; 95%CI, 0.32-6.29), and neutropenia (0 treatment failures). Treatment failure, receipt of concomitant RRT, and neutropenia were infrequent in patients undergoing treatment for candidemia. In our cohort, exposure to RRT, a fluconazole monotherapy regimen, ICU stay, or neutropenia during treatment did not impact treatment outcome.


A Pictorial Review of Radiologic Findings of Foreign Bodies in the Thorax.
Thoracic foreign bodies (FBs) are serious and relatively frequent in emergency departments. Thoracic FBs may occur in association with aspiration, ingestion, trauma, or iatrogenic causes. Imaging plays an important role in the identification of FBs and their dimensions, structures, and locations, before the initiation of interventional treatment. To guide proper clinical management, radiologists should be aware of the radiologic presentations and the consequences of thoracic FBs. In this pictorial essay, we reviewed the optimal imaging settings to identify FBs in the thorax, classified thoracic FBs into four types according to their etiology, and reviewed the characteristic imaging features and the possible complications.


Inactivation of thyA in Staphylococcus aureus attenuates virulence and has a strong impact on metabolism and virulence gene expression.
Staphylococcus aureus thymidine-dependent small-colony variants (TD-SCVs) are frequently isolated from patients with chronic S. aureus infections after long-term treatment with trimethoprim-sulfamethoxazole (TMP-SMX). While it has been shown that TD-SCVs were associated with mutations in thymidylate synthase (TS; thyA), the impact of such mutations on protein function is lacking. In this study, we showed that mutations in thyA were leading to inactivity of TS proteins, and TS inactivity led to tremendous impact on S. aureus physiology and virulence. Whole DNA microarray analysis of the constructed ΔthyA mutant identified severe alterations compared to the wild type. Important virulence regulators (agr, arlRS, sarA) and major virulence determinants (hla, hlb, sspAB, and geh) were downregulated, while genes important for colonization (fnbA, fnbB, spa, clfB, sdrC, and sdrD) were upregulated. The expression of genes involved in pyrimidine and purine metabolism and nucleotide interconversion changed significantly. NupC was identified as a major nucleoside transporter, which supported growth of the mutant during TMP-SMX exposure by uptake of extracellular thymidine. The ΔthyA mutant was strongly attenuated in virulence models, including a Caenorhabditis elegans killing model and an acute pneumonia mouse model. This study identified inactivation of TS as the molecular basis of clinical TD-SCV and showed that thyA activity has a major role for S. aureus virulence and physiology. Importance: Thymidine-dependent small-colony variants (TD-SCVs) of Staphylococcus aureus carry mutations in the thymidylate synthase (TS) gene (thyA) responsible for de novo synthesis of thymidylate, which is essential for DNA synthesis. TD-SCVs have been isolated from patients treated for long periods with trimethoprim-sulfamethoxazole (TMP-SMX) and are associated with chronic and recurrent infections. In the era of community-associated methicillin-resistant S. aureus, the therapeutic use of TMP-SMX is increasing. Today, the emergence of TD-SCVs is still underestimated due to misidentification in the diagnostic laboratory. This study showed for the first time that mutational inactivation of TS is the molecular basis for the TD-SCV phenotype and that TS inactivation has a strong impact on S. aureus virulence and physiology. Our study helps to understand the clinical nature of TD-SCVs, which emerge frequently once patients are treated with TMP-SMX.


The diagnostic value of pericardial fluid pH determination.
A total of 51 dogs in two different protocols each had their pericardial fluid tested for pH value. The pH values were compared with the primary diagnoses to determine if pH value is an effective discriminator between benign and neoplastic causes of pericardial effusion (PE). A high correlation appears to exist.


Percutaneous abscess drainage in patients with perforated acute appendicitis: effectiveness, safety, and prediction of outcome.
The purposes of this study were to retrospectively investigate the effectiveness and safety of CT-guided percutaneous drainage in the treatment of patients with acute appendicitis complicated by perforation and to identify CT findings and procedure-related factors predictive of clinical and procedure outcome. From March 2005 through December 2008, 41 consecutively registered patients (24 men, 17 women; age range, 18-75 years) underwent CT-guided percutaneous drainage for the management of acute appendicitis complicated by perforation and abscess. Three board-certified radiologists independently reviewed preprocedure CT images. Patients were assigned to one of three risk categories on the basis of the CT findings. Success and failure of percutaneous drainage were defined on a per-patient (i.e., clinical outcome) and per-procedure (i.e., technical outcome) basis. Immediate, periprocedure, and delayed complications were recorded. The association between candidate predictive variables, including demographic characteristics, preprocedure CT findings, and procedure-related factors and clinical or technical outcome was assessed with logistic regression models. Fifty-two CT-guided procedures were performed on 41 patients. Percutaneous drainage had clinical and technical success rates of 90% (37 of 41 patients, 47 of 52 procedures) with no procedure-related complications. In seven patients (19%) clinical success required repeated drainage procedures. A large, poorly defined periappendiceal abscess and an extraluminal appendicolith on preprocedure CT images were independent predictors of clinical failure of percutaneous drainage. CT-guided percutaneous drainage is both effective and safe in the treatment of patients with acute appendicitis complicated by perforation and abscess. The clinical and technical success rates are high.


Vedolizumab-associated psoriasis: until where does gut selectivity go?
Inflammatory Bowel Disease and Psoriasis are chronic inflammatory diseases that share common genotype, clinical course, and immunological features, although its relationship is still unclear. We report a 34-year-old woman with ileal Crohn's disease diagnosed 14 years ago, with the development of extensive, exudative scalp lesions after adalimumab therapy. Biopsies from skin lesions were compatible with vulgar psoriasis. The patient reports no personal or family history of psoriasis. Due to persistence and further worsening of skin lesions, paradoxical etiology to adalimumab was presumed and the drug was stopped with complete resolution of skin lesions and intestinal disease in remission under methotrexate. Due to pregnancy-planification methotrexate was stopped and, 8 months-after, systemic steroid-therapy was introduced due to moderate-to-severe intestinal flare. Vedolizumab was started and at the second infusion patient reported hair loss with no other complaints. Twelve months after vedolizumab initiation the patient reported reappearance of the extensive scalp and peri-fistula psoriatic lesions. Topical therapy was started but unsuccessfully and given the progressive worsening of the lesions, vedolizumab was suspended, with skin improvement seen 1 month after discontinuation. There are few case-reports of vedolizumab acting as a trigger to some dermatological conditions in IBD-patients, including psoriasis. The molecular mechanism behind it isn't fully understood. We present and discuss, to our knowledge, the first case in the literature of psoriasis triggered by vedolizumab in Crohn's disease.


The host plant metabolite glucose is the precursor of diffusible signal factor (DSF) family signals in Xanthomonas campestris.
Plant pathogen Xanthomonas campestris pv. campestris produces cis-11-methyl-2-dodecenoic acid (diffusible signal factor [DSF]) as a cell-cell communication signal to regulate biofilm dispersal and virulence factor production. Previous studies have demonstrated that DSF biosynthesis is dependent on the presence of RpfF, an enoyl-coenzyme A (CoA) hydratase, but the DSF synthetic mechanism and the influence of the host plant on DSF biosynthesis are still not clear. We show here that exogenous addition of host plant juice or ethanol extract to the growth medium of X. campestris pv. campestris could significantly boost DSF family signal production. It was subsequently revealed that X. campestris pv. campestris produces not only DSF but also BDSF (cis-2-dodecenoic acid) and another novel DSF family signal, which was designated DSF-II. BDSF was originally identified in Burkholderia cenocepacia to be involved in regulation of motility, biofilm formation, and virulence in B. cenocepacia. Functional analysis suggested that DSF-II plays a role equal to that of DSF in regulation of biofilm dispersion and virulence factor production in X. campestris pv. campestris. Furthermore, chromatographic separation led to identification of glucose as a specific molecule stimulating DSF family signal biosynthesis in X. campestris pv. campestris. (13)C-labeling experiments demonstrated that glucose acts as a substrate to provide a carbon element for DSF biosynthesis. The results of this study indicate that X. campestris pv. campestris could utilize a common metabolite of the host plant to enhance DSF family signal synthesis and therefore promote virulence.


Transcription factor PREP1 induces EMT and metastasis by controlling the TGF-β-SMAD3 pathway in non-small cell lung adenocarcinoma.
Pre-B-cell leukemia homeobox (Pbx)-regulating protein-1 (Prep1) is a ubiquitous homeoprotein involved in early development, genomic stability, insulin sensitivity, and hematopoiesis. Previously we have shown that Prep1 is a haploinsufficient tumor suppressor that inhibits neoplastic transformation by competing with myeloid ecotropic integration site 1 for binding to the common heterodimeric partner Pbx1. Epithelial-mesenchymal transition (EMT) is controlled by complex networks of proinvasive transcription factors responsive to paracrine factors such as TGF-β. Here we show that, in addition to inhibiting primary tumor growth, PREP1 is a novel EMT inducer and prometastatic transcription factor. In human non-small cell lung cancer (NSCLC) cells, PREP1 overexpression is sufficient to trigger EMT, whereas PREP1 down-regulation inhibits the induction of EMT in response to TGF-β. PREP1 modulates the cellular sensitivity to TGF-β by inducing the small mothers against decapentaplegic homolog 3 (SMAD3) nuclear translocation through mechanisms dependent, at least in part, on PREP1-mediated transactivation of a regulatory element in the SMAD3 first intron. Along with the stabilization and accumulation of PBX1, PREP1 induces the expression of multiple activator protein 1 components including the proinvasive Fos-related antigen 1 (FRA-1) oncoprotein. Both FRA-1 and PBX1 are required for the mesenchymal changes triggered by PREP1 in lung tumor cells. Finally, we show that the PREP1-induced mesenchymal transformation correlates with significantly increased lung colonization by cells overexpressing PREP1. Accordingly, we have detected PREP1 accumulation in a large number of human brain metastases of various solid tumors, including NSCLC. These findings point to a novel role of the PREP1 homeoprotein in the control of the TGF-β pathway, EMT, and metastasis in NSCLC.


Efficacy of speech pathology management for chronic cough: a randomised placebo controlled trial of treatment efficacy.
Chronic cough that persists despite medical treatment may respond to speech pathology intervention, but the efficacy of such treatment has not been investigated in prospective randomised trials. The aim of this study was to determine the efficacy of a speech pathology intervention programme for chronic cough. A single blind, randomised, placebo controlled trial was conducted in 87 patients with chronic cough that persisted despite medical treatment. Patients were randomly allocated to receive either a specifically designed speech pathology intervention or a placebo intervention. Participants in both groups attended four intervention sessions with a qualified speech pathologist. Participants in the treatment group had a significant reduction in cough (8.9 to 4.6, p<0.001), breathing (7.9 to 4.7, p<0.001), voice (7.3 to 4.6, p<0.001) upper airway (8.9 to 5.9, p<0.001) symptom scores and limitation (2.3 to 1.6, p<0.001) ratings following intervention. There was also a significant reduction in breathing (6.8 to 5.6, p = 0.047), cough (7.6 to 6.3, p = 0.014), and limitation (2.3 to 2.0, p = 0.038) scores in the placebo group, but the degree of improvement was significantly less than in the treatment group (p<0.01). Clinical judgement of outcome indicated successful ratings in 88% of participants in the treatment group compared with 14% in the placebo group (p<0.001). Speech pathology is an effective management intervention for chronic cough which may be a viable alternative for patients who do not respond to medical treatment.


Time-Series Analysis of Health Care-Associated Infections in a New Hospital With All Private Rooms.
Health care-associated infections are often caused by multidrug-resistant organisms and substantially factor into hospital costs and avoidable iatrogenic harm. Although it is recommended that new facilities be built with single-room, low-acuity beds, this process is costly and evidence of reductions in health care-associated infections is weak. To examine whether single-patient rooms are associated with decreased rates of common multidrug-resistant organism transmissions and health care-associated infections. A time-series analysis comparing institution-level rates of new multidrug-resistant organism colonization and health care-associated infections before (January 1, 2013-March 31, 2015) and after (April 1, 2015-March 31, 2018) the move to the hospital with 100% single-patient rooms. In the largest hospital move in Canadian history, inpatients in an older, tertiary care, 417-bed hospital in Montréal, Canada, that consisted of mainly mixed 3- and 4-person ward-type rooms were moved to a new 350-bed facility with all private rooms. A synchronized move of all patients on April 26, 2015, to a new hospital with 100% single-patient rooms equipped with individual toilets and showers and easy access to sinks for hand washing. Rates of nosocomial vancomycin-resistant Enterococcus (VRE) and methicillin-resistant Staphylococcus aureus (MRSA) colonization, VRE and MRSA infection, and Clostridioides difficile (formerly known as Clostridium difficile) infection (CDI) per 10 000 patient-days. Compared with the 27 months before, during the 36 months after the hospital move, an immediate and sustained reduction in nosocomial VRE colonization (from 766 to 209 colonizations; incidence rate ratio [IRR], 0.25; 95% CI, 0.19-0.34) and MRSA colonization (from 129 to 112 colonizations; IRR, 0.57; 95% CI, 0.33-0.96) was noted, as well as VRE infection (from 55 to 14 infections; IRR, 0.30, 95% CI, 0.12-0.75). Rates of CDI (from 236 to 223 infections; IRR, 0.95; 95% CI, 0.51-1.76) and MRSA infection (from 27 to 37 infections; IRR, 0.89, 95% CI, 0.34-2.29) did not decrease. The move to a new hospital with exclusively single-patient rooms appeared to be associated with a sustained decrease in the rates of new MRSA and VRE colonization and VRE infection; however, the move was not associated with a reduction in CDI or MRSA infection. These findings may have important implications for the role of hospital construction in facilitating infection control.


Why Let the Dogs Out? Exploring Variables Associated with Dog Confinement and General Characteristics of the Free-ranging Owned-Dog Population in a Peri-urban Area.
Free-ranging dogs (FRDs), are a problem in several countries, with impacts on humans, domestic animals, and wildlife, although increasing evidence suggests that most FRDs are owned. Therefore, understanding dog ownership on a fine scale is critical. The main objectives of this study were to explore dog management in rural localities from central Chile focusing on modeling owner-related variables associated with dog confinement and characterize confined and FRDs populations. Interviews (170) were carried out in Paine municipality, reporting a human:dog ratio of 1.5:1, and dogs in most households (85.9%, 146/170). Thirty-seven percent (54/146) of those households did not confine their dog(s) to some degree, and 41% (196/472) of surveyed dogs were FRD. Based on multivariable logistic regression models, non-confinement was decreased by (i) negative opinion of owners toward roaming behavior of their dogs, (ii) negative opinion toward FRDs, among others. Dog confinement increased along with owners' concerns about the impacts of their dogs on others. Owned-FRDs tended to have poorer general care than confined dogs. Our findings represent a contribution to the understanding of the human dimensions behind FRDs and provide critical quantitative elements to consider when planning effective control strategies.


Potential Role and Excretion Level of Urinary Transferrin, KIM-1, RBP, MCP-1 and NGAL Markers in Diabetic Nephropathy.
Analyzing urinary biomarkers may provide better insight into pathophysiological mechanisms of diabetic kidney diseases. The study aimed to analyze the pattern of selected excreted urinary biomarkers and its correlation with albuminuria and estimated glomerular filtration rate (eGFR) in patients with type 2 diabetes. A total of 185 patients with type 2 diabetes were categorized according to KDIGO guideline based on albuminuria and eGFR. The urinary markers (transferrin, KIM-1, RBP, MCP-1 and NGAL) were measured by ELISA. The urinary markers were associated with eGFR (total protein/Cr, p=0.001; RBP/Cr, p=0.007; MCP-1/Cr, p=0.023; NGAL/Cr, p=0.011) and albuminuria (total protein/Cr, p<0.001; transferrin, p<0.001; RBP/Cr, p<0.001; MCP-1/Cr, p<0.001; NGAL/Cr, p=0.002). The urinary marker levels (total protein, RBP, MCP-1, and NGAL) are elevated with severity of kidney damage and expressed more in progressive renal impairment.


[Renin-angiotensin-aldosterone system (RAAS) and its pharmacologic modulation].
The renin-angiotensin-aldosterone system (RAAS) is a neuroendocrine complex system that regulates the modulation of salt and water homeostasis, and regulation of blood pressure. Through its multiple interactions it protects the endothelium, heart, brain and kidney. In addition, the RAAS regulates the vascular response to injury and inflammation. Chronic activation/dysregulation of the RAAS leads to hypertension and perpetuates a cascade of proinflammatory, prothrombotic and atherogenic effects associated with endorgan damage (heart, brain, kidney, endothelium). Consequently, the RAAS is an important therapeutic target in these situations. This article presents an overview of physiology, pathophysiology and pharmacologic modulation of the RAAS.


Ciliary body temperatures during cyclocryotherapy in the clinically normal dog.
Three cryosurgical units were compared on the basis of the ability of each unit to freeze the ciliary body of enucleated normal canine eyes. The coldest average temperatures reached were -13.4 C, -4.3 C, and -17.6 C. One of the units was then used to freeze the ciliary body in the left eye of 25 dogs for 5 minutes. Ciliary body temperatures were monitored every 15 s during cyclocryosurgery. In 18 eyes in situ, the coldest average temperature reached was -13.4 C. In 7 proptosed globes, the coldest average temperature reached was -13.8 C. The results were not significantly different (P less than 0.05). The effects of the cryosurgery and centesis procedure were monitored daily by direct observation and Schiotz's tonometry. Histologic changes from the procedure were evaluated at 5 minutes, 1 week, 1 month, and 6 months after surgery. Intraocular pressure returned to the normal range within 1 to 2 weeks, the anterior uveitis subsided in 7 to 10 days, and ciliary body epithelium was histologically normal within 6 months after surgery.


[Maintenance treatment of schizophrenia with paliperidone palmitate (xeplion): a naturalistic study].
The efficacy of injectable form of paliperidone palmitate was studied in a sample of 14 schizophrenic patients (6 men and 8 women, mean age 33.8 years). Paliperidone palmitate was used as maintenance therapy in patients with remission. Duration of the study was 8 weeks. The drug was administered in doses 150 mg in the first day, 100 mg in the 8th day and 75 mg after the 4th week. Twelve (85.7%) patients responded positively to the treatment. At the end of the study, the "improvement" on CGI scale was noted in 4 patients, "minimal improvement" in 5 patients and "no change" in 3 patients. There was a significant reduction on the PANSS negative subscale. Six patients had improvement in cognitive functions. Exacerbation of positive symptoms of schizophrenia was not found. It has been concluded that injectable form of paliperidone palmitate may be recommended for maintenance treatment of schizophrenia.


Role of Salmonella enterotoxin in overall virulence of the organism.
In this study, the Salmonella enterotoxin gene (stn) was mutated by marker exchange mutagenesis, and the overall virulence of the organism was evaluated. Salmonella marker exchange mutants evoked significantly less fluid secretion in mouse intestinal loops compared to that seen with wild-type S. typhimurium. Salmonella mutants were as invasive as wild-type bacteria for HeLa cells; however, their capacity to cause destruction of the intestinal mucosa was impaired, when compared with wild-type bacteria by electron microscopy. Upon oral challenge of mice, the LD(50)of the Salmonella mutants was greater than that for the wild-type bacteria. The fluid secretory potential, as well as a reduction in the LD(50)of these mutants was restored when the mutated stn gene was replaced by the native stn gene sequence. These mutations had no effect on the aerobic growth of these bacteria in minimal or complete medium; anaerobic growth was also not affected. With these studies, we demonstrated that the presence of an intact stn gene contributed significantly to the overall virulence of S. typhimurium in a murine model.


Optical Control of Donor Spin Qubits in Silicon.
We show how to achieve optical, spin-selective transitions from the ground state to excited orbital states of group-V donors (P, As, Sb, Bi) in silicon. We consider two approaches based on either resonant, far-infrared (IR) transitions of the neutral donor or resonant, near-IR excitonic transitions. For far-IR light, we calculate the dipole matrix elements between the valley-orbit and spin-orbit split states for all the goup-V donors using effective mass theory. We then calculate the maximum rate and amount of electron-nuclear spin-polarization achievable through optical pumping with circularly polarized light. We find this approach is most promising for Bi donors due to their large spin-orbit and valley-orbit interactions. Using near-IR light, spin-selective excitation is possible for all the donors by driving a two-photon Λ-transition from the ground state to higher orbitals with even parity. We show that externally applied electric fields or strain allow similar, spin-selective Λ-transition to odd-parity excited states. We anticipate these results will be useful for future spectroscopic investigations of donors, quantum control and state preparation of donor spin qubits, and for developing a coherent interface between donor spin qubits and single photons.


Structural basis for relief of autoinhibition of the Dbl homology domain of proto-oncogene Vav by tyrosine phosphorylation.
Rho-family GTPases transduce signals from receptors leading to changes in cell shape and motility, mitogenesis, and development. Proteins containing the Dbl homology (DH) domain are responsible for activating Rho GTPases by catalyzing the exchange of GDP for GTP. Receptor-initiated stimulation of Dbl protein Vav exchange activity involves tyrosine phosphorylation. We show through structure determination that the mVav1 DH domain is autoinhibited by an N-terminal extension, which lies in the GTPase interaction site. This extension contains the Tyr174 Src-family kinase recognition site, and phosphorylation or truncation of this peptide results in stimulation of GEF activity. NMR spectroscopy data show that the N-terminal peptide is released from the DH domain and becomes unstructured upon phosphorylation. Thus, tyrosine phosphorylation relieves autoinhibition by exposing the GTPase interaction surface of the DH domain, which is obligatory for Vav activation.


Causal relationships between plasma lipids and sepsis: A Mendelian randomization study.
Although observational studies have indicated that plasma lipids are associated with an increased risk of sepsis, due to confounders and reverse causality, the causal relationship remains unclear. This study was designed to assess the causal effects of plasma lipid levels on sepsis. We used a 2-sample Mendelian randomization (MR) method to evaluate the causal effect of plasma lipids on sepsis. MR analysis employs methods such as inverse variance weighted, MR-Egger regression, weighted median regression (WME), simple mode and weighted mode. The inverse variance weighted (IVW) method was predominantly utilized to assess causality. Heterogeneity was affirmed by Cochran Q test, while pleiotropy was corroborated by MR-Egger regression analysis. The robustness and reliability of the results were demonstrated through "leave-one-out" sensitivity analysis. Instrumental variables included 226 single-nucleotide polymorphisms (SNPs), comprising of 7 for triglyceride (TG), 169 for high-density lipoprotein cholesterol (HDL-C), and 50 for low-density lipoprotein cholesterol (LDL-C). The risk of sepsis appeared to increase with rising LDL-C levels, as indicated by the inverse variance weighted analysis (OR 1.11, 95% CI from0.99 to1.24, P = 0.068). However, no causality existed between LDL-C, HDL-C, TG and sepsis. Two-sample MR analysis indicated that increased LDL-C level is a risk factor for sepsis, while TG and HDL-C levels have protective effects against sepsis. However, no significant causal relationship was found between TG, HDL-C, and LDL-C levels and sepsis.


Regulation of neurotransmitter synthesis: from neuron to gene.
We are interested in the molecular mechanisms of the regulation of neurotransmitter related gene expression by neurotrophic factors and neuronal activity. Previous work has shown that conditioned medium of muscle cells induces choline acetyltransferase (CAT) activity and represses tyrosine hydroxylase, dopamine-beta-hydroxylase and aromatic L-amino acid decarboxylase (AADC) activities in cultured sympathetic neurons. Here, we show that a new muscle-derived cell line secretes two factors which induce CAT activity in spinal cord cultures; one of those is related to LIF, a CAT inducing factor for sympathetic neurons. Preliminary data are presented on the structure of the human AADC and CAT genes. Putative promoter regions have been coupled to reporter genes; transient transfection experiments will allow us to determine the promoter elements responsible for the regulation by neurotrophic factors. We also summarize the distribution of AADC-immunoreactive cells in rat and cat brain, which will be used as a reference for the study of the specificity of expression of AADC promoter in transgenic mice.


Enhanced killing effect of nanosecond pulse electric fields on PANC1 and Jurkat cell lines in the presence of Tween 80.
We investigated the effects of nanosecond pulse electric fields (nsPEFs) on Jurkat and PANC1 cells, which are human carcinoma cell lines, in the presence of Tween 80 (T80) at a concentration of 0.18% and demonstarted an enhanced killing effect. We used two biological assays to determine cell viability after exposing cells to nsPEFs in the presence of T80 and observed a significant increase in the killing effect of nsPEFs. We did not see a toxic effect of T80 when cells were exposed to surfactant alone. However, we saw a synergistic effect when cells exposed to T80 were combined with the nsPEFs. Increasing the time of exposure for up to 8 h in T80 led to a significant decrease in cell viability when nsPEFs were applied to cells compared to control cells. We also observed cell type-specific swelling in the presence of T80. We suggest that T80 acts as an adjuvant in facilitating the effects of nsPEFs on the cell membrane; however, the limitations of the viability assays were addressed. We conclude that T80 may increase the fragility of the cell membrane, which makes it more susceptible to nsPEF-mediated killing.


Recent advances in the fabrication and bio-medical applications of self-assembled dipeptide nanostructures.
Molecular self-assembly is a widespread natural phenomenon and has inspired several researchers to synthesize a compendium of nano/microstructures with widespread applications. Biomolecules like proteins, peptides and lipids are used as building blocks to fabricate various nanomaterials. Supramolecular peptide self-assembly continue to play a significant role in forming diverse nanostructures with numerous biomedical applications; however, dipeptides offer distinctive supremacy in their ability to self-assemble and produce a variety of nanostructures. Though several reviews have articulated the progress in the field of longer peptides or polymers and their self-assembling behavior, there is a paucity of reviews or literature covering the emerging field of dipeptide-based nanostructures. In this review, our goal is to present the recent advancements in dipeptide-based nanostructures with their potential applications.


Whole blood mitigates the acute coagulopathy of trauma and avoids the coagulopathy of crystalloid resuscitation.
The contributions of type and timing of fluid resuscitation to coagulopathy in trauma remain controversial. As part of a multifunctional resuscitation fluid research effort, we sought to further characterize the coagulation responses to resuscitation, specifically as compared to whole blood. We hypothesized that early whole blood administration mitigates the acute coagulopathy of trauma by avoiding the coagulopathy of CR resuscitation. Anesthetized rhesus macaques underwent polytraumatic, hemorrhagic shock, then a crossover study design resuscitation (n = 6 each) with either whole blood first (WB-1st) followed by crystalloid (CR); or CR-1st followed by WB. Resuscitation strategies were the following: WB-1st received 50% shed blood in 30minutes, followed by twice the shed blood volume (SBV) of CR over 30minutes and one times the SBV CR over 60minutes, where CR-1st received twice the SBV of CR over 30minutes, followed by 50% of shed blood in 30minutes, and one times the SBV CR over 60minutes. Blood samples were collected at baseline, end-of-shock, end-of-first and end-of-second resuscitation stages, and end-of-resuscitation for assessment (thromboelastometry, platelet aggregation, and plasmatic coagulation factors). Statistical analyses were conducted using two-way analysis of variance ANOVA with Bonferroni correction and t-tests; significance was at p < 0.05. Survival, blood loss, hemodynamics, and shock duration were equivalent between the groups. Compared to baseline, parameters measured at first and second resuscitation stage time points directly following CR infusion revealed abnormalities in thromboelastometry (clot formation time, α angle, and maximum clot firmness), platelet aggregation response (to collagen, arachidonic acid, and adenosine diphosphate), and plasmatic coagulation (prothrombin time, anti-thrombin 3, and fibrinogen), while whole blood infusion resulted in stabilization or correction of these parameters following its administration. These data suggest that in the setting of trauma and hemorrhagic shock, the coagulation alterations begin before intervention/resuscitation; however, these are significantly aggravated by CR resuscitation and could perhaps be best termed acute coagulopathy of resuscitation. Translational animal model.


Association between first-week propofol administration and long-term outcomes of critically ill mechanically ventilated patients: A retrospective cohort study.
Propofol is commonly used in ICUs, but its long-term effects have not been thoroughly studied. In vitro studies suggest it may harm mitochondrial function, potentially affecting clinical outcomes. This study aimed to investigate the association between substantial propofol sedation and clinical outcomes in critically ill patients. We conducted a single-centre cohort study of critically ill, mechanically ventilated (≥7 days) adults to compare patients who received a substantial dose of propofol (cumulative >500 mg) during the first week of ICU admission with those who did not. The primary outcome was the association between substantial propofol administration and 6-month mortality, adjusted for relevant covariates. Subanalyses were performed for administration in the early (day 1-3) and late (day 4-7) acute phases of critical illness due to the metabolic changes in this period. Secondary outcomes included tracheostomy need and duration, length of ICU and hospital stay (LOS), discharge destinations, ICU, hospital, and 3-month mortality. A total of 839 patients were enrolled, with 73.7 % receiving substantial propofol administration (substantial propofol dose group). Six-month all-cause mortality was 32.4 %. After adjusting for relevant variables, we found no statistically significant difference in 6-month mortality between both groups. There were also no significant differences in secondary outcomes. Our study suggests that substantial propofol administration during the first week of ICU stay in the least sick critically ill, mechanically ventilated adult patients is safe, with no significant associations found with 6-month mortality, ICU or hospital LOS, differences in discharge destinations or need for tracheostomy.


Presence of lateral eye lens crystallins in the median eye of the american chameleon.
With the use of immunofluorescence techniques, gamma globulin antibody specific for the crystallins of Anolis carolinensis lateral eye lens was applied to sections through the median (parapineal) eye of Anolis carolinensis. Only the median eye lens exhibited fluorescence specific for the crystallins; other structures were negative. These results indicate that the lens of the reptilean median eye shares tissue-specific antigenic determinants with the lens of the lateral eye. This suggests a possible evolutionary relation between the two structures, based on biochemical, as well as previously reported anatomical, criteria.


Vancomycin prophylaxis in paediatric patients following cardiac surgery: a retrospective evaluation of trough levels and associated variables.
Therapeutic drug monitoring during vancomycin administration is recommended. However, little information is available in case of paediatric vancomycin prophylaxis. The aim of this study was to analyse vancomycin trough levels on postoperative day (POD) 2 and 3 after paediatric cardio-surgery to assess the clinical predictors and outcomes associated with vancomycin concentrations and to evaluate whether adjustments are effective to target optimal levels. A retrospective study was conducted in paediatric patients receiving vancomycin prophylaxis after elective cardio-surgery. Adjustments were made if levels between 20 and 30 (halving subsequent dose) or ˃30 mg/l (dose withheld) were found. Vancomycin doses of the 100 examined children (3.7-6.4 years) were 12.8 (2.5), 9.4 (5.4) and 9.7 (4.5) mg/kg, on POD1, 2 and 3, respectively (P = 0.0001). The 200 vancomycin trough levels decreased from 16.9 (11.4) on POD2 to 14.6 (8.5) on POD3 (P = 0.003). Overall, 66 troughs were sub-target, 68 reached the optimal target and 66 were supra-target. On POD2 and 3, 32 and 27 dose adjustments were required, leading to a reduced number of patients with supra-target troughs. Neonates showed a higher number of supra-target levels with respect to non-neonatal patients on both POD2 (P = 0.003) and 3 (P = 0.0001). At multivariable regression analysis, vancomycin levels showed independent association with weight and creatinine levels on both POD2 and 3. Vancomycin levels correlated with ventilation days (P = 0.31, P = 0.039), but not with methicillin-resistant Staphylococcus aureus positivity (P = 0.69). Vancomycin prophylaxis in paediatric cardio-surgery requires strict therapeutic drug monitoring and several dosage adjustments. Supra-target troughs are frequent and neonatal age, weight and creatinine levels significantly affect vancomycin concentrations.


Promising effect of Rosa damascena extract on high-fat diet-induced nonalcoholic fatty liver.
NAFLD is a chronic liver disease that affects a high proportion of the world's population which causes metabolic and hepatic damages. Rosa damascena Mill is traditionally used as a dietary supplement for liver disorders. This study was carried out to determine the beneficial effect of standardized extract of R. damascena on animal model of nonalcoholic fatty liver disease (NAFLD). NAFLD was induced by high-fat diet (HFD) in Wistar rats. HFD rats showed an increase (p < 0.05) in the plasma lipid levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), and reduced the high-density lipoprotein (HDL) levels. R. damascena significantly reduced the elevation of final body weight, liver fat accumulation, TG, TC, LDL-C concentrations and hepatic enzymes (p < 0.05). Histopathological examination of hepatic tissue confirmed the therapeutic effect of R. damascena. Improvement of total antioxidant power activity, total thiol content, MPO enzyme activity, and also lipid peroxidation were also considered in treated animals (p < 0.05). HPLC analysis showed that phenolic compounds including gallic acid, quercetin and syringic acid are the main bioactive compounds of R. damascena hydroalcoholic extract. In conclusion, R. damascena dietary supplementation has a therapeutic effect in NAFLD. Improvement of oxidative stress associated damage in liver tissue is among the main pharmacological mechanisms involved in therapeutic activity of the plant.


Tapering fibers with complex shape.
We present a model which allows us to accurately simulate the fabrication process of complex-shaped tapered fibers. The range of possible profiles is only limited by the properties of the heat source used to shape the fiber. The model takes into account the motion of the heat source relative to the fiber as well as its temperature distribution. Our measurements and corresponding finite element method (FEM) simulations have shown a strong dependency of the temperature distribution along the fiber axis on the actual diameter of the fiber. The inclusion of this relation in the model proved to be crucial for the accuracy of the results. Our model has been verified experimentally by fabricating tapered fibers with a sinusoidally modulated waist. A comparison to the profile predicted by our model reveals an excellent agreement.


Inhibition of inducible nitric oxide synthase expression by a novel small molecule activator of the unfolded protein response.
The transcription of inducible nitric oxide synthase (iNOS) is activated by a network of proinflammatory signaling pathways. Here we describe the identification of a small molecule that downregulates the expression of iNOS mRNA and protein in cytokine-activated cells and suppresses nitric oxide production in vivo. Mechanistic analysis suggests that this small molecule, erstressin, also activates the unfolded protein response (UPR), a signaling pathway triggered by endoplasmic reticulum stress. Erstressin induces rapid phosphorylation of eIF2alpha and the alternative splicing of XBP-1, hallmark initiating events of the UPR. Further, erstressin activates the transcription of multiple genes involved in the UPR. These data suggest an inverse relationship between UPR activation and iNOS mRNA and protein expression under proinflammatory conditions.


Schistosomal MicroRNAs Isolated From Extracellular Vesicles in Sera of Infected Patients: A New Tool for Diagnosis and Follow-up of Human Schistosomiasis.
Schistosomiasis traditionally has been diagnosed by detecting eggs in stool or urine. However, the sensitivity of these examinations is limited, especially in travelers with a low worm burden. Serologic tests have a greater sensitivity, but their results remain positive regardless of treatment and thus cannot be used for follow-up of patients. We hypothesized that detection of worm microRNAs (miRNAs) in serum can overcome the drawbacks of the existing diagnostic methods. Twenty-six returning travelers with schistosomiasis (based on positive results of serologic tests or detection of ova) and 17 healthy controls were included in the study. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) amplification of miRNA extracted directly from 500 µL of serum had limited sensitivity and specificity. However, qRT-PCR analysis of RNA extracted from 200 μL of serum extracellular vesicles detected 4 schistosomal miRNAs; the sensitivity and specificity of the 2 highest expressed miRNAs (bantam and miR-2c-3p) were 86% and 84%, respectively. In 7 patients with posttreatment serum available for analysis, we observed outcomes ranging from a reduction in the schistosomal miRNA level to full recovery from disease. qRT-PCR of pathogen miRNAs isolated from extracellular vesicles in sera from infected individuals may provide a new tool for diagnosing schistosomiasis in patients with a low parasite burden. This assay could also be used for evaluating the outcome of therapy, as well as disease-control programs.


The efficacy of proton pump inhibitors for the treatment of asthma in adults: a meta-analysis.
Gastroesophageal reflux disease occurs frequently among patients with asthma. Therapy with proton pump inhibitors (PPIs) to improve asthma control remains controversial. We sought to evaluate the efficacy of PPIs in treatment of asthma using objective and subjective outcome measures. A literature search was undertaken using MEDLINE (1950-January 2010), PubMed (1950-January 2010), EMBASE (1980-January 2010), and Cochrane Central Register of Controlled Trials (through January 31, 2010). Randomized, placebo-controlled trials evaluating the efficacy of PPIs for treatment of asthma in adults were selected. The primary outcome of interest was morning peak expiratory flow (PEF) rate. Secondary outcomes included objective (evening PEF rate and forced expiratory volume in 1 second) and subjective (asthma symptoms score and Asthma Quality of Life Questionnaire score) measures. Influence of study characteristics on outcomes was examined by subgroup analyses and meta-regression. Eleven trials (2524 patients) met inclusion criteria. Overall, patients had a higher mean morning PEF rate after treatment with PPIs compared with placebo (mean difference, 8.68 L/min [95% confidence interval, 2.35-15.02]). No significant single large-study effect, temporal effect, or publication bias was seen. Subgroup analysis revealed a trend toward a larger improvement in morning PEF rate in studies enrolling only patients with gastroesophageal reflux disease (mean difference, 16.90 L/min [95% confidence interval, 0.85-32.95]). Analyses of secondary outcomes (asthma symptoms score, Asthma Quality of Life Questionnaire score, evening PEF rate, and forced expiratory volume in 1 second) showed no significant difference between PPIs and placebo. Proton pump inhibitor therapy in adults with asthma results in a small, statistically significant improvement in morning PEF rate. The magnitude of this improvement, however, is unlikely to be of meaningful clinical significance. There is insufficient evidence to recommend empirical use of PPIs for routine treatment of asthma.


Analysis of meiosis in intratesticular germ cells from subjects affected by classic Klinefelter's syndrome.
Azoospermic subjects affected by Klinefelter's syndrome may occasionally show the presence of intratesticular residual foci of spermatogenesis, and the retrieval of mature spermatozoa from the testis may permit fertility and paternity by means of intracytoplasmic sperm injection. Previous studies have demonstrated that these subjects show the presence of an increased incidence of hyperaploid spermatozoa. Here we analyzed, by fluorescence in situ hybridization using specific probes for chromosomes 8, X, and Y, the spermatogenic process and the meiotic progression of 47,XXY germ cells retrieved by fine needle aspiration of the testis in ten azoospermic patients affected by classic Klinefelter's syndrome. All patients had lower testicular volume, higher gonadotropins, and lower testosterone plasma levels compared with control subjects. Cytological analysis of the testicular cells retrieved by fine needle aspiration showed the presence of Sertoli cells only in eight subjects, while germ cells were observed in two patients. In each patient Sertoli cells showed a 47,XXY karyotype, and the same chromosome pattern was observed in spermatogonia and primary spermatocytes of patients presenting a residual spermatogenesis. Secondary spermatocytes, spermatids, and mature spermatozoa showed different sex chromosome patterns, reflecting their origin from 47,XXY spermatogonia. In conclusion, this study demonstrated that, in subjects affected by Klinefelter's syndrome, residual germ cells may be present in the testis and that 47,XXY spermatogonia are able to undergo and complete the spermatogenic process leading to mature spermatozoa. These data further suggest the need to evaluate the sex chromosome status of sperm from patients affected by Klinefelter's syndrome undergoing assisted reproductive techniques.


Neuroprotective effects of lithium: implications for the treatment of Alzheimer's disease and related neurodegenerative disorders.
Lithium is a well-established therapeutic option for the acute and long-term management of bipolar disorder and major depression. More recently, based on findings from translational research, lithium has also been regarded as a neuroprotective agent and a candidate drug for disease-modification in certain neurodegenerative disorders, namely, Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), and, more recently, Parkinson's disease (PD). The putative neuroprotective effects of lithium rely on the fact that it modulates several homeostatic mechanisms involved in neurotrophic response, autophagy, oxidative stress, inflammation, and mitochondrial function. Such a wide range of intracellular responses may be secondary to two key effects, that is, the inhibition of glycogen synthase kinase-3 beta (GSK-3β) and inositol monophosphatase (IMP) by lithium. In the present review, we revisit the neurobiological properties of lithium in light of the available evidence of its neurotrophic and neuroprotective properties, and discuss the rationale for its use in the treatment and prevention of neurodegenerative diseases.


Canadian Covid-19 pandemic public health mitigation measures at the province level.
The Covid-19 pandemic has prompted governments across the world to enforce a range of public health interventions. We introduce the Covid-19 Policy Response Canadian tracker (CPRCT) database that tracks and records implemented public health measures in every province and territory in Canada. The implementations are recorded on a four-level ordinal scale (0-3) for three domains, (Schools, Work, and Other), capturing differences in degree of response. The data-set allows the exploration of the effects of public health mitigation on the spread of Covid-19, as well as provides a near-real-time record in an accessible format that is useful for a diverse range of modeling and research questions.


Spangolite: an s = 1/2 maple leaf lattice antiferromagnet?
Spangolite, Cu(6)Al(SO(4))(OH)(12)Cl·3H(2)O, is a hydrated layered copper sulfate mineral. The Cu(2+) ions of each layer form a systematically depleted triangular lattice which approximates a maple leaf lattice. We present details of the crystal structure, which suggest that in spangolite this lattice actually comprises two species of edge linked trimers with different exchange parameters. However, magnetic susceptibility measurements show that despite the structural trimers, the magnetic properties are dominated by dimerization. The high temperature magnetic moment is strongly reduced below that expected for the six s = 1/2 in the unit cell.


Critical appraisal of the potential role of intensity modulated proton therapy in the hypofractionated treatment of advanced hepatocellular carcinoma.
To investigate the role of intensity modulated proton therapy (IMPT) for advanced hepatocellular carcinoma in comparison with volumetric modulated arc therapy (VMAT). An in-silico planning study was performed on 20 patients. The prescription dose was 60Gy in 6 fractions. Patients were planned with abdominal compression. IMPT plans were optimized with or without the inclusion of CT calibration (3%) and isocenter positioning (2,4,6mm) uncertainties. Plan robustness was appraised comparing rubust optimized plans vs standard plans and also in terms of the worst-case scenario. VMAT plans were optimized for 10FFF photon beams using 2 partial arcs. Target coverage was fully achieved by both VMAT and IMPT plans with a significant improvement in homogeneity (~25%) with IMPT. Integral dose was reduced of ~60% with IMPT while the conformality of the dose distributions was similar among techniques. The sparing of the organs at risk was strongly improved with IMPT although all clinical objectives were met for both techniques. The inclusion of the uncertainties in the optimization lead to some deterioration in the target dose homogeneity (from 40 to 80% worse with 4 or 6mm position uncertainty) while none of the coverage parameters or OAR objective was violated. The worst-case scenario analysis demonstrated the risk of a major target underdosage only in the case of the most extreme errors (6mm) with D98% in average ~12% lower than the threshold. IMPT with the support of abdominal compression, can be considered a viable solution also for advanced hepatocellular carcinoma patients. Great care shall be put in the minimization of the residual respiration and positioning uncertainties but the dosimetric advantage for organs at risk and the relative robustness on target coverage are promising factors.


"No One Asked Me If I'm Alright": A Mixed-Methods Study Exploring Information/Support Needs and Challenges Engaging Men Diagnosed With Male-Factor Infertility.
There is limited research exploring men's experiences of infertility, and fewer previous studies have examined what information and support men desire after being diagnosed specifically with male-factor infertility. We conducted a mixed-methods study utilizing a combined sequential, concurrent design (online survey/semi-structured interviews). Survey outcomes (N =12) were analyzed using quantitative data analysis, while qualitative survey data (N = 5) was analyzed by reflexive thematic analysis. Heterosexual men (>18 years), fluent in English, diagnosed solely with male-factor infertility/sub-fertility, who required assisted reproductive treatment within Australia in the past 5 years were recruited online and through fertility clinics Australia-wide. Most men reported that their information and support needs were only somewhat, slightly or not at all met. Preferred information sources on male infertility were a dedicated online resource, app, or fertility doctor/specialist, while support was preferred from fertility specialists and partners. Three themes were identified from the qualitative analysis about men's experiences and support needs when diagnosed with male infertility (a) Ultimate threat to masculinity; (b) Holistic care, and (c) the power of words. The information-rich data collected provided valuable insights into men's experiences of male-factor infertility and important considerations to improve recruitment for future research. A diagnosis of male-factor infertility has the potential to be deeply impactful and difficult to navigate for men. Adequate and holistic information, recognition of emotional impacts, proactive offers of support and sensitive language are needed to improve men's experiences when undergoing assisted reproductive technology.


Socio-economic determinants and inequities in coverage and timeliness of early childhood immunisation in rural Ghana.
To assess the extent of socio-economic inequity in coverage and timeliness of key childhood immunisations in Ghana. Secondary analysis of vaccination card data collected from babies born between January 2008 and January 2010 who were registered in the surveillance system supporting the ObaapaVita and Newhints Trials was carried out. 20 251 babies had 6 weeks' follow-up, 16 652 had 26 weeks' follow-up, and 5568 had 1 year's follow-up. We performed a descriptive analysis of coverage and timeliness of vaccinations by indicators for urban/rural status, wealth and educational attainment. The association of coverage with socio-economic indicators was tested using a chi-square-test and the association with timeliness using Cox regression. Overall coverage at 1 year of age was high (>95%) for Bacillus Calmette-Guérin (BCG), all three pentavalent diphtheria-pertussis-tetanus-haemophilus influenzae B-hepatitis B (DPTHH) doses and all polio doses except polio at birth (63%). Coverage against measles and yellow fever was 85%. Median delay for BCG was 1.7 weeks. For polio at birth, the median delay was 5 days; all other vaccine doses had median delays of 2-4 weeks. We found substantial health inequity across all socio-economic indicators for all vaccines in terms of timeliness, but not coverage at 1 year. For example, for the last DPTHH dose, the proportion of children delayed more than 8 weeks were 27% for urban children and 31% for rural children (P < 0.001), 21% in the wealthiest quintile and 41% in the poorest quintile (P < 0.001), and 9% in the most educated group and 39% in the least educated group (P < 0.001). However, 1-year coverage of the same dose remained above 90% for all levels of all socio-economic indicators. Ghana has substantial health inequity across urban/rural, socio-economic and educational divides. While overall coverage was high, most vaccines suffered from poor timeliness. We suggest that countries achieving high coverage should include timeliness indicators in their surveillance systems.


Turkish hysterectomy and mastectomy patients - depression, body image, sexual problems and spouse relationships.
The aim of this study was to compare hysterectomy and mastectomy patients in terms of depression, body image, sexual problems and spouse relations. The study group comprised 94 patients being treated in Ege University Radiation Oncology Clinic, Tulay Aktas Oncology Hospital, Izmir Aegean Obstetrics and Gynecology Training and Research Hospital for breast and gynecological cancer (42 patients underwent mastectomy, 52 patient underwent hysterectomy). Five scales were used in the study: Sociodemographic Data Form, Beck Depression Scale, Body Image Scale, Dyadic Adjustment Scale, Golombok Rust Sexual Functions Scale. Mastectomy patients were more depressive than hysterectomy patients (t = 2.78, p < 0.01). Body image levels of the patients were bad but there was no significant difference between the two patient groups (p > 0.05). Hysterectomy patients had more problems in terms of vaginismus (t = 2.32, p < 0.05), avoidance of sexual intercourse (t = 2.31, p < 0.05), communication (t = 2.06, p < 0.05), and frequency of sexual intercourse than mastectomy patients (t = 2.10, p < 0.05). As compared with compliance levels between patients and spouses; hysterectomy patients had more problems related to expression of emotions than mastectomy patients (t = 2.12, p < 0.05). In conclusion, body image was negative, mastectomy was associated with more depression and hysterectomy with greater sexual problems and difficulties with spouse relationships.


Complex regulatory activities associated with the histidine kinase PrrB in expression of photosynthesis genes in Rhodobacter sphaeroides 2.4.1.
Rhodobacter sphaeroides 2.4.1 synthesizes a specialized photosynthetic membrane upon reduction of the O2 tension below threshold levels. The genes prrB and prrA encode a sensor kinase and a response regulator, respectively, of a two-component regulatory system that presumably is involved in transduction of the signal(s) that monitors alterations in oxygen levels. A third gene, prrC, is also involved in this cascade of events. Previously, we described a mutant form of PrrB, namely, PrrB78 (J. M. Eraso and S. Kaplan, J. Bacteriol. 177:2695-2706, 1995), which results in aerobic expression of the photosynthetic apparatus. Here we examine three mutated forms of the prrB gene that have the potential to encode truncated polypeptides containing the N-terminal 6, 63, or 163 amino acids, respectively. The resulting mutant strains showed residual levels of the light-harvesting spectral complexes and had diminished photosynthetic growth rates at high light intensities with no discernible growth under intermediate or low light conditions. When either lacZ transcriptional fusions or direct mRNA determinations were used to monitor specific photosynthesis gene expression, all the mutant strains showed unexpectedly high levels of gene expression when compared to mutant strains affected in prrA. Conversely, when translational fusions were used to monitor photosynthesis gene expression in these mutant strains, expression of both puc and puf operons was reduced, especially puf expression. In light of these studies and those of the PrrB78 mutant, the data suggest that PrrA can be activated in situ by something other than PrrB, and it also appears that PrrB can function as a negative regulator acting through PrrA. Finally, we consider the role of the Prr regulatory system in the posttranscriptional control of photosynthesis gene expression.


Ecological impact of repeated applications of chlorpyrifos on zooplankton community in mesocosms under Mediterranean conditions.
The effects of the insecticide chlorpyrifos were studied in plankton-dominated mesocosms under Mediterranean conditions. Chlorpyrifos was applied four times at 1 week intervals at nominal concentrations of 0.033, 0.1, 0.33, and 1 microg/l simulating repeated agricultural applications. The lowest 7 days time weighted averaged concentrations (TWAC) during the 28 days exposure period were calculated using the FOCUS equation to express the no observed effect concentration (NOEC) values. At population level the lowest NOEC calculated was 0.012 microg/l (treatment concentration 0.033 microg/l). The most affected taxon was Cladocera (Daphnia group galeata) followed by Copepoda (cyclopoids and nauplii). No effects were observed on phytoplankton (chlorophyll-a biomass) at any treatment level. The smallest NOEC(community) calculated by means of multivariate techniques was 0.1 microg/l when expressed in terms of the nominal treatment level and 0.074 microg/l when based on the lowest 7 days TWA concentration during the 28 days application period. Indirect effects on zooplankton populations were observed due to shifts in competition and predation between populations. Compared with previous micro/mesocosm experiments simulating a single application exposure regime, results from our study revealed a lower threshold level for the most sensitive measurement endpoint (difference a factor of three (in terms of nominal treatment level), more severe indirect effects and longer recovery periods of the affected populations (> 13 weeks in the test systems treated with 1 microg/l). These differences could be attributed to the repeated pulse exposure scenario approach designed for our studies together with the particular climatic conditions involving our Mediterranean mesocosms (i.e., temperature, cladocerans life history, and algae blooms).


Highly esteemed science: An analysis of attitudes towards and perceived attributes of science in letters to the editor in two Dutch newspapers.
Understanding attitudes towards science is crucial to safeguard the future of science, the application of its results and the inclusivity of decision-making processes related to science and technology. Most studies focus on attributes of social groups to explain attitudes towards science. In this study, we aim to move the discussion forward by focusing on perceived attributes of science itself by analysing over 300 letters to the editor in two Dutch national newspapers. The authors of these letters express a large degree of trust in science as a source of societal progress, if research is conducted according to a specific set of rules. Yet, they believe that these rules are under attack. The interests of universities as organizations and individual academics as well as the involvement of industry and government in research are perceived as conflicting with these rules. We conclude with recommendations for further research and practice.


[Involvement of nonhypothalamic encephalic structures in the Hand-Schüller-Christian disease. Report of a case and review of the literature].
A case of Hand-Schüller-Christian disease which presented a pyramidal syndrome besides elements of the classical triad and concentric retraction of the visual field is reported. The rare involvement of non-hypothalamic encephalic structures in that disease is emphasized together with the favourable effect of corticotherapy and radiotherapy.


The genome as a developmental organ.
This paper applies the conceptual toolkit of Evolutionary Developmental Biology (evo-devo) to the evolution of the genome and the role of the genome in organism development. This challenges both the Modern Evolutionary Synthesis, the dominant view in evolutionary theory for much of the 20th century, and the typically unreflective analysis of heredity by evo-devo. First, the history of the marginalization of applying system-thinking to the genome is described. Next, the suggested framework is presented. Finally, its application to the evolution of genome modularity, the evolution of induced mutations, the junk DNA versus ENCODE debate, the role of drift in genome evolution, and the relationship between genome dynamics and symbiosis with microorganisms are briefly discussed.


Electron-paramagnetic-resonance parameters of molybdenum(V) in sulphite oxidase from chicken liver.
A study has been made of e.p.r. signals due to Mo(V) in reduced sulphite oxidase (EC 22.214.171.124) from chicken liver. Reduction by SO3(2-), or photochemically in the presence of a deazaflavin derivative, produces spectra indistinguishable from one another. Three types of spectra from the enzyme were distingusihed and shown to correspond to single chemical species, since they could be simulated at both 9 and 35 GHz by using the same parameters. These were the low-pH form of the enzyme, with gav. 1.9805, the high-pH form, with gav. 1.9681 and a phosphate complex, with gav. 1.9741. The low-H form shows interaction with a single exchangeable proton, with A(1H)av. (hyperfine coupling constant) = 0.98 mT, probably in the form of an MoOH group. Parameters of the signals are compared with those for signals from xanthine oxidase and nitrate reductase. The signal from the phosphate complex of sulphite oxidase in unique among anion complexes of Mo-containing enzymes in showing no hyperfine coupling to protons. There is no evidence for additional weakly coupled protons or nitrogen nuclei in the sulphite oxidase signals. The possibility is considered that the enzymic mechanism involves abstraction of a proton and two electrons from HSO3- by a Mo = O group in the enzyme.


Dietary Intake of Masters Athletes: A Systematic Review.
Dietary practices of masters athletes (MAs) may promote healthy ageing; however, they are poorly understood. The aims of this systematic review were to synthesise the literature on the dietary intakes of MAs and undertake comparisons between younger (35-50 years) and older (>50 years) MAs and the general population. A search was conducted across seven databases to identify relevant publications for screening and data extraction. Averages for energy intake (EI), macronutrients, and micronutrients were compared with data from the 2011-2012 Australian Health Survey (general population). Twenty-six studies (n = 2819) were included. Energy intake was higher for older (8908 kJ/d versus 7792 kJ/d) but not younger MAs (9073 kJ/d versus 8872 kJ/d) versus the general population. Younger versus older male MAs had higher energy and macronutrient intakes. Energy intake for older was comparable to younger female MAs (7819 kJ/d versus 7485 kJ/d), but older had higher protein, lower carbohydrate, and higher micronutrient intakes. Micronutrient intake was higher in MAs than the general population. Similar EIs for older MAs and younger general population may indicate potential for a higher-quality diet. Younger female MAs may restrict or misreport EI, requiring further investigation. There is a need for more comprehensive assessments of dietary intake in MAs to ascertain diet quality in relation to health.


Rho-kinase 2 is frequently overexpressed in hepatocellular carcinoma and involved in tumor invasion.
Deregulation of Rho family small guanosine triphosphatases has been implicated in human carcinogenesis. Rho-kinases are downstream effectors of Rho guanosine triphosphatases in the regulation of cytoskeletal reorganization and cell motility. However, their functions in human cancers remain elusive. In this study, we aimed to investigate the role of Rho-kinases in hepatocellular carcinoma (HCC) tumor progression and invasion. We first examined the expression of the two Rho-kinases (ROCK1 and ROCK2) in human HCC, and found that ROCK2 was frequently overexpressed in primary HCCs (22/41 [53.66%]). Clinico-pathological analysis revealed that overexpression of ROCK2 was significantly associated with the presence of tumor microsatellite formation (P = 0.005), suggesting that deregulation of ROCK2 may contribute to the intrahepatic metastasis of HCC. Consistently, we demonstrated that stable overexpression of ROCK2 significantly enhanced cell motility and invasiveness in HCC cells. Conversely, stable knockdown of ROCK2 by short hairpin RNA approach remarkably reduced HCC cell migration and invasion. Moreover, orthotopic liver xenograft models provided further support that stable knockdown of ROCK2 suppressed HCC invasion in vivo. Stable knockdown of ROCK2 in HCC cells significantly inhibited Golgi reorientation, myosin phosphatase phosphorylation, and formations of stress fibers, filopodia, and lamellipodia; these molecular and cellular events are crucial for cell motility and cancer invasion. Our results indicate that ROCK2 was overexpressed in human HCCs, and this overexpression was associated with a more aggressive biological behavior. Our findings also demonstrate that ROCK2 played a significant role in regulating cytoskeletal events and contributed to the invasion of HCC.


Relation of particle size of C polysaccharide complexes of group A streptococci to toxic effects on connective tissue.
The component of Group A streptococci which is responsible for the chronic, remittent, multinodular lesion of connective tissue is derived from the cell wall. Further evidence is given to support the essential role of the group-specific C polysaccharide in the production of this lesion. A series of particles containing the group-specific C polysaccharide was prepared, ranging in size from large cell wall fragments to the relatively small hapten. A comparison of the lesion producing capacity of the particles in this spectrum revealed that maximum toxic activity is associated with C polysaccharide complexes of intermediate size. The discussion considers colloidal properties associated with C polysaccharide complexes of a certain size, and the influence particle size has on persistence in tissue, as possible explanations of the relationship between the size of the C polysaccharide complex and its ability to produce the chronic lesion of connective tissue.


Coevolutionary chase in two-species systems with applications to mimicry.
We study a general dynamical model describing coevolution of two haploid populations with two alleles at a single locus under weak linear symmetric frequency-dependent selection. A novel and more realistic element of our modeling approach is that both species are allowed to evolve. We analyse conditions for "evolutionary chase" between two phenotypically similar species in which one species evolves to decrease its resemblance with the other species while this other species evolves to increase its resemblance with the first species. We apply our results to a series of simple population genetics models describing classical Müllerian and Batesian mimicries as well as intermediate cases. We show that one of the most important factors influencing the plausibility of non-equilibrium dynamics in systems of mimicry is the relationship between the strength of between-species and within-species interactions. This indicates that this relationship should be the focus of both experimental and theoretical work. Our results suggest that systematic studies of frequencies of different mimicry morphs through time may be very useful.


Recovery of herpes simplex type 1 from the celiac ganglion after renal transplantation.
Herpes simplex type 1 was recovered from the celiac ganglion of a renal transplant patient who had esophageal lesions consistent with herpes simplex esophagitis as well as peptic ulcers requiring operation. We speculate that the virus got to the celiac ganglion via retrograde spread from the stomach and/or duodenum. Further studies are needed to determine the frequency of HSV involvement in visceral autonomic ganglia and whether by reactivation it could lead to subsequent gastrointestinal ulceration.


Association between PON1-rs662 Gene Polymorphism and Diabetic Retinopathy in Population of the Qom, Iran.
Diabetic retinopathy is the most severe diabetic microvascular complication that causes changes in the vessel wall. One of the genes involved in this disease is PON1, which encodes paraoxanase1 protein in liver and kidney. It might regulate inflammatory and microvascular responses to the disease. The rs662 T>C is one of the single nucleotide polymorphisms of this gene that changes glutamine to arginine at position 192. In this study, 300 samples were collected, including 100 healthy and 100 diabetics without retinopathy, and 100 diabetics retinopathies were studied and their age range was from 30 to 80 years. Then 2.5 ml of blood was collected from all relevant individuals in tubes containing EDTANa2. This polymorphism was examined by tetra-ARMS PCR. Results showed that there is no significant correlation between genotypes and alleles related to PON1 and Diabetes (CC genotype: p=0.609; C allele: p=0.228). On the other hand, an association was observed between PON1 and diabetic retinopathy (CT+CC genotype: p<0.001; CT allele: p<0.001). Considering that the Polyphen database examined the changes caused by replacing the amino acid arginine instead of glutamine at position 129 on the protein, it does not consider these changes dangerous and has introduced this polymorphism as benign. Based on the findings of this study, the rs662 locus could be considered as one of the molecular markers in future research.


Alanine racemase mutants of Mycobacterium tuberculosis require D-alanine for growth and are defective for survival in macrophages and mice.
Alanine racemase (Alr) is an essential enzyme in most bacteria; however, some species (e.g. Listeria monocytogenes) can utilize d-amino acid transaminase (Dat) to generate d-alanine, which renders Alr non-essential. In addition to the conflicting reports on gene knockout of alr in Mycobacterium smegmatis, a recent study concluded that depletion of Alr does not affect the growth of M. smegmatis. In order to get an unambiguous answer on the essentiality of Alr in Mycobacterium tuberculosis and validate it as a drug target in vitro and in vivo, we have inactivated the alr gene of M. tuberculosis and found that it was not possible to generate an alr knockout in the absence of a complementing gene copy or d-alanine in the growth medium. The growth kinetics of the alr mutant revealed that M. tuberculosis requires very low amounts of d-alanine (5-10 µg ml(-1)) for optimum growth. Survival kinetics of the mutant in the absence of d-alanine indicated that depletion of this amino acid results in rapid loss of viability. The alr mutant was found to be defective for growth in macrophages. Analysis of phenotype in mice suggested that non-availability of d-alanine in mice leads to clearance of bacteria followed by stabilization of bacterial number in lungs and spleen. Additionally, reversal of d-cycloserine inhibition in the presence of d-alanine in M. tuberculosis suggested that Alr is the primary target of d-cycloserine. Thus, Alr of M. tuberculosis is a valid drug target and inhibition of Alr alone should result in loss of viability in vitro and in vivo.


An individualized, comprehensive asthma care treatment program.
In the conclusion of a three-part series on patient-centered asthma care, the authors describe the program developed at National Jewish Center for Immunology and Respiratory Medicine. Beginning at the time of diagnosis, the interdisciplinary intervention team works in tandem with the patient and family to build confidence, skill, and motivation in self-management.


Interaction between Clostridium thermocellum endoglucanase CelD and polypeptides derived from the cellulosome-integrating protein CipA: stoichiometry and cellulolytic activity of the complexes.
Four mini-scaffoldins were constructed from modules derived from the Clostridium thermocellum cellulosome-integrating protein CipA. Cip7 and Cip6 contained one and two cohesin modules respectively. Cip14 and Cip16, also containing one and two cohesin modules respectively, were flanked by a cellulose-binding domain. Endoglucanase CelD formed stable complexes with all mini-scaffoldins. Analytical ultracentrifugation of the complexes showed that 1 mol of CelD bound per mol of Cip14, and 2 mol of CelD bound per mol of Cip16. Under the conditions used for assaying cellulase activity, 96% of CelD alone bound to Avicel. Association with Cip14 or Cip16 increased the cellulose binding of CelD to 99%, while association with Cip7 or Cip6 decreased binding to 79 and 75% respectively. The hydrolytic activity of CelD against Avicel was increased 3-fold in complexes with Cip14 and Cip16, but remained substantially the same in complexes with Cip6 and Cip7. Addition of whole CipA also enhanced the efficiency of Avicel hydrolysis by CelD. However, even at an optimal ratio of the components, CelD-CipA complexes were somewhat less active than complexes of CelD with Cip14 or Cip16. These results suggest that the synergism observed between CelD and Cip14 or Cip16 is mostly due to the presence of the cellulose-binding domain, which promotes productive binding of the enzyme.


Matches and mismatches between the global distribution of major food crops and climate suitability.
Over the course of history, humans have moved crops from their regions of origin to new locations across the world. The social, cultural and economic drivers of these movements have generated differences not only between current distributions of crops and their climatic origins, but also between crop distributions and climate suitability for their production. Although these mismatches are particularly important to inform agricultural strategies on climate change adaptation, they have, to date, not been quantified consistently at the global level. Here, we show that the relationships between the distributions of 12 major food crops and climate suitability for their yields display strong variation globally. After investigating the role of biophysical, socio-economic and historical factors, we report that high-income world regions display a better match between crop distribution and climate suitability. In addition, although crops are farmed predominantly in the same climatic range as their wild progenitors, climate suitability is not necessarily higher there, a pattern that reflects the legacy of domestication history on current crop distribution. Our results reveal how far the global distribution of major crops diverges from their climatic optima and call for greater consideration of the multiple dimensions of the crop socio-ecological niche in climate change adaptive strategies.


The migratory behavior of T blasts to contact sensitivity reactions in activelyand passively sensitized mice.
The arrival of cells from lymph nodes immunized with the contact sensitizing agents oxazolone and picryl chloride at ears challenged with these antigens was studied inmice using the technique of labelling with -51Cr. An apparent specificity of arrival was seen because the immune cells transfered contact sensitivity passively, giving rise to an inflammatory response in the ear, to which a subpopulation of cells (T blasts) was non-specifically attracted. It was also shown that there are at least two distinct populations of cells with the ability to move to inflammatory sites: the first, found in immunized lymph nodes, moves to contact sensitivity reactions in both actively and passively sensitized mice; the second, found in bone marrow and oil-induced peritoneal exudates, moves to contact sensitivity reactions in actively sensitized mice, whereas in passively sensitized mice, the arrival of these cells at contact sensitivity reaction is poor. It is suggested that the ability of T blasts to move to sites of inflammation my be useful as an assay technique for contact sensitivity reactions.


[Glucose-independent impact of the glitazones on the cardiovascular outcome].
Insulin resistance is a major intermediate link between disordered glucose metabolism and macrovascular complications. The glitazones (pioglitazone, rosiglitazone) impact directly and selectively on the key problem of insulin resistance. In the meantime, much evidence has accumulated for a vessel-protective effect of the glitazone class of substances. Currently, major endpoint studies are investigating the extent to which, and under what conditions, a glitazone actually does influence the cardiovascular morbidity and mortality. The first results from the PROactive study--the design of which, together with the data at study begin have already been published--are expected to become available by the end of 2005.


[Collagen-polysaccharide gel as a model of intercellular substance of the connective tissue].
Gel samples forming at 37 degrees C in the solutions containing tropocollagen and various polysaccharides were examined by electron microscopy. Contracting gel clots formed in the solutions containing chondroitin sulfate, proteoglycine from the tracheal cartilage, gum arabic. Electron microscopy showed such clots to be permeated with collagen fibrillae with transverse striations and a period of 640 A. An association between the density of the forming gel and the nature of the polysaccharide component is discussed. Gel forming in the solutions containing tropocollagen and various polysaccharides is regarded as a model of the connective tissue intercellular substance.


Response to propranolol and diazepam in somatic and psychic anxiety.
A total of 12 chronically anxious psychiatric outpatients were treated with racemic propranolol (Inderal), diazepam (Valium), and placebo for one week each, using a balanced cross-over experimental design. Six patients had predominantly somatic anxiety, complaining mostly of bodily symptoms, and six had mainly psychic anxiety, complaining primarily of psychological symptoms. Clinical ratings of anxiety were made by patient and psychiatrist after each treatment. Though diazepam was in general more effective than propranolol or placebo in relieving anxiety, propranolol was more effective than placebo in patients with somatic anxiety but not in those with psychic anxiety. We suggest that propranolol should be reserved for patients whose anxiety symptoms are mainly somatic.


Phenethyl alcohol as a suppressor of the envA phenotype associated with the envA gene in Escherichia coli K-12.
In Escherichia coli K-12 the envA gene was previously shown to mediate chain formation and a decreased tolerance to several antibacterial agents. Phenethyl alcohol at low concentrations has now been found to increase the tolerance to actinomycin D, ampicillin, rifampin, and gentian violet in strains containing envA. The increased tolerance to gentian violet was correlated to a decreased uptake of the dye. A phenotype suppression of chain formation and colony morphology in envA mutants was also obtained. Except for an increase in palmitic acid, chemical analysis revealed no differences between an envA and its wild-type strain in the lipopolysaccharide part of the envelope. However, a decrease in the amount of phosphatidylglycerol and a C18: 1 fatty acid was observed in the extractable lipids of a strain containing envA. Growth in the presence of phenethyl alcohol reversed the changes in fatty acid and the phospholipid composition. Phenethyl alcohol was found to cause an immediate but transient inhibition of ribonucleic acid synthesis. It is suggested that this inhibition affects the penetrability barrier of the outer cell envelope layers in strains containing envA.


Inflammatory and transcriptional roles of poly (ADP-ribose) polymerase in ventilator-induced lung injury.
Poly (ADP-ribose) polymerase (PARP) participates in inflammation by cellular necrosis and the nuclear factor-kappa-B (NF-kappaB)-dependent transcription. The purpose of this study was to examine the roles of PARP in ventilator-induced lung injury (VILI) in normal mice lung. Male C57BL/6 mice were divided into four groups: sham tracheostomized (sham), lung-protective ventilation (LPV), VILI, and VILI with PARP inhibitor PJ34 pretreatment (PJ34+VILI) groups. Mechanical ventilation (MV) settings were peak inspiratory pressure (PIP) 15 cm H2O + positive end-expiratory pressure (PEEP) 3 cm H2O + 90 breaths per minute for the LPV group and PIP 40 cm H2O + PEEP 0 cm H2O + 90 breaths per minute for the VILI and PJ34+VILI groups. After 2 hours of MV, acute lung injury (ALI) score, wet-to-dry (W/D) weight ratio, PARP activity, and dynamic compliance (CD) were recorded. Tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), myeloperoxidase (MPO) activity, and nitrite/nitrate (NOX) in the bronchoalveolar lavage fluid and NF-kappaB DNA-binding activity in tissue homogenates were measured. The VILI group showed higher ALI score, W/D weight ratio, MPO activity, NOX, and concentrations of TNF-alpha and IL-6 along with lower CD than the sham and LPV groups (P < 0.05). In the PJ34+VILI group, PJ34 pretreatment improved all histopathologic ALI, inflammatory profiles, and pulmonary dynamics (P < 0.05). NF-kappaB activity was increased in the VILI group as compared with the sham and LPV groups (P < 0.05) and was decreased in the PJ34+VILI group as compared with the VILI group (P = 0.009). Changes in all parameters were closely correlated with the PARP activity (P < 0.05). Overactivation of PARP plays an important role in the inflammatory and transcriptional pathogenesis of VILI, and PARP inhibition has potentially beneficial effects on the prevention and treatment of VILI.


Genetic alterations of RNA leukemia viruses associated with the development of spontaneous thymic leukemia in AKR/J mice.
T1-oligonucleotide fingerprinting and mapping were used to study the expression of RNA leukemia viruses in leukemic and preleukemic AKR/J mice, with techniques designed to minimize the loss or inadvertent selection of viruses in vitro before biochemical analysis. In leukemic animals, complex mixtures of ecotropic and mink-tropic viruses were expressed. Unique but similar polytropic virus-like genomes were present in each tumor isolate. In preleukemic mice, viral isolates from the thymus that were grown on NIH3T3 fibroblasts contained genomes with non-Akv polytropic virus-related oligonucleotides. This phenomenon was not evident in fingerprints of viruses from the spleen and bone marrow of the same animals. Remarkably, the non-Akv oligonucleotides located in the 3' portion of the P15E gene, the U3 noncoding region, and the 5' part of the gp70 gene were often expressed independently. Our results suggest the following. (i) Recombinant viruses can be detected in the thymuses of young preleukemic AKR mice and increase in relative abundance with age. (ii) During in vivo generation of the recombinant leukemogenic viruses, the selection of polytropic virus-related sequences in the 3' part of p15E and the U3 region and the 5' portion of gp70 occurs independently. (iii) Independent biological properties encoded in the gp70 and p15E regions of env of the recombinant viruses may mediate viral selection or leukemogenicity. (iv) The leukemogenic polytropic viruses of AKR/J mice arise via genetic recombination involving at least three endogenous viral sequences.


Modeling aging and retinal degeneration with mitochondrial DNA mutation burden.
Somatic mitochondrial DNA (mtDNA) mutation accumulation has been observed in individuals with retinal degenerative disorders. To study the effects of aging and mtDNA mutation accumulation in the retina, a Polymerase gamma (POLG) deficiency model, the POLG D257A mutator mice (PolgD257A), was used. POLG is an enzyme responsible for regulating mtDNA replication and repair. Retinas of young and older mice with this mutation were analyzed in vivo and ex vivo to provide new insights into the contribution of age-related mitochondrial dysfunction due to mtDNA damage. Optical coherence tomography (OCT) image analysis revealed a decrease in retinal and photoreceptor thickness starting at 6 months of age in mice with the POLG D257A mutation compared to wild-type (WT) mice. Electroretinography (ERG) testing showed a significant decrease in all recorded responses at 6 months of age. Sections labeled with markers of different types of retinal cells, including cones, rods, and bipolar cells, exhibited decreased labeling starting at 6 months. However, electron microscopy analysis revealed differences in retinal pigment epithelium (RPE) mitochondria morphology beginning at 3 months. Interestingly, there was no increase in oxidative stress observed in the retina or RPE of POLGD257A mice. Additionally, POLGD257A RPE exhibited an accelerated rate of autofluorescence cytoplasmic granule formation and accumulation. Mitochondrial markers displayed decreased abundance in protein lysates obtained from retina and RPE samples. These findings suggest that the accumulation of mitochondrial DNA mutations leads to impaired mitochondrial function and accelerated aging, resulting in retinal degeneration.


Adaptive Autoregressive Model for Reduction of Noise in SPECT.
This paper presents improved autoregressive modelling (AR) to reduce noise in SPECT images. An AR filter was applied to prefilter projection images and postfilter ordered subset expectation maximisation (OSEM) reconstruction images (AR-OSEM-AR method). The performance of this method was compared with filtered back projection (FBP) preceded by Butterworth filtering (BW-FBP method) and the OSEM reconstruction method followed by Butterworth filtering (OSEM-BW method). A mathematical cylinder phantom was used for the study. It consisted of hot and cold objects. The tests were performed using three simulated SPECT datasets. Image quality was assessed by means of the percentage contrast resolution (CR%) and the full width at half maximum (FWHM) of the line spread functions of the cylinders. The BW-FBP method showed the highest CR% values and the AR-OSEM-AR method gave the lowest CR% values for cold stacks. In the analysis of hot stacks, the BW-FBP method had higher CR% values than the OSEM-BW method. The BW-FBP method exhibited the lowest FWHM values for cold stacks and the AR-OSEM-AR method for hot stacks. In conclusion, the AR-OSEM-AR method is a feasible way to remove noise from SPECT images. It has good spatial resolution for hot objects.


Regulating Phase Separation Behavior of a DEEA-TETA Biphasic Solvent Using Sulfolane for Energy-Saving CO2 Capture.
Biphasic solvents containing mixed amines have a phase separation behavior and energy-efficient regeneration for CO2 capture. However, the trade-off between the CO2 absorption capacity and the volume ratio of the CO2-rich phase presents a critical challenge to the reducing potential in regeneration energy consumption. In this study, sulfolane was proposed to regulate the phase separation behavior of a N,N-diethylethanolamine (DEEA)-triethylenetetramine (TETA) biphasic absorbent by simultaneously decreasing the volume ratio and increasing the CO2 loading of the rich phase, without sacrificing the high CO2 capacity. In the DEEA-TETA-sulfolane biphasic absorbent, sulfolane acted as a phase splitter and physical activator. The replacement of a part of H2O by hydrophobic sulfolane contributed to a substantial decrease in the volume ratio of the rich phase from 83 to 39% and an increase in CO2 loading of the rich phase from 3.10 to 4.92 mol/L. The regeneration heat decreased to 1.81 GJ/t CO2, 26.4% less than DEEA-TETA, and 54.6% less than the 5 M monoethanolamine system. Moreover, by promoting the mass transfer coefficient of CO2 in DEEA-TETA-sulfolane to 1.8 times the original DEEA-TETA system, sulfolane was validated as a physical activator. Our study provides a promising strategy for regulating the phase separation behavior of biphasic solvents and enhancing the regeneration energy efficiency for CO2 capture.


Asymptomatic bronchial hyperresponsiveness in rhinitis.
Methacholine inhalation tests are used to help in the diagnosis of asthma when spirometry is normal. However, the significance of increased methacholine responsiveness in patients with rhinitis and no symptoms of asthma is not known. One possibility is that it is a false positive result; another possibility is that it indicates subclinical asthma. We investigated these possibilities in 25 patients with rhinitis, whose attending physician had not made a diagnosis of asthma, by comparing responsiveness to methacholine expressed as the provocation concentration to cause a fall in FEV1 of 20% (PC20) with responsiveness to the natural stimulus of isocapnic hyperventilation of cold air and the diurnal variation of peak flow rate. Asthma was recognized objectively by variable airflow obstruction documented by one of the latter two tests. The PC20 ranged between 4 and greater than 64 mg/ml. In 10 patients the PC20 was less than 16 mg/ml. Five of these patients had bronchoconstriction in response to hyperventilation, and a further two patients demonstrated increased variability of peak flow rates. Thus, in seven of 10 patients, increased bronchial responsiveness was confirmed by the use of two different methods, although they were asymptomatic, and the increased response to methacholine was not a false positive result. In the remaining three patients the PC20 was borderline increased (8 to 16 mg/ml). The results indicate that methacholine responsiveness in the asthmatic range in patients with rhinitis is associated with variable airflow obstruction and subclinical asthma.


InstantLabs Listeria species food safety kit. Performance tested methods 041304.
The InstantLabs Listeria Species Food SafetyKitwas validated against the International Organization for Standardization (ISO) reference method 11290-1 for the detection of Listeria monocytogenes and other Listeria species. The matrixes (stainless steel, sealed concrete, cheddar cheese, raw shrimp, and hot dogs) were inoculated with approximately 1 CFU/test portion of various Listeria species to generate fractional positives (5-15) in 20 inoculated samples. Enrichments were also fractionally inoculated with L. monocytogenes for side-by-side testing of the InstantLabs Listeria monocytogenes Food Safety Kit. Stainless steel and sealed concrete samples were validated using 4 x 4" and 1 x 1" test areas, respectively, and enriched in Buffered Listeria Enrichment Broth (BLEB) at 35 +/- 1 degrees C for 22-28 h. All food samples were tested at 25 g or 25 mL and enriched in BLEB at 35 +/- 1 degrees C for 24-28 h. All samples were confirmed using the ISO reference method, regardless of initial screen result. The InstantLabs test method performed as well as or better than the reference method for the detection of Listeria species on stainless steel, sealed concrete, cheddar cheese, raw shrimp, and hot dogs. Inclusivity and exclusivity testing revealed no false negatives and no false positives among the 80 Listeria species and 30 non-Listeria species examined. The method was shown to be robust when variations were introduced to the enrichment time, the volume for DNA extraction, and the heat block time (data not shown).


Functional studies of hephaestin in yeast: evidence for multicopper oxidase activity in the endocytic pathway.
Hephaestin is a mammalian gene that encodes a predicted multicopper oxidase required for intestinal iron export. To examine if hephaestin can act as a ferroxidase, we studied yeast strains transformed with plasmids containing both a full-length hephaestin and a hephaestin lacking a transmembrane domain. Yeast with a deletion in FET3, which encodes a cell-surface multicopper oxidase, cannot grow on low-iron media. Expression of full-length hephaestin could complement the low-iron growth phenotype of a Delta fet3 strain. Complementation of Delta fet3 cells by hephaestin required genes that encode proteins necessary for the copper loading of Fet3p, including CCC2 and GEF1. Expression of hephaestin in Delta fet3 cells led to an increase in both iron transport and oxidase activity. These results demonstrate that hephaestin is a copper-dependent protein. In contrast with Fet3p, which is found on the cell surface, hephaestin was co-localized with Pep12p-containing vesicles. Inhibition of endocytosis or deletion of both the vacuolar iron transporters ( SMF3 and FET5 / FTH1 ) prevented hephaestin from complementing the low-iron growth phenotype of Delta fet3 cells, suggesting that hephaestin is functioning within the endocytic apparatus.


Isolation of mip (microtubule-interacting protein) mutations of Aspergillus nidulans.
We identified four mutations in two previously undescribed loci involved in microtubule function in Aspergillus nidulans as extragenic suppressors of benA33, a heat-sensitive beta-tubulin mutation. Three of the four mutations map to a locus closely linked to riboB on linkage group VIII; we designated this locus mipA (for microtubule-interacting protein). We were not able to map the remaining suppressor because of chromosomal rearrangements. However, since it recombines with riboB at a significantly higher frequency than the mipA alleles, it is unlikely to be in mipA; thus, we designated it mipB1. The mip mutations are not allelic to the previously identified loci that encode alpha- and beta-tubulin, and it is likely that mipA and mipB encode previously unidentified nontubulin proteins involved in microtubule function. Each of the mip mutations suppresses the heat sensitivity conferred by benA33 and suppresses the blockage of nuclear division and movement conferred by this mutation at high temperatures. Interactions between mipA and benA are allele specific. All of the mipA mutations are cryptic in a wild-type benA background but cause cold sensitivity in combination with benA33. These mutations also confer cold sensitivity in combination with benA31 and benA32 and reduce the resistance conferred by these mutations to the antimicrotubule agent benomyl but do not suppress the heat sensitivity conferred by these alleles. Finally, the mipA alleles suppress the heat sensitivity conferred by benA11, benA17, and benA21 but do not confer cold sensitivity in combination with these alleles.