Analysis of ambulatory electrocardiograms in 14 patients who experienced sudden cardiac death during monitoring.
The Holter monitors of 14 patients (out of 58,000 Holter recordings performed between 1978 and 1984) who experienced cardiac arrest and expired during the recording period were analyzed. Tachyarrhythmic arrest patients frequently had coronary heart disease, congestive heart failure, and prolonged QTc intervals. The highest incidence of intermediately frequent premature ventricular complexes (PVCs) occurred between 15 and 6 hours prior to death. The frequency of ventricular couplets increased toward the time of arrest. The hours with greatest frequency of ventricular tachycardia (VT) were found to be the last 5 hours of life. An increasing incidence of ST-segment changes greater than 2 mm was noted throughout all of the risk periods until the third hour prior to arrest when the incidence diminished. Conversely, the incidence of lower amplitude ST-segment changes (usually elevation) increased over the final 6 hours. The mean time of death was 0228 hours +/- 5:20. In conclusion, we observed two patterns of Holter-monitored changes which usually occurred prior to death and may represent predictors of sudden death: (1) an increasing incidence of intermediately frequent isolated PVCs followed by increased ventricular couplets and runs of VT; (2) return of high amplitude ST-segment changes toward baseline. To our knowledge, the temporal relationship of the degree of ST-segment deviation to sudden death and the time of sudden death have not been reported in large studies of Holter-monitored sudden death patients.


Outcomes for balloon pulmonary valvuloplasty in adults: comparison with a concurrent pediatric cohort.
To compare procedural outcomes of transcatheter balloon pulmonary valvuloplasty (BPV) in adults to a concurrent pediatric cohort and to identify whether BPV leads to improvement in symptoms among adults. BPV has long been utilized to relieve right ventricular outflow tract (RVOT) obstruction in neonates and young children. More recently, BPV has been performed in adults, but outcome data are limited in this population. We reviewed all cases of BPV performed at Mayo Clinic's Congenital Cardiac Cath Lab from December 1993 to July 2012. Patients were excluded if they had additional congenital heart defects, other than simple left-to-right shunt lesions. 89 children and 40 adults underwent 132 BPV procedures. Among the adults, 78% were symptomatic at the time of BPV. The most common symptoms were dyspnea and fatigue. RVOT systolic gradient was 42 ± 19 mmHg (children) and 54 ± 26 mmHg (adults) prior to BPV (P = 0.002). Adults experienced a greater reduction in gradient following BPV (32 ± 21 mmHg vs. 23 ± 19 mmHg; P = 0.007). No patients exhibited "suicide right ventricle" physiology after BPV. Among adults, mean follow-up was 1.9 years (30 days-11 years). Symptoms improved in 20 of the 24 previously symptomatic adults for whom follow-up data were available. Mean NYHA score improved from 2.2 to 1.4 following BPV. At last follow-up, 52% of adults were free of any symptoms, a significant improvement from pre-BPV (P = 0.0003). Only one patient described worsening of symptoms after BPV. BPV is an effective procedure for symptomatic adults with pulmonary valve stenosis. In addition to immediate relief of RVOT gradient, adult patients may experience symptomatic improvement following BPV.


A survey of dental residents' expectations for regenerative endodontics.
The objective was to survey a group of dental residents regarding their expectations for using regenerative endodontic procedures as part of future dental treatments. After institutional review board approval, the opinions of 32 dentists who were having postgraduate residency training to become specialists in a dental school were surveyed. The survey had 40 questions about professional status, ethical beliefs, judgment, and clinical practice. It was found that 83.9% of dentists had no continuing education or training in stem cells or regenerative endodontic procedures. Results showed that 96.8% of dentists are willing to receive training to be able to provide regenerative endodontic procedures for their patients. Of the total group, 49.1% of dentists already use membranes, scaffolds, or bioactive materials to provide dental treatment. It was determined that 47.3% of dentists agree that the costs of regenerative procedures should be comparable with current treatments. It was also found that 55.1% of dentists were unsure whether regenerative procedures would be successful. Dentists are supportive of using regenerative endodontic procedures in their dental practice, and they are willing to undergo extra training and to buy new technology to provide new procedures. Nevertheless, dentists also need more evidence for the effectiveness and safety of regenerative treatments before they will be recommended for most patients.


Repressive and defensive coping styles predict resting plasma endorphin levels in the elderly.
Jamner, L.D., Schwartz, G.E., 1986, Psychosom. Med. 48, 211-223, have proposed that repressive and defensive coping are associated with greater central endogenous opioid activity. They reasoned that high-defensive (HD) participants' attenuated distress, increased pain tolerances, attenuated somatic symptomatology, and accentuated reports of positive emotions are consistent with actions of centrally active opioid peptides. The present study assessed plasma beta-endorphin (END) levels, Marlowe Crowne Social Desirability Scale (MCSD), and Taylor Manifest Anxiety Scale (TMAS) scores in men (n = 6) and women (n = 20) between the ages of 59 and 79. Contrary to predictions, HD had lower plasma endorphin levels than did low-defensive (LD) participants. Raw MCSD scores correlated negatively with endorphin levels. Findings were significant for women only, which may have been due to the small sample of men. The results are discussed as they related to the hypothesis that defensiveness involves alteration of central opioid systems.


Cardiac Lymphatic Vessels, Transport, and Healing of the Infarcted Heart.
The lymphatic vasculature plays a key role throughout the body in regulating tissue fluid homeostasis, lipid transport, and immune surveillance. Whereas it has been appreciated that the heart relies on lymphatic vessels to maintain fluid balance and that such balance must be tightly maintained to allow for normal cardiac output, it has only recently come to light that the lymphatic vasculature might serve as a therapeutic target to promote optimal healing following myocardial ischemia and infarction. We review the subject of cardiac lymphatic vessels herein and highlight studies that imply targeting of lymphatic vessel development or transport may serve as a promising avenue for future clinical application in the context of ischemic injury.


Exploration, Inference, and Prediction in Neuroscience and Biomedicine.
Recent decades have seen dramatic progress in brain research. These advances were often buttressed by probing single variables to make circumscribed discoveries, typically through null hypothesis significance testing. New ways for generating massive data fueled tension between the traditional methodology that is used to infer statistically relevant effects in carefully chosen variables, and pattern-learning algorithms that are used to identify predictive signatures by searching through abundant information. In this article we detail the antagonistic philosophies behind two quantitative approaches: certifying robust effects in understandable variables, and evaluating how accurately a built model can forecast future outcomes. We discourage choosing analytical tools via categories such as 'statistics' or 'machine learning'. Instead, to establish reproducible knowledge about the brain, we advocate prioritizing tools in view of the core motivation of each quantitative analysis: aiming towards mechanistic insight or optimizing predictive accuracy.


Repeated split sample validation to assess logistic regression and recursive partitioning: an application to the prediction of cognitive impairment.
Screening strategies play an important part in the identification and diagnosis of illness. Testing of such strategies in a clinical trial can have important implications for the treatment of such illnesses. Before the clinical trial, however, it is important to develop a practical screening/classification procedure that accurately predicts the presence of the illness in question. Recent published studies have shown a growing preference for classification tree/recursive partitioning procedures.This paper compares the application of logistic regression and recursive partitioning to a neuropsychological data set of 252 patients recruited from four Veterans Affairs Medical Centers. Logistic regression and recursive partitioning was used to predict cognitive impairment in 12 randomly selected exploratory/validation samples. We assessed the effect of sampling on variable selection and predictive accuracy.Predictive accuracy of the logistic regression and recursive partitioning procedures was comparable across the exploratory data samples but varied across the validation samples. Based on shrinkage, both classification procedures performed equally well for the prediction of cognitive impairment across the twelve samples. While logistic regression provided an estimated probability of outcome for each patient, it required several mathematical calculations to do so. However, logistic regression selected one or two less predictors than recursive partitioning with comparable predictive accuracy. Recursive partitioning, on the other hand, readily identified patient characteristics and variable interactions, was easy to interpret clinically and required no mathematical calculations. There was a high degree of overlap of the predictor variables between the two procedures.In the context of neuropsychological screening, logistic regression and recursive partitioning performed equally well and were quite stable in the selection of predictors for the identification of patients with cognitive impairment, although recursive partitioning may be easier to use in a clinical setting because it is based on a simple decision tree.


The Role of Diet and Supplements in Vitiligo Management.
Vitiligo is an autoimmune disorder that involves the interplay between oxidative stress and the immune system. Preliminary observations suggest that the presence of gluten in the diet may play a role in vitiligo development in some patients, but to date vitiligo-specific diets have not been studied. The role of oral supplements, including vitamins, minerals, and botanicals, is increasingly being investigated as adjuncts to conventional medical treatment due to their antioxidant and immunomodulatory activity. Studies suggest that many of these agents may have some efficacy as monotherapy, but more often as adjuncts to topical agents and phototherapy.


Pulsed dye laser versus long pulsed Nd:YAG laser in the treatment of angiokeratoma of Fordyce: A randomized, comparative, observer-blinded study.
Angiokeratoma of Fordyce is typically asymptomatic, blue-to-red papules with a scaly surface located on the scrotum, shaft of penis or labia majora. They can be treated with some locally destructive treatment modalities such as excision, electrocoagulation, cryotherapy and laser. To compare the effects of the pulsed dye laser versus long pulsed Nd:YAG laser in the treatment of angiokeratoma of Fordyce. Twenty tow patients with angiokeratoma of Fordyce were included in this study. All participants received three sessions of pulsed dye laser on the selected side or part of lesional area and long pulsed Nd:YAG laser on the other side or part of lesional area. Two dermatologists independently evaluated the photographs of the baseline and two-month follow-up after last session using a grade system in which treatment response was categorized into six grades. Both PDL and long pulsed Nd:YAG laser revealed statistically significant improvements in angiokeratoma of fordyce. Comparatively, there was a statistical difference between them (overall mean improvement with PDL, 61.8%, versus Nd:YAG, 77.63%; p < 0.005). Both PDL and 1064-nm Nd:YAG laser are effective and safe in the treatment of angiokeratoma of Fordyce with better response in Nd:YAG laser than pulsed dye laser.


Catecholamines in the yolk sac epithelium of the rat.
Catecholamines were found histochemically in the visceral yolk sac of the rat from embryonic day (ED) 10, i.e. before the amines become detectable in peripheral or central neurons of the fetus. Formaldehyde-induced fluorescence was confined to the apical part of the yolk sac epithelial cells. The specificity of histofluorescence has been confirmed by borohydride reduction, microspectrofluorimetry revealing an emission peak at 480 nm and administration of reserpine. The catecholamines present were identified by mass fragmentography using N,O-trifluoroacetyl derivatives. At ED 13 both dopamine and norepinephrine were present, while only dopamine was detected at ED 18 1/2. Maternal circulation or the epithelial cells themselves appear as possible sources of these catecholamines. The occurrence of amines in the yolk sac epithelium may reflect an intracellular role of these compounds, a barrier function of the epithelium or a step in a transport to the fetus where the amines might assume regulatory functions.


Involvement of cyclic AMP and its receptor protein in the sensitivity of Escherichia coli K 12 toward serine: excretion of 2-ketobutyrate, a precursor of isoleucine.
A relationship between serine-induced growth sensitivity and the cAMP-CAP complex is established. Mutants of Escherichia coli K 12 deficient either in the cya or crp gene function exhibit a resistant phenotype on serine media although they harbor a relA allele normally leading to sensitivity toward serine. The presence of a crp allele in a cya delta relA background restores the sensitivity phenotype, while the analysis of serine resistant mutants selected from a crp cya delta relA strain shows that the mutation leading to resistance is located at, or very near, the crp gene, giving a more or less Crp- phenotype. In addition crp cya delta relA strains excrete large quantities of 2-ketobutyrate when grown on glucose M63 medium. This excretion is unambiguously linked to the presence of the crp allele and is correlated with an enhanced threonine deaminase activity. Besides, the complex regulation exerted on the acetolactate synthase activities is discussed.


A preliminary report on spinal muscular atrophy lymphoblastoid cell lines: are they an appropriate tool for drug screening?
Spinal muscular atrophy (SMA) is a neurodegenerative disease of the motor neurons that results in progressive muscle weakness. It is also the leading hereditary cause of infant mortality. Homozygous loss of the survival motor neuron (SMN1) gene causes SMA, and the number of copies of the SMN2 gene modulates the severity of the disease. Increasing the expression of the SMN2 gene by pharmacological agents is one of the therapeutic approaches currently being implemented. In this preliminary study, we investigated the effect of phenylbutyrate, a histone deacetylase (HDAC) inhibitor, on SMN2 expression in two SMA type III Epstein-Barr virus (EBV)-transformed lymphoblastoid cell lines to understand the suitability of lymphoblastoid cell lines in drug screening. These cell lines are regarded as a good source as they can easily be established from the peripheral leucocytes of patients. Quantitative analysis of SMN2 mRNA was performed on established cell lines treated with various concentrations of phenylbutyrate and for a range of incubation periods using real-time polymerase chain reaction. Western blot analysis was used to determine SMN protein levels. Real-time polymerase chain reaction and Western blot analysis demonstrated that the levels of SMN2 full-length (fl-SMN2) transcripts and protein were not increased in phenylbutyrate-treated cell lines compared to non-treated controls. These results suggest that EBV-transformed lymphoblastoid cell lines are not suitable for studying the effect of certain HDAC inhibitors on SMN2 gene expression.


Schlafen 12 restricts HIV-1 latency reversal by a codon-usage dependent post-transcriptional block in CD4+ T cells.
Latency is a major barrier towards virus elimination in HIV-1-infected individuals. Yet, the mechanisms that contribute to the maintenance of HIV-1 latency are incompletely understood. Here we describe the Schlafen 12 protein (SLFN12) as an HIV-1 restriction factor that establishes a post-transcriptional block in HIV-1-infected cells and thereby inhibits HIV-1 replication and virus reactivation from latently infected cells. The inhibitory activity is dependent on the HIV-1 codon usage and on the SLFN12 RNase active sites. Within HIV-1-infected individuals, SLFN12 expression in PBMCs correlated with HIV-1 plasma viral loads and proviral loads suggesting a link with the general activation of the immune system. Using an RNA FISH-Flow HIV-1 reactivation assay, we demonstrate that SLFN12 expression is enriched in infected cells positive for HIV-1 transcripts but negative for HIV-1 proteins. Thus, codon-usage dependent translation inhibition of HIV-1 proteins participates in HIV-1 latency and can restrict the amount of virus release after latency reversal.


Gene expression patterns during enhanced periods of visual cortex plasticity.
During a critical period in its postnatal development the mammalian visual cortex displays susceptibility to experience-dependent alterations of neuronal response properties. Plasticity represents an integrated set of developmental processes controlled by a transcriptional hierarchy that coordinates the action of many genes. To illuminate the expression of these critical genes, we examined gene expression patterns of 18371 non-redundant cDNAs in the visual cortex of cats at birth, at eye opening, at the peak of the critical period of eye dominance plasticity and in the adult cat using filter-based cDNA arrays and software-based hierarchical cluster analysis. We identified a small set of genes that were selectively expressed during the peak of the critical period for plasticity. We further examined the patterns of expression of these genes by analyzing the gene expression pattern of dark-reared chronologically older animals that are known to retain this ocular dominance plasticity beyond the chronologically defined critical period. This additional cluster assessment allowed us to separate age-related changes in the patterns of gene expression from plasticity-related changes, thus identifying a subset of genes that we define as plasticity candidate genes. Those plasticity candidate genes that have previously characterized functions include participants in second messenger systems, in cell adhesion, in transmitter recycling and cytokines, among others. Comparison of cDNA array quantitation with reverse transcription-polymerase chain reaction showed almost identical expression profiles for three genes that we examined. The expression pattern of one identified gene, opioid binding cell adhesion molecule, from the cDNA array analysis, is also in agreement with immunocytochemical results. We conclude that the approach of high-density cDNA array hybridization can be used as a useful tool for examining a complex phenomenon of developmental plasticity since it is amenable to multiple developmental stage gene expression comparisons.


Something Old, Something New, Something Borrowed, Something Fused: Novel EGFR Rearrangements in Lung Adenocarcinomas.
Mutations in EGFR stand as the archetype for somatic alterations that lead to oncogene addiction and that predict for response to targeted therapies. In this issue of Cancer Discovery, Konduri and colleagues report on a pair of novel oncogenic and actionable EGFR fusion events in a series of patients with lung adenocarcinomas, casting new light on this model gene. Cancer Discov; 6(6); 574-5. ©2016 AACRSee related article by Konduri et al., p. 601.


Reduced intimal injury to canine arteries with controlled application of vessel loops.
Arterial intimal damage caused by elastic vessel loops was quantitated to determine (1) if they produce intimal injury, (2) if the injury is proportional to the force applied, and (3) if the injury is less than that produced by vascular clamps. Vessel loops were wrapped around 30 canine artery segments (5 mm in diameter) and a gram scale was connected to each to measure the force applied. The minimal occlusive force (MOF) and 150, 200, and 250 g force were applied for 30 min. For comparison, a Fogarty vascular clamp was applied at MOF for 30 min. Arteries were then reperfused for 30 min. Half was immediately harvested (group I); the remaining half was harvested at 2 weeks (group II). Scanning electron micrographs were made of each artery's intimal surface and graded from 0 (no injury) to 5 (severe intimal fracture). Mean injury grades were calculated. The MOF for vessel loops was 99 g. In group I, arteries subjected to less than 200 g force had a mean injury grade of 1.0 +/- 0.4; while those exposed to 200-250 g force had an injury grade of 3.5 +/- 0.7. The severity of injury was linearly correlated with the force applied (r = 0.72, P less than 0.009). In group II, the injury grade of 0.4 +/- 0.5 in arteries at MOF was significantly less than the injury grade of 2.8 +/- 0.5 in those at 200-250 g force, (P less than 0.0005). Injury grades of 3.7 and 3 were seen in groups I and II, respectively, when Fogarty clamps were applied. We conclude that vessel loops applied at MOF produce minimal injury. At greater occlusive forces, the injury is proportional to the force applied approaching that of a Fogarty clamp. Intimal injuries may be minimized when vessel loops are applied at MOF.


Reducing Adverse Effects During Drug Development: The Example of Lesogaberan and Paresthesia.
Lesogaberan, a γ-aminobutyric acid (GABA)B receptor agonist, was developed for the treatment of gastroesophageal reflux disease in patients with a partial response to proton pump inhibitor therapy. A high prevalence of paresthesia was observed in healthy individuals after dosing with lesogaberan in early-phase clinical trials. The aim of this review was to gain further insight into paresthesia caused by lesogaberan by summarizing the relevant preclinical and clinical data. This study was a narrative review of the literature and unpublished data. The occurrence of paresthesia may depend on the route or rate of drug administration; several studies were conducted to test this hypothesis, and formulations were developed to minimize the occurrence of paresthesia. Phase I clinical studies showed that, in healthy individuals, paresthesia occurred soon after administration of lesogaberan in a dose-dependent manner regardless of the route of administration. The occurrence of paresthesia could be decreased by fractionating the dose or reducing the rate of administration. These findings suggest that the initial rate of absorption plays an important part in the development of paresthesia. Modified-release formulations minimize the occurrence of paresthesia while retaining the anti-reflux activity of the drug, as measured by esophageal pH and the number of transient lower esophageal sphincter relaxations. The development of lesogaberan was halted because the effect on gastroesophageal reflux disease symptoms observed in Phase II studies was not considered clinically meaningful in the target patient population. Nevertheless, it is an example of successful formulation development designed to minimize the occurrence of a compound's adverse effect while retaining its pharmacodynamic action.


T-cell recovery and evidence of persistent immune activation 12 months after severe COVID-19.
T-cell lymphopenia and functional impairment is a hallmark of severe acute coronavirus disease 2019 (COVID-19). How T-cell numbers and function evolve at later timepoints after clinical recovery remains poorly investigated. We prospectively enrolled and longitudinally sampled 173 individuals with asymptomatic to critical COVID-19 and analyzed phenotypic and functional characteristics of T cells using flow cytometry, 40-parameter mass cytometry, targeted proteomics, and functional assays. The extensive T-cell lymphopenia observed particularly in patients with severe COVID-19 during acute infection had recovered 6 months after infection, which was accompanied by a normalization of functional T-cell responses to common viral antigens. We detected persisting CD4+ and CD8+ T-cell activation up to 12 months after infection, in patients with mild and severe COVID-19, as measured by increased HLA-DR and CD38 expression on these cells. Persistent T-cell activation after COVID-19 was independent of administration of a COVID-19 vaccine post-infection. Furthermore, we identified a subgroup of patients with severe COVID-19 that presented with persistently low CD8+ T-cell counts at follow-up and exhibited a distinct phenotype during acute infection consisting of a dysfunctional T-cell response and signs of excessive pro-inflammatory cytokine production. Our study suggests that T-cell numbers and function recover in most patients after COVID-19. However, we find evidence of persistent T-cell activation up to 12 months after infection and describe a subgroup of severe COVID-19 patients with persistently low CD8+ T-cell counts exhibiting a dysregulated immune response during acute infection.


Benefit of screening mammography in reducing the rate of late-stage breast cancer diagnoses (United States).
We studied the benefit of modern mammography screening in community settings, evaluating age-related differences in rates of late-stage breast cancer detection. Our multicenter population-based case-control study included 931 black and white women with incident breast cancer (American Joint Commission on Cancer Stage IIB or higher) diagnosed 1994-1998 and 4,016 randomly sampled controls never diagnosed with breast cancer. Adjusted odds ratios (ORs) estimated the relative rate of late-stage diagnosis in screened and non-screened women. Women aged 50-64 at diagnosis with at least one screening mammogram in the previous 2 years were significantly less likely to have late-stage diagnosis (OR = 0.41, 95% CI 0.33-0.52). Results for women aged 40-49 were consistent with a screening benefit, although the confidence interval marginally overlapped the null (OR = 0.81, 95% CI 0.64-1.02). Mammography screening was associated with lower rates of late-stage breast cancer among both premenopausal (OR = 0.64, 95% CI 0.50-0.81) and postmenopausal (OR = 0.44, 95% CI 0.35-0.56) women. With modern mammography in the community, rates of late-stage breast cancer diagnoses are lower in screened compared to non-screened women ages 40 and older, but age-related differences persist.


A review: extraction, phytochemicals, and biological activities of rambutan (Nephelium lappaceum L) peel extract.
Rambutan (Nephelium lappaceum L.) peels are produced during application process. Drying methods of rambutan peel, including open sun, oven, oven vacuum, and freeze-drying, have been describes in this study. The extraction technologies of dried rambutan peels were reviewed, such as maceration and hot extraction, microwave-assisted extraction, ultrasonic-assisted extraction, and supercritical fluid extraction. The phytochemicals of rambutan peel extracts were analyzed, and the purification and stability of geraniin was reviewed. Rambutan peel extracts exhibit wide bioactivities in vitro and in vivo, and these bioactivities depend chiefly on the phenolic contents and profiles in the different extracts. The safety of rambutan peel extracts was analyzed. In addition, rambutan peel extracts could be used as important components to make different products, which are potentially applied in food, medicine, and cosmetic. However, the extracts efficiency must be further increased using some emerging technologies. Furthermore, the bioactive mechanism and bioavailability of the extract in human system should be further evaluated.


Modified method of application of ethyl methanesulfonate for induction of Escherichia coli mutants.
An efficient modification of ethyl methanesulfonate mutagenic action is based on mutagenization of small volumes of cell suspensions in micro-sample tubes. This provides for a rapid and safe handling of solutions of cancerogenic mutagens. A 3-4-h exposition to 30-40 mmol/L of mutagen appeared optimal, inducing more than 20% auxotrophs or, after a simultaneous application of the penicillin method, 60% of auxotrophs. Moreover, the modification proved its value in repeated applications of ethyl methanesulfonate, resulting in an accumulation of various mutation types. Consecutive mutations were accompanied by an increase in sensitivity. Based on the distribution of nucleoids in the mutagenized population, the penicillin method was modified to allow detection of mutants segregated from cells with several nucleoids.


Gold nanoparticles (AuNPs) can rapidly deliver artificial microRNA (AmiRNA)-ATG6 to silence ATG6 expression in Arabidopsis.
We establish a fast and efficient transient silencing system that facilitates functional studies of some genes, whose knockout leads to plant lethality. In plants, the generation of loss-of-function mutants is crucial for studying gene function. Artificial microRNA (AmiRNA) technology is a more targeted and effective tool for gene silencing. Gold nanoparticles (AuNPs) can bind nucleic acids and deliver them into animal cells. Here, AuNPs are used in combination with AmiRNA technology in plants. We found that AmiRNA-autophagy-related proteins (ATG6) can be delivered to cells by AuNPs to achieve the effect of ATG6 silencing. It is worth noting that on the 10th day there is still a silencing effect. Similar to the atg5 lines, silencing of ATG6 significantly reduced plant resistance to Pseudomonas syringae pv.maculicola (Psm) ES4326/AvrRpt2. Interestingly, ATG6 silencing and ATG5 mutation in NPR1-GFP (nonexpressor of pathogenesis-related genes) lines significantly reduced plant resistance to Psm ES4326/AvrRpt2, suggesting that autophagy is also involved in NPR1-regulated plant immune responses. In summary, we establish a fast and efficient transient silencing system that facilitates functional studies of some genes, whose knockout leads to plant lethality.


Leveraging the Combined Predictive Value of Ultrasound and Laboratory Data to Reduce Radiation Exposure and Resource Utilization in Children with Suspected Appendicitis.
Previous investigation has shown that the combined predictive value of white blood cell count and ultrasound (US) findings to be superior to either alone in children with suspected appendicitis. The purpose of this study was to evaluate the impact of a diagnostic clinical pathway (DCP) leveraging the combined predictive value of these tests on computed tomography (CT) utilization and resource utilization. Retrospective cohort study comparing 8 mo of data before DCP implementation to 18 mo of data following implementation. The pathway incorporated decision-support for disposition (operative intervention, observation, or further cross-sectional imaging) based on the combined predictive value of laboratory and US data (stratifying patients into low, moderate, and high-risk groups). Study measures included CT and magnetic resonance imaging utilization, imaging-related cost, time to appendectomy, and negative appendectomy rate. Ninety-seven patients in the preintervention period were compared with 319 patients in the postintervention period. Following DCP implementation, CT utilization decreased by 86% (21% versus 3%, P < 0.001). Mean time to appendectomy decreased from 8.5 to 7.2 h (P < 0.001), and the negative appendectomy rate remained unchanged (5% versus 4%, P = 0.54). Magnetic resonance imaging utilization increased following pathway implementation (1% versus 7%, P = 0.02); however, median imaging-related cost was significantly lower in the postimplementation period ($283/case to $270/case, P = 0.002) CONCLUSIONS: In children with suspected appendicitis, implementation of a DCP leveraging the combined predictive value of white blood cell and US data was associated with a reduction in CT utilization, time to appendectomy, and imaging-related cost.


Maxillary sinus carcinoma: the only symptom was neck lymph node swelling.
Since maxillary sinus is composed of bone structure, the main symptoms of maxillary sinus carcinoma are related to the anatomical feature and the destructive lesion of the bony wall, such as cheek pain and nasal obstruction. We report a female case with undifferentiated carcinoma in the right maxillary sinus, only appearing cervical swelling which was revealed as lymph node metastasis. CT and MRI findings showed just maxillary sinusitis with minor bone destruction. However, fluorine 18-labelled deoxyglucose positron emission tomography (FDG-PET) was useful for the detection of the primary site. The patient received concomitant chemoradiotherapy, and showed a complete response both in the primary site and neck lymph nodes. She has no recurrence for 18 months after the primary therapy. The main symptoms of maxillary sinus carcinoma are related to the local progression, and known to have less cervical lymph node metastasis. However, like the present case, there is a rare case that has no symptom and organic features associated with the local mass. With the best use of advanced diagnostic technique, e.g., FDG-PET, we could diagnose and treat atypical maxillary sinus carcinoma patients.


Analysis of site-specific late potentials using a novel Holter signal-averaged electrocardiography in patients with Brugada syndrome.
The utility of late potentials on signal-averaged electrocardiography (SAECG) for risk stratification in patients with Brugada syndrome (BrS) remains controversial. Late potentials on conventional SAECG with Frank leads may be insufficiently sensitive to detect site-specific late potentials in right precordial leads. The purpose of this study was to evaluate the utility of site-specific late potentials using a novel unipolar Holter-SAECG system for risk stratification in patients with BrS. Consecutive symptomatic (n = 20) and asymptomatic (n = 21) patients with BrS who underwent investigation using conventional SAECG and a novel unipolar Holter-SAECG system were enrolled. We evaluated clinical characteristics and outcomes and compared late potentials on the 2 SAECGs between both groups and patients with and without cardiac events (CEs) (sudden cardiac death or sustained ventricular tachyarrhythmias) during the follow-up period. During mean follow-up of 76 months, 10 patients (24%) had CEs. There were no significant differences in late potentials on conventional SAECG between symptomatic and asymptomatic patients. On the Holter-SAECG system, RMS40 in lead V2 in the third intercostal space (3L-V2) at the nighttime was significantly lower in the symptomatic group than in the asymptomatic group (5.5 ± 0.8 μV and 8.2 ± 0.8 μV, respectively; P = .027). Univariate analysis of predictive values for CE showed that hazard ratios of daytime and nighttime RMS40 in lead 3L-V2 of <7.7 μV and <6.1 μV were 7.58 and 6.14, respectively. Site-specific late potentials in lead 3L-V2 measured using the novel Holter-SAECG system may be a useful marker for high-risk patients with BrS.


Can a lightbulb sestamibi SPECT accurately predict single-gland disease in sporadic primary hyperparathyroidism?
Technetium-99m sestamibi scintigraphy with single photon emission computed tomography (SPECT) is widely used to guide minimally invasive exploration in patients with sporadic primary hyperparathyroidism (SPH), although its sensitivity in multiglandular disease is limited. We examined the incidence of missed multiglandular disease and associated anatomic findings when sestamibi SPECT was positive for a single intense focus of delayed tracer uptake, termed a lightbulb scan (LBS). Prospectively entered data from 764 patients with SPH treated with initial parathyroid exploration from March 5, 2000, to December 31, 2006, were reviewed. A single radiologist performed blinded interpretation of 585 available sestamibi SPECT images, classifying 167 (28.5%) patients with a LBS. Clinical findings were compared among LBS patients with a single adenoma (true positive) and LBS patients with multiglandular disease (false negative). One hundred fifty of 167 (89.8%) LBS patients had a single adenoma and 3 (1.8%) had carcinoma. Multiglandular disease was anatomically present in 14 of 167 (8.4%) LBS patients compared with 60 of 418 (15.6%) non-LBS patients (p=0.05). Parathyroid hyperplasia occurred less frequently in LBS patients [5/167 (3%)] compared with non-LBS patients [36/418 (8.6%)], (p=0.02), while double adenomas occurred equally often in LBS patients [9/167 (5.4%)] compared with non-LBS patients [24/418 (5.7%)], (p=0.87). Double adenomas in LBS patients were more likely ipsilateral (7/9, p=0.005) and left-sided (7/7, p=0.008). LBS patients with multiglandular disease were more likely to have a history of neck irradiation, prior neck exploration, and concomitant thyroid pathology. In patients with SPH, sestamibi SPECT studies show a single bright focus of uptake in only 29% of patients. LBS findings do not exclude multiglandular disease. To avoid unacceptable rates of failure at initial parathyroid exploration, the expert surgeon should use validated adjuncts such as intraoperative PTH monitoring or four-gland exploration.


Regeneration of biomimetic hydroxyapatite on etched human enamel by anionic PAMAM template in vitro.
To repair the demineralized enamel by biomimetic method, and the effect of Poly (amido amine) (PAMAM) dendrimers on the crystallization of hydroxyapatite on etched enamel surface is investigated. PAMAM dendrimers were synthesized step by step following the classical method and modified with the carboxylic acid groups (COOH) on the surface. Demineralized human enamel samples were immersed in 10,000ppm PAMAM-COOH solution for 30min and then in calcium phosphorous solution with or without fluoride under near-clinical conditions for 20h. Other samples without PAMAM-COOH were immersed in calcium phosphorous solution as the control group. After the immersion, the micro structure, morphology and composition of the regrown crystals on the longitudinal and transversal enamel surfaces were investigated by SEM, XRD and FTIR, and the results were compared with etched enamel and intact enamel. With the PAMAM-COOH templates, well-arranged rod-like crystals were formed and they were parallel to the long axis of enamel crystals, which was more obvious on the longitudinal enamel surface. Otherwise, irregular flake-like crystals were obtained without PAMAM-COOH. Fluorapatite was not influenced by the PAMAM-COOH but its specific distribution also shown the patterns of the PAMAM-COOH temples XRD spectra showed that the main phase of the obtained crystals with PAMAM-COOH was hydroxyapatite and their morphology and structure were close to the intact enamel. Amide I band and two bands of methylene groups of PAMAM-COOH detected by FTIR demonstrated the presence of PAMAM-COOH within the biomimetic coating. It was concluded that PAMAM-COOH can play as the organic template on the demineralized enamel surface to induce the formation of HAP crystals with the same structure, orientation and mineral phase of the intact enamel in relatively short time.


A systematic review of tuberosity healing and outcomes following reverse shoulder arthroplasty for fracture according to humeral inclination of the prosthesis.
Proximal humerus fractures are common in the elderly population and are often treated with reverse shoulder arthroplasty (RSA). The purpose of this systematic review was to compare tuberosity healing and functional outcomes in patients undergoing RSA with humeral inclinations of 135°, 145°, and 155°. A systematic review was performed of RSA for proximal humerus fracture using Preferred Reporting Items for Systemic Reviews and Meta-Analyses (PRISMA) guidelines. Radiographic and functional outcome data were extracted to evaluate tuberosity healing according to humeral inclination. Analysis was also performed of healed vs. nonhealed tuberosities. A total of 873 patients in 21 studies were included in the analysis. The mean age was 77.5 years (range of 58-97) and the mean follow-up was 26.2 months. Tuberosity healing was 83% in the 135° group compared with 69% in the 145° group and 66% in the 155° group (P = .030). Postoperative abduction was highest in the 155° group (P < .001). No significant difference was found in forward flexion, external rotation, or postoperative Constant score between groups. Patients with tuberosity healing demonstrated 18° higher forward flexion (P = .008) and 16° greater external rotation (P < .001) than those with unhealed tuberosities. RSA for fracture with 135° humeral inclination is associated with higher tuberosity healing rates compared with 145° or 155°. Postoperative abduction is highest with a 155° implant, but there is no difference in in postoperative forward flexion, external rotation, or Constant score according to humeral inclination. Patients with healed tuberosities have superior postoperative forward flexion and external rotation than those with unhealed tuberosities.


Design, synthesis, and antifungal activity of inhibitors of brassilexin detoxification in the plant pathogenic fungus Leptosphaeria maculans.
Potential inhibitors of Leptosphaeria maculans mediated detoxification of the phytoalexin brassilexin were designed and synthesized based on the planar heteroaromatic structure of isothiazolo[5,4-b]indole. Screening of these compounds for inhibition of brassilexin detoxification in cultures of L. maculans indicated that 4-(2-chlorophenyl)isothiazole had the largest effect on the rate of brassilexin detoxification. However, the most antifungal compound among the potential inhibitors, isothiazolo[5,4-b]quinoline, did not appear to affect the metabolism of brassilexin noticeably, suggesting that growth inhibition is not sufficient to slow down the rate of brassilexin detoxification. Furthermore, it was determined that 4-arylisothiazoles as well as isothiazolo[5,4-b]thianaphthene displayed antifungal activity against L. maculans.


Spray dried microspheres based on chitosan: A promising new carrier for intranasal administration of polymeric antigen BLSOmp31 for prevention of ovine brucellosis.
Previous studies have demonstrated that parenteral immunization with polymeric antigen BLSOmp31 induced a strong immune response and conferred protection against Brucella ovis in rams. This work describes the development of a novel formulation strategy for the delivery of BLSOmp31 in the nasal mucosa. Chitosan microparticles were prepared by spray-drying technology processes and recombinant chimera BLSOmp31 was loaded by passive adsorption onto chitosan microspheres, which were characterized by means of the evaluation of size, zeta potential, morphology, and loading and release rate of BLSOmp31. The mucoadhesive properties of microspheres were evaluated by studying the interaction between microparticles and mucin. The antigen BLSOmp31 integrity was investigated by SDS-PAGE. The yield of production of spray-drying process was 68.95%. Microspheres had a good sphericity, 1-10 μm of particle size and had a positive charge. The loading capacity was found to be 45.19%. The initial fast release of BLSOmp31 from chitosan microparticles was 60%. The BLSOmp31 integrity was not affected by passive adsorption (ionic interaction). The amount of mucin adsorbed on the surface of CMs-BLSOmp31 was lower than on the surface of blank CMs at neutral pH. In vivo studies were carried out in rams. Intranasal immunization induced systemic and local antibodies. In conclusion, the use of BLSOmp31-loaded chitosan spray-drying microspheres offers a promising way for nasal mucosal vaccination in sheep against brucellosis.


Comparison of extraction and clean-up procedures for analysis of zearalenone in corn, rice and wheat grains by high-performance liquid chromatography with photodiode array and fluorescence detection.
The aim of this work was the optimization of some procedures usually used in the analysis of zearalenone (ZEA) in corn and other cereals by high-performance liquid chromatography (HPLC) with photodiode array and/or fluorescence detection. The comparison of five extraction solvents is presented. Three solid-phase extraction cartridges (C-18, silica, Florisil) and immuno-affinity columns were also compared to obtain the best recovery of the mycotoxin with the minimal presence of co-extractives in the chromatograms. Mixtures of methanol-1% aqueous NaCl (80.20 or 60:40 v/v) were the best extraction solvents. Florisil provided higher recovery of ZEA than C-18, and silica proved unsuitable. The immuno-affinity column was very efficient in cleaning the extracts, but its sample capacity was lower than that of SPE columns due to saturation. The mobile phase (methanol-water 80:20 v/v) gave a low retention time for ZEA (approximately 5 min), high sensitivity and acceptable separation between this mycotoxin and alpha-zearalenol. The optimized protocol is straightforward, provides high ZEA recoveries in spiked corn (mean 102.4%), has an acceptable sensitivity and has a lack of interference with fluorescence detection (detection limit 4 ng ZEA g(-1) corn). The photodiode array detector was useful, except at very low ZEA levels, to confirm the identity of the mycotoxin. The method was applied to search for ZEA accumulation in corn, wheat and rice grains inoculated with selected strains of Fusarium graminearum, F. oxysporum and method was applied to search for ZEA accumulation in corn, wheat and rice grains inoculated with selected strains of Fusarium graminearum, F. oxysporum and F. culmorum.


Detection of Helical Intermediates During Amyloid Formation by Intrinsically Disordered Polypeptides and Proteins.
Amyloid formation and aberrant protein aggregation are hallmarks of more than 30 different human diseases. The proteins that form amyloid can be divided into two structural classes: those that form compact, well-ordered, globular structures in their unaggregated state and those that are intrinsically disordered in their unaggregated states. The latter include the Aβ peptide of Alzheimer's disease, islet amyloid polypeptide (IAPP, amylin) implicated in type 2 diabetes and α-synuclein, which is linked to Parkinson's disease. Work in the last 10 years has highlighted the potential role of pre-amyloid intermediates in cytotoxicity and has focused attention on their properties. A number of intrinsically disordered proteins appear to form helical intermediates during amyloid formation. We discuss the spectroscopic methods employed to detect and characterize helical intermediates in homogenous solution and in membrane-catalyzed amyloid formation, with the emphasis on the application of circular dichroism (CD). IAPP is used as an example, but the methods are generally applicable.


Ontogeny of cholecystokinin satiety in rats.
The development of cholecystokinin satiety effects was studied in newborn rats. Weight gain during tests was the measure of intake in the sucklings, until the rats were weaned at 20 days of age. Thereafter food intake was measured directly. Intraperitoneal injection of cholecystokinin (CCK-OP) suppressed feeding in 1-7 day old rat pups. Gastric loads of 0.9% NaCl, 2.5% D-phenylalanine, and L-phenylalanine suppressed subsequent intake in 1-10 day old rats, but not in 10-20 day old rats. The early satiety effect of the intragastric loads was attributed to gastric distension. At 21-25 days of age, rats showed a suppression of intake following gastric loads of L-phenylalanine but not D-phenylalanine. This is tentative evidence that releasers of endogenous CCK do not induce satiety until immediately after weaning.


Microcalcifications in the breast detected by a color Doppler method using twinkling artifacts: some important discussions based on clinical cases and experiments with a new ultrasound modality called multidetector-ultrasonography (MD-US).
The twinkling artifact is well known as a color Doppler artifact but it is still an unresolved phenomenon [Rahmouni et al., Radiology 1996;199:269-271 ; Atan et al., Astas Urol Esp 2001;35:396-402; Kamaya et al., AJR 2003;80:215-222]. Many factors affect the appearance of the twinkling artifact, such as the surface roughness of stones creating multiple reflections and a form of intrinsic noise known as clock jitter within the Doppler circuitry of the ultrasound equipment. However, no studies have reported on the twinkling artifact of breast microcalcifications. While considering these premises, I detected microcalcifications in the breast using twinkling artifacts that could not be detected on B-mode imaging. The twinkling artifact is a well-defined but not well-understood phenomenon that may assist in the detection of calcified foci. The phenomenon of the twinkling artifact is discussed here with regard to prospectively and retrospectively studied cases including experiments with a new ultrasound modality called multidetector-ultrasonography (MD-US). MD-US using detectability of the twinkling artifact in microcalcifications of the breast may play an important role in breast screening.


Commentary on the effect of steroid use in anterior cervical discectomy and fusion surgery; a randomized controlled trial by Shiveindra B. et al. Journal of Neurosurgery Spine 2015;23:137-43.
Steroids are often used in patients undergoing anterior cervical discectomy and fusion (ACDF) surgery to limit postoperative dysphagia. However, a major concern remains steroids' impact is on fusion. In this prospective, randomized, double-blinded controlled study, the authors assessed the impact of steroids on swallowing/airway and fusion rates in 112 patients undergoing multilevel ACDF. The patients were randomly assigned to saline or dexamethasone groups prior to surgery; multiple other variables including different outcome analyses were also utilized over a 2-year postoperative period. The patients were followed for 1, 3, 6, 12, and 24 months postoperatively, and computed tomography (CT) studies were performed at 6, 12, and 24 postoperative months to establish fusion. The authors found no significant 2-year differences in the clinical parameters or surgical outcomes for patients undergoing ACDF with or without steroids. Steroids reduced dysphagia in the 1(st) postoperative month, produced a "trend" for reducing postoperative airway complications (e.g., intubation), and length of stay. Notably, CT-fusion rates with steroids were reduced at the 6(th) postoperative month but equalized by the 1(st) postoperative year. The authors concluded that dexamethasone administered at the time of ACDF surgery improved swallowing within the 1(st) postoperative month, reduced perioperative airway complications, reduced the length of stay, and reduced 6 month but not 12 month fusion rates. Although the findings regarding postoperative dysphagia are helpful, the performance of multiple 3D-CT scans postoperatively to document fusion would appear to subject these patients to excessive radiation exposure without sufficient clinical indications.


Isolated bilateral fibular insufficiency fractures: Case report of an unusual entity.
Insufficiency fractures are a type of stress fractures that occur due to physiological stress on weakened bones. These have certain sites of predilection of which an uncommon site is fibular shaft. Isolated bilateral fibular fractures have previously only been reported twice, where the causes were repeated trauma and fatigue fracture. Insufficiency fractures of both fibula in isolation have not previously been reported. We present this case where a young patient presented with these fractures and was found to have vitamin D deficiency on subsequent investigations.


The impact of leucine infusion on skeletal muscle amino acid and energy metabolism in severely traumatized patients.
The response to trauma is associated with increased energy requirements and net protein breakdown. The branched chain aminoacids, especially leucine, are considered to act by serving as a fuel for muscle tissue and by stimulating synthesis of proteins and controlling protein breakdown. Such results have been obtained mainly from in vitro studies. The present study was designed to evaluate the pharmacological effect of leucine infusion on muscle energy/amino acid metabolism in man after severe multiple trauma. 16 patients were studied and randomly allocated into 2 groups. Group 1 was given fat and 20% glucose while group 2 received 6 g N in form of leucine dissolved in 10% glucose solution and fat. The patients received 40 kcal/kg/24 h over an 8 day period after trauma. Biochemical analyses, muscle biopsies (energy substrates, electrolytes, amino acids), nitrogen balance and 3-methyl histidine excretion in urine were evaluated. Biochemical data revealed a significant increase (p < 0.05) of serum urea in group 2 day 4 and 8 after trauma. Muscle intracellular electrolytes (K(+), Mg(2+)) and energy substrates (ATP, phosphocreatine) showed a similar decrease in both groups. The intracellular muscle amino acids displayed a pattern known to be related to trauma without differences between the groups. The cumulative nitrogen balance 8 days after the injury was -93.5 g N +/- 10.1 (SEM) in group 1 and -73 g N +/- 7.5 in group 2. The 3-methylhistidine excretion was markedly increased similar in both groups. The present study demonstrated no significant pharmacological effect of leucine administration on muscle metabolism, nitrogen balance or 3-methylhistidine excretion in severely traumatized patients. Conventional balanced amino acid solutions are probably optimal to meet the patients actual requirements.


Study of the human male meiosis. I. G-banding in pachytene bivalents.
Results obtained from a meiotic study of 250 pachytene cells from four normal human males are presented. G-banding patterns for pachytene bivalents, obtained using trypsin treatment, are also presented; a comparative study between G-banding patterns of pachytene bivalents and G-banding patterns of mitotic chromosomes, and a study of chromomere counting and distribution, have also been made.


Molecular and biochemical investigation of the protective effects of rutin against liver and kidney toxicity caused by malathion administration in a rat model.
Widely used malathion (MLT) causes environmental pollution, leading to toxicity in many living things, including humans. Rutin (RUT) is a flavonoid with various biological properties. In the present study, the protective effects of rutin against liver and kidney toxicity caused by malathion were investigated. In the study, MLT (100 mg/kg) and RUT (50 or 100 mg/kg) were administered to rats alone or in combination for 28 days. Then, oxidative stress, inflammation, endoplasmic reticulum stress (ERS), apoptosis, and autophagy markers in liver and kidney tissues were analyzed by biochemical and molecular methods. The results showed that MLT caused oxidative stress in both tissues, while RUT showed antioxidant properties and protected these tissues from oxidative damage. Moreover, MLT upregulated the expressions of ATF-6, PERK, IRE1, GRP78, and CHOP, leading to ERS. However, RUT alleviated ER stress and suppressed these markers. The study also found that MLT increased inflammatory, apoptotic, and autophagic markers. All these factors affected liver and kidney functions and caused an increase in plasma ALT, AST, urea, and creatinine levels. On the other hand, it has been observed that RUT may protect liver and kidney tissues from the destructive effect of MLT by showing anti-inflammatory, anti-apoptotic, and anti-autophagic properties. Thus, it was determined that ALT, AST, urea, and creatinine levels decreased after RUT treatment. As a result, it was observed that MLT had a toxic effect on the liver and kidney tissues of rats, and it was determined that this toxicity could be alleviated by RUT treatment.


Randomized, double-masked, placebo-controlled comparison of the efficacy of emedastine difumarate 0.05% ophthalmic solution and ketotifen fumarate 0.025% ophthalmic solution in the human conjunctival allergen challenge model.
Emedastine difumarate 0.05% ophthalmic solution and ketotifen fumarate 0.025% ophthalmic solution are 2 topical antiallergic agents available in the United States and other countries. Emedastine is indicated for the temporary relief of the signs and symptoms of allergic conjunctivitis. Ketotifen is indicated for the temporary relief of ocular itching caused by allergic conjunctivitis. The purpose of this study was to compare the efficacy of these agents in the temporary relief of ocular itching due to allergic conjunctivitis. The 2 agents were compared with each other and with placebo (artificial tears) using the conjunctival allergen challenge (CAC) model. This was a single-center, randomized, double-masked, placebo-controlled study. At visit 1, CAC was performed on eligible subjects to identify the dose required to elicit a positive allergic reaction. Subjects returned after 7 days for visit 2 to confirm the allergen dose. On day 14 (+/-3) of the study, enrolled subjects were randomized to 1 of 3 treatment groups: emedastine in I eye and placebo in the other, ketotifen in 1 eye and placebo in the other, or emedastine in 1 eye and ketotifen in the other. In 25 subjects, bilateral CAC was performed 5 minutes after study medication instillation. In a second group of 20 subjects, CAC was performed 15 minutes after medication instillation. Itching was graded according to a standardized 5-point scale (0 = none to 4 = severe itching) at 3, 5, and 10 minutes postchallenge. Differences in efficacy scores between treatments and versus placebo were compared using 2-sample t tests of equal variance. A total of 45 patients (mean age, 41.2 years) received treatment: 16 received emedastine in 1 eye and ketotifen in the other; 14 received emedastine in 1 eye and placebo in the other; and 15 received ketotifen in 1 eye and placebo in the other. Both emedastine and ketotifen significantly inhibited itching (P < 0.05) compared with placebo at all time points after the 5- and 15-minute CAC. Mean raw scores for the active treatments were not statistically different. The mean itching efficacy scores were also not statistically different between active treatments. No adverse events were reported in this study. The results of this study suggest that emedastine and ketotifen are not significantly different with respect to anti-itching efficacy in the CAC model of acute allergic conjunctivitis.


Open Latarjet with Metal-Free Cerclage Fixation.
Despite multiple modifications, the Latarjet is still the most popular procedure for recurrent anterior shoulder instability with glenoid bone loss. Partial or subtotal resorption of the graft is common, potentially leading to hardware prominence and risk of anterior soft-tissue impingement. To minimize the technical difficulties and morbidity associated with metallic implants, a coracoid and conjoint tendon transfer with a mini-open approach using Cerclage tape suture is described, as an alternative for the Latarjet procedure typically performed with metal screws and plates.


Using banana to generate lactic acid through batch process fermentation.
We evaluated the usefulness of waste banana for generating lactic acid through batch fermentation, using Lactobacillus casei under three treatments. Two treatments consisted of substrates of diluted banana purée, one of which was enriched with salts and amino acids. The control treatment comprised a substrate suitable for L. casei growth. When fermentation was evaluated over time, significant differences (P<0.05) were found in the three treatments for each of five variables analyzed (generation and productivity of lactic acid, and consumption of glucose, fructose, and sucrose). Maximum productivity was (in g l(-1) h(-1)) 0.13 for the regular banana treatment, 1.49 for the enriched banana, and 1.48 for the control, with no significant differences found between the latter two treatments. Glucose consumption curves showed that L. casei made greater use of the substrate in the enriched banana and control treatments than in the regular banana treatment. For fructose intake, the enriched banana treatment showed significantly better (P<0.05) results than the regular one. Sucrose consumption was insignificant (P<0.05), probably because fermentation time was too short. Even when enriched, diluted banana purée is an ineffective substrate for L. casei, probably because it lacks nutrients.


Mitochondrial functions in plant immunity.
Mitochondria are energy factories of cells and are important for intracellular interactions with other organelles. Emerging evidence indicates that mitochondria play essential roles in the response to pathogen infection. During infection, pathogens deliver numerous enzymes and effectors into host cells, and some of these effectors target mitochondria, altering mitochondrial morphology, metabolism, and functions. To defend against pathogen attack, mitochondria are actively involved in changing intracellular metabolism, hormone-mediated signaling, and signal transduction, producing reactive oxygen species and reactive nitrogen species and triggering programmed cell death. Additionally, mitochondria coordinate with other organelles to integrate and amplify diverse immune signals. In this review, we summarize recent advances in understanding how mitochondria function in plant immunity and how pathogens target mitochondria for host defense suppression.


Prediction of dinucleotide-specific RNA-binding sites in proteins.
Regulation of gene expression, protein synthesis, replication and assembly of many viruses involve RNA-protein interactions. Although some successful computational tools have been reported to recognize RNA binding sites in proteins, the problem of specificity remains poorly investigated. After the nucleotide base composition, the dinucleotide is the smallest unit of RNA sequence information and many RNA-binding proteins simply bind to regions enriched in one dinucleotide. Interaction preferences of protein subsequences and dinucleotides can be inferred from protein-RNA complex structures, enabling a training-based prediction approach. We analyzed basic statistics of amino acid-dinucleotide contacts in protein-RNA complexes and found their pairing preferences could be identified. Using a standard approach to represent protein subsequences by their evolutionary profile, we trained neural networks to predict multiclass target vectors corresponding to 16 possible contacting dinucleotide subsequences. In the cross-validation experiments, the accuracies of the optimum network, measured as areas under the curve (AUC) of the receiver operating characteristic (ROC) graphs, were in the range of 65-80%. Dinucleotide-specific contact predictions have also been extended to the prediction of interacting protein and RNA fragment pairs, which shows the applicability of this method to predict targets of RNA-binding proteins. A web server predicting the 16-dimensional contact probability matrix directly from a user-defined protein sequence was implemented and made available at: http://tardis.nibio.go.jp/netasa/srcpred.


Promoter structure of human sonic hedgehog gene.
Sonic hedgehog (Shh) is a secreted signal transducer responsible not only for patterning of the anterior-posterior axis during early limb and neuronal development, but also for generating cell-type diversity at later developmental stages. To elucidate the mechanism regulating human Shh gene expression, we cloned the 5'-flanking region of the human Shh gene and characterized it by transient transfection studies. Two transcription start sites were identified by primer extension analysis. Two TATA-boxes, a CCAAT-box and a palindrome-like structure constituted the basic promoter structure. Furthermore, two continuous E-boxes and a putative homeodomain containing an ATTA-box were located around 350-450 bp upstream of the upper TATA-box. Consensus binding sites of the RA, estrogen, D3 and glucocorticoid/progesterone receptors were not found within the cloned sequence. Short-term treatment with TPA increased luciferase activity up to 2.1-fold; on the other hand, treatment with dibutyryl-cyclic AMP decreased it to 0.8-fold. Retinoic acid (RA), vitamin D3, dexamethazone (DEX) and estradiol (E2) had no effect on the luciferase activity. Since the zebrafish Shh promoter contains two closely spaced axial (HNF3beta) binding sequences on its basic promoter, the palindrome-like structure located in the corresponding site of the human Shh promoter may be a crucial binding domain regulating human Shh gene expression.


Effects of hypoxia-ischemia on monoamine metabolism in the immature brain.
We measured acute changes in monoamine metabolites in corpus striatum of immature rat pups exposed to hypoxia-ischemia, hypoxia alone, or total global ischemia. Carotid ligations and two hours of 8% oxygen environment in 7-day-old pups led to asymmetrical turning behavior, a 70% decrease in endogenous striatal dopamine levels, and a 125% increase in homovanillic acid (HVA) concentrations on the side of ligation. In contrast, hypoxia alone and total global ischemia alone were not associated with HVA level elevation. Elevation of HVA level with hypoxia-ischemia showed a threshold effect between 1 and 1.5 hours, and this time course paralleled that for production of gross morphological changes in rats raised to maturity. The data suggest that dopamine release from striatal nerve terminals is associated with events causing brain injury during perinatal hypoxia-ischemia. Tissue HVA in the animal model appears to be a quantitative marker for the effects of the insult on a population of nerve terminals.


Antiprogesterone and antiglucocorticoid actions of RU 486 on rabbit mammary gland explant cultures. Evidence for a persistent inhibitory action of residual progesterone upon the mammary tissue.
The antiprogesterone and antiglucocorticoid compound RU 486 added to pregnant rabbit mammary gland explant cultures had no effect alone but significantly stimulated casein production in the presence of ovine prolactin (PRL) in a dose dependent manner. This stimulation was inhibited by progesterone (Pg) and the Pg agonist R5020. When the explants were cultured for 5 days with two changes of medium, to eliminate all steroids, and hormones added afterwards, the effect of PRL was potentiated, Pg was no longer inhibitory and RU 486 had no effect, RU 486 also could inhibit the stimulatory action of glucocorticoids added to the cultures along with PRL. The compound was able to displace [3H]dexamethasone and [3H]R 5020 from mammary gland glucocorticoid and Pg receptors respectively and proved to have a high relative binding affinity (RBA) for both receptors when compared with typical ligands for each receptor. The RBAs of RU 486 and the steroids used in this study to mammary gland glucocorticoid and Pg receptors correlated well with the ability of RU 486 to block their biological activities. These results demonstrate that RU 486 has both antiglucocorticoid and antiprogesterone activities in pregnant rabbit mammary glands as well as the existence of a strong inhibitory residual action of Pg in the gland that persists during the first 48 h of culture and that can be eliminated by RU 486 or after several days of culture with no hormones.


Systemically administered adenosine increases caudate blood flow in rabbits.
Adenosine has been proposed to be a chemical link between cerebral metabolism and blood flow. In the present study, we investigated whether the intravenous or intracarotid administration of adenosine could influence regional cerebral blood flow in anesthetized rabbits. The study was performed with the [14C]ethanol tissue sampling technique which enables quantitative, instantaneous, multiregional blood flow measurements. With either mode of adenosine administration, no change in cerebral blood flow was observed, except in the caudate nucleus in which a significant vasodilation took place. These data indicate that, in rabbits exogenous adenosine increases blood flow in highly specific brain areas, by mechanisms that are discussed.


Identification, coassembly, and activity of gamma-aminobutyric acid receptor subunits in renal proximal tubular cells.
Although the properties and functions of GABA(A) receptors in the mammalian central nervous system have been well studied, the presence and significance of GABA(A) receptors in non-neural tissues are less clear. The goal of this study was to examine the expression of GABA(A) receptor alpha(1), alpha(2), alpha(4), alpha(5), beta(1), gamma(1), gamma(2), and delta subunits in the kidney and to determine whether these subunits coassemble to form an active renal epithelial cell GABA(A) receptor. Using reverse transcriptase products from RNA isolated from rat and rabbit kidney cortex and brain or cerebellum through polymerase chain reaction (PCR) and sequencing of the PCR products, we revealed that rat kidney cortex contained the alpha(1), alpha(5), beta(1), gamma(1), and gamma(2) subunits and that they were similar to the neuronal subunits. Sequencing of the PCR products revealed that the rabbit kidney cortex contained the alpha(1) and gamma(2) subunits and that they were similar to their neuronal counterparts. Immunoprecipitation and immunoblot studies using GABA(A) receptor subunit-specific antibodies and detergent-solubilized rat kidney cortex membranes identified a GABA(A) receptor complex containing alpha(5), beta(1), and gamma(1). Isolated rat renal proximal tubular cells exhibited GABA-mediated, picrotoxin-sensitive (36)Cl(-) uptake. These studies demonstrate the presence of numerous GABA(A) receptor subunits in the kidneys of two species, the assembly of the subunits into at least one novel receptor complex, and an active GABA(A) receptor in renal proximal tubular cells.


Histochemistry of the post-resuscitation changes in the brain.
In 20 dogs, acid mucopolysaccharides, gangliosides, lysosomal enzymes and iontransport enzymes were investigated. Material was taken 2, 24 and 72 h after 15 min of clinical death due to electrotrauma. Changes in the post-resuscitation period indicated deterioration of transmembrane ion--metabolite transport in the brain. These changes had phasic variations that reflect the complexity of post-resuscitation processes.


Single case study of brain FDG PET imaging in a patient with catatonia.
We report a case of bilateral occipitotemporal and thalamic hypometabolism on FDG PET in a 19-year-old male patient who presented with altered mental status. He had a history of static encephalopathy and presented with visual hallucinations and decreased verbal output, sleep, and appetite 7 days after starting amoxicillin for otitis media. Extensive evaluation failed to demonstrate any infectious, inflammatory, autoimmune-mediated (including negative testing for NMDA receptor antibody), paraneoplastic, or toxic-metabolic etiology. Ultimately, he responded well to a benzodiazepine challenge and electroconvulsive therapy and was diagnosed with catatonia.


Efficacy of a live intranasal vaccine against parainfluenza type 3 and bovine respiratory syncytial virus in young calves with maternally derived antibodies.
Two randomised controlled vaccination trials with artificial challenges were carried out in addition to a serological survey of levels of maternally derived antibodies (MDA) to parainfluenza type 3 virus (PI3V) and bovine respiratory syncytial virus (BRSV) in European calves. Ten-day-old calves with and without MDA were included in the two vaccine trials. Intranasal administration of a bivalent modified live (PI3V/BRSV) vaccine followed by artificial challenge approximately three months post vaccination. The study aimed to assess the efficacy of a modified live respiratory vaccine, Bovalto Respi Intranasal (Boehringer Ingelheim). In order to assess the interference of MDA, both seropositive and seronegative calves were used. PI3V and BRSV serological status was determined seven days before vaccination; calves without maternal antibodies became the MDA- vaccinates. Calves with MDA were ranked according to individual titres and allocated alternately to MDA+ vaccinate and MDA+ control groups. Treatment was carried out by the unblinded study director. Animal care and veterinary examinations were conducted by personnel unaware of the treatments received. The serological survey used blood samples obtained from calves on commercial farms in five European countries, Germany, Spain, Italy, Ireland and the UK, to determine the levels of MDA to PI3V and BRSV in calves approximately two weeks of age. A total of 36 calves were included in the two challenge studies and 32 of these completed the challenge studies. Twenty-one calves were included in the PI3V challenge study, with six of six MDA- and six of seven MDA+ vaccinated calves and five of five MDA+ unvaccinated control calves being challenged with PI3V. Fifteen calves were included in the BRSV challenge study, with five of five MDA- and five of five MDA+ vaccinated calves and five of five MDA+ unvaccinated control calves being challenged with BRSV. For both challenges, clinical scores and nasal shedding were significantly higher in control animals compared with vaccinates (PI3V challenge: clinical scores P=0.001, nasal shedding P=0.001; BRSV challenge: clinical scores P=0.016, nasal shedding P=0.002) and not significantly different between MDA+ and MDA- vaccinated animals for both challenges (P>0.05). A total of 254 samples from six countries were tested in the serological survey of MDA. The results of the challenge studies demonstrated the efficacy of the vaccine in the presence of BRSV and PI3V MDA under laboratory conditions. The field assessment confirmed that the MDA titres in the MDA+ calves corresponded to those typically found on farms.


Characteristics of advanced porous and textured surface pacemaker electrodes.
Reduced stimulation thresholds, improved sensing and better attachment have been claimed for totally porous and porous surfaced electrodes. In this study, the potential for clinical use of two new types of porous electrodes and a non-porous, textured high microsurface area electrode, has been evaluated by comparison with equivalent sized, smooth non-porous controls. Eighteen sintered and seven laser drilled porous electrodes, seven-non-porous textured electrodes, and sixteen controls, were implanted singly in the right ventricles of sheep. Measurements of threshold, pacing, sensing and bulk impedances were taken at regular intervals for up 180 days. At sacrifice, only three of the thirteen non-porous controls were attached. All laser porous electrodes, apart from two which were dislodged, were attached, as were eleven of fifteen sintered porous, and five of seven textured non-porous electrodes. Tissue ingrowth was found for both porous electrode types. Stimulation thresholds were not statistically different for all electrode types (p less than 0.05). Pacing and bulk impedances of the two porous and surface textured electrodes were significantly higher (p less than 0.10, p less than 0.05, respectively) than those of controls. The three new electrodes exhibited similar chronic sensing impedance values, 30% less than equivalent non-porous electrodes. The similar sensing performance of the porous and high microsurface area non-porous electrodes indicates that the nature of the external surface, rather than internal porosity, determines sensing impedance. All three new electrode types showed improved attachment and sensing compared with similar smooth electrodes. The laser porous electrode, which permits fixation by tissue ingrowth and maintains simplicity of construction, is promising for routine clinical use.


[Magnetic resonance tomography imaging of Botallo's duct--a case report].
In a female patient suffering from renal insufficiency in whom angiography could not be performed, it was possible to confirm the diagnosis of a ductus arteriosus (Botallo's duct) by magnetic resonance imaging. The present case report shows that it is possible to visualise this congenital heart disease by MRI. This can be of importance especially if angiography cannot be performed (as in this case) because of renal insufficiency or because of contrast medium allergy.


Label-Free Visualization and Quantification of Biochemical Markers of Atherosclerotic Plaque Progression Using Intravascular Fluorescence Lifetime.
This study aimed to systematically investigate whether plaque autofluorescence properties assessed with intravascular fluorescence lifetime imaging (FLIm) can provide qualitative and quantitative information about intimal composition and improve the characterization of atherosclerosis lesions. Despite advances in cardiovascular diagnostics, the analytic tools and imaging technologies currently available have limited capabilities for evaluating in situ biochemical changes associated with luminal surface features. Earlier studies of small number of samples have shown differences among the autofluorescence lifetime signature of well-defined lesions, but a systematic pixel-level evaluation of fluorescence signatures associated with various histological features is lacking and needed to better understand the origins of fluorescence contrast. Human coronary artery segments (n = 32) were analyzed with a bimodal catheter system combining multispectral FLIm with intravascular ultrasonography compatible with in vivo coronary imaging. Various histological components present along the luminal surface (200-μm depth) were systematically tabulated (12 sectors) from each serial histological section (n = 204). Morphological information provided by ultrasonography allowed for the accurate registration of imaging data with histology data. The relationships between histological findings and FLIm parameters obtained from 3 spectral channels at each measurement location (n = 33,980) were characterized. Our findings indicate that fluorescence lifetime from different spectral bands can be used to quantitatively predict the superficial presence of macrophage foam cells (mFCs) (area under the receiver-operator characteristic curve: 0.94) and extracellular lipid content in advanced lesions (lifetime increase in 540-nm band), detect superficial calcium (lifetime decrease in 450-nm band area under the receiver-operator characteristic curve: 0.90), and possibly detect lesions consistent with active plaque formation such as pathological intimal thickening and healed thrombus regions (lifetime increase in 390-nm band). Our findings indicate that autofluorescence lifetime provides valuable information for characterizing atherosclerotic lesions in coronary arteries. Specifically, FLIm can be used to identify key phenomena linked with plaque progression (e.g., peroxidized-lipid-rich mFC accumulation and recent plaque formation).


Detection of Kaposi sarcoma associated herpesvirus in peripheral blood of HIV-infected individuals and progression to Kaposi's sarcoma.
Kaposi sarcoma-associated herpesvirus (KSHV) is consistently found in biopsy samples from patients with AIDS-related and "classical" Kaposi's sarcoma (KS). Although highly suggestive of a causal role of KSHV in the pathogenesis of KS, this observation does not exclude the possibility that KSHV, like other herpesviruses, is widely distributed and is a mere "passenger" in these lesions. Here we report that KSHV was detectable in peripheral blood mononuclear cells of 24/46 (52%) of KS patients, but in none of 134 blood donors or 26 HIV-uninfected hospital controls. KSHV detection increased with immunosuppression, as shown by a correlation with a reduced number of CD4-positive T-cells. Moreover, KSHV detection in peripheral blood cells of HIV-infected individuals without KS predicted the subsequent appearance of KS lesions. 143 patients who did not have KS at the time of their first (or only) blood sample were followed up for a median of 30 months. Of the 11 who had been KSHV positive 6 developed KS compared with only 12 out of 132 who were KSHV negative. These findings are compatible with a causative role of KSHV in KS. KSHV was rarely detected in sputum and throat swabs of HIV-infected patients, providing a potential explanation for the apparently limited spread of this virus.


Progestin receptors from tissues either exhibiting or lacking estrogen response mechanisms. Comparison of conventional and high-performance liquid chromatography methodology.
Evidence from a variety of target organs has shown that progesterone receptor (PR) is induced by estrogen receptor (ER) in normal and neoplastic tissues. However, approximately 12% of the normal human uterine samples exhibit only PR with no measurable ER, suggesting the expression of both inducible and constitutive receptor isoforms. We investigated several molecular properties of PR from tissues either exhibiting or lacking ER. All studies were conducted in potassium phosphate buffer containing 10 mM sodium molybdate with a synthetic progestin, [3H]R5020 as the ligand. Radioinert R5020 was used as competitor to assess nonspecific association. Competition analysis showed that PR from both sources exhibited similar ligand specificities and affinities. Relative affinities were ORG 2058 greater than R5020 greater than medroxy-progesterone acetate greater than progesterone much greater than testosterone (Kd values ranged from 10(-9) to 10(-10) M; testosterone showed no specific competition). We utilized high-performance liquid chromatography in the size-exclusion (HPSEC) and ion-exchange (HPIEC) modes to probe the size and ionic properties of PR. HPSEC profiles showed that the PR isoform from both sources was eluted as a single, sharp peak greater than 75 A. HPIEC elution profiles indicated no differences in the surface ionic properties in that PR from both tissue types eluted with ca. 100 mM phosphate. These experiments show no difference between the inducible and the putative constitutive form of PR. Thus, some PR species may not require estrogen for their formation.


Ecosystem ecology: size-based constraints on the pyramids of life.
Biomass distribution and energy flow in ecosystems are traditionally described with trophic pyramids, and increasingly with size spectra, particularly in aquatic ecosystems. Here, we show that these methods are equivalent and interchangeable representations of the same information. Although pyramids are visually intuitive, explicitly linking them to size spectra connects pyramids to metabolic and size-based theory, and illuminates size-based constraints on pyramid shape. We show that bottom-heavy pyramids should predominate in the real world, whereas top-heavy pyramids indicate overestimation of predator abundance or energy subsidies. Making the link to ecological pyramids establishes size spectra as a central concept in ecosystem ecology, and provides a powerful framework both for understanding baseline expectations of community structure and for evaluating future scenarios under climate change and exploitation.


The VP3 protein of duck hepatitis A virus mediates host cell adsorption and apoptosis.
Duck hepatitis A virus (DHAV) causes an infectious disease that mainly affects 1- to 4-week-old ducklings, resulting in considerable loss to the duck industry. Although there have been many studies on DHAV in recent years, the effects on host infection and pathogenesis of DHAV-1 remain largely unknown. This study investigated the effects of the DHAV-1 structural protein VP3 on DHAV-1 virus adsorption and apoptosis to explore the role of VP3 in the viral life cycle. The effects of DHAV-1 VP3 and an antibody against the protein on virion adsorption was analyzed by qRT-PCR. The results showed that the virus copy number for the rabbit anti-VP3 IgG-treated group was significantly lower than that for the negative control group but higher than that for the rabbit anti-DHAV-1 IgG-treated group. This result indicates that VP3 mediates DHAV-1 virus adsorption but that it is not the only protein that involved in this process. In addition, a eukaryotic recombinant plasmid, pCAGGS/VP3, was transfected into duck embryo fibroblasts (DEFs), and the apoptotic rate was determined by DAPI staining, the TUNEL assay and flow cytometry. DAPI staining showed nucleus fragmentation and nuclear edge shifting. TUNEL assay results revealed yellow nuclei, and flow cytometry indicated a significant increase in the apoptotic rate. In addition, qRT-PCR revealed increased in the transcriptional levels of the apoptotic caspase-3, -8 and -9, with the largest increase for caspase-3, followed by caspase-9 and caspase-8. Enzyme activity analysis confirmed these results. Furthermore, the VP3 protein decreased the mitochondrial membrane potential, and the transcriptional levels of the proapoptotic factors Bak, Cyt c and Apaf-1 in the mitochondrial apoptotic pathway were significantly upregulated. These data suggest that expression of VP3 in DEFs induces apoptosis and may primarily activate caspase-3-induced apoptosis through mitochondrion-mediated intrinsic pathways. The findings provide scientific data to clarify DHAV-1 infection and pathogenesis.


Polyradiculoneuritis and Mycoplasma pneumoniae infection.
A patient with severe Mycoplasma pneumonia developed polyradiculoneuritis and respiratory failure. The acute phase of the illness was complicated by a myocarditis, and recovery of neurological function was slow. Residual left hemidiaphragmatic paralysis was present 1 year after onset of the illness.


Taxanes in adjuvant breast cancer setting: which standard in Europe?
The clinical studies of cooperators groups (trials CALGB 9344, NSABP-B-28, BCIRG 001, PACS01 and CALGB 9741) demonstrated, in the adjuvant breast cancer setting, that taxanes (paclitaxel and docetaxel) improved both disease free and overall survival (trials CALGB 9344, BCIRG 001 and PACS 01). However, the debate remains open in Europe. Less than 50% of the expert present at the last St. Gallen Conference recommended the use of taxanes in adjuvant setting. The reasons for this are primarily related to the fact that the comparator arms of cooperators group (AC, FAC and FEC 100) are considered by some Europe groups as being less effective than the European standards (chemotherapy (CMF) and Epirubicine-CMF). Many questions remain unanswered, including whether the use of taxanes should be sequential or concomitant, and which population would benefit from such a treatment: patients with hormone-receptor negative disease and/or the HER-2 positive tumors?


Quantification of Carotenoid Pathway Flux in Green and Nongreen Systems.
Metabolite accumulation in plant tissues represents the transient net result of their constant biosynthesis and degradation. For carotenoids, degradation might occur enzymatically by carotenoid cleavage producing plant hormones and volatiles or by nonenzymatic oxidation, both depending on environmental and developmental conditions. Carotenoid biosynthesis is therefore constantly regulated at various levels to attain sufficient carotenoid accumulation, mainly for photosynthesis and photoprotection. Due to the plenitude of carotenoids and their degradation products, it is not feasible to investigate overall carotenoid biosynthetic activity and its regulation by the quantification of all carotenoids including their derivatives. This is an issue encountered in investigations on many other highly branched pathways. We therefore present protocols to determine carotenoid biosynthesis flux in a given plant tissue by HPLC quantification of phytoene, the first pathway-specific intermediate and precursor of all carotenoids synthesized by phytoene synthase (PSY). For this purpose, enzymatic metabolization of phytoene in the tissue under investigation is prevented by treatment with the bleaching herbicide norflurazon, blocking the carotenogenic pathway downstream of PSY. As phytoene is more resistant to oxidation than desaturated carotenoids, the rate of phytoene biosynthesis serves as a good measure for total carotenogenic flux in a given tissue. The method is described for Arabidopsis for two photosynthetically active sample types, namely, seedlings and leaves, as well as for seed-derived callus as nongreen tissue. It should be realizable using only a relatively simple experimental setup and is applicable to other plant tissues as well as to different plant species. Additionally, similar experimental setups could be a useful tool to investigate total flux and turnover rates in other biosynthetic pathways.


Truncated thioredoxin (Trx-80) promotes pro-inflammatory macrophages of the M1 phenotype and enhances atherosclerosis.
Vascular cells are particularly susceptible to oxidative stress that is believed to play a key role in the pathogenesis of cardiovascular disorders. Thioredoxin-1 (Trx-1) is an oxidative stress-limiting protein with anti-inflammatory and anti-apoptotic properties. In contrast, its truncated form (Trx-80) exerts pro-inflammatory effects. Here we analyzed whether Trx-80 might exert atherogenic effects by promoting macrophage differentiation into the M1 pro-inflammatory phenotype. Trx-80 at 1 µg/ml significantly attenuated the polarization of anti-inflammatory M2 macrophages induced by exposure to either IL-4 at 15 ng/ml or IL-4/IL-13 (10 ng/ml each) in vitro, as evidenced by the expression of the characteristic markers, CD206 and IL-10. By contrast, in LPS-challenged macrophages, Trx-80 significantly potentiated the differentiation into inflammatory M1 macrophages as indicated by the expression of the M1 cytokines, TNF-α and MCP-1. When Trx-80 was administered to hyperlipoproteinemic ApoE2.Ki mice at 30 µg/g body weight (b.w.) challenged either with LPS at 30 µg/30 g (b.w.) or IL-4 at 500 ng/30 g (b.w.), it significantly induced the M1 phenotype but inhibited differentiation of M2 macrophages in thymus and liver. When ApoE2.Ki mice were challenged once weekly with LPS for 5 weeks, they showed severe atherosclerotic lesions enriched with macrophages expressing predominantly M1 over M2 markers. Such effect was potentiated when mice received daily, in addition to LPS, the Trx-80. Moreover, the Trx-80 treatment led to a significantly increased aortic lesion area. The ability of Trx-80 to promote differentiation of macrophages into the classical proinflammatory phenotype may explain its atherogenic effects in cardiovascular diseases.


Pathways to cellular supremacy in biocomputing.
Synthetic biology uses living cells as the substrate for performing human-defined computations. Many current implementations of cellular computing are based on the "genetic circuit" metaphor, an approximation of the operation of silicon-based computers. Although this conceptual mapping has been relatively successful, we argue that it fundamentally limits the types of computation that may be engineered inside the cell, and fails to exploit the rich and diverse functionality available in natural living systems. We propose the notion of "cellular supremacy" to focus attention on domains in which biocomputing might offer superior performance over traditional computers. We consider potential pathways toward cellular supremacy, and suggest application areas in which it may be found.


Accelerating effect of pyocyanin on microbiologically influenced corrosion of 304 stainless steel by the Pseudomonas aeruginosa biofilm.
In this study, the influence of exogenous pyocyanin (PYO) on the microbiologically influenced corrosion (MIC) of 304 stainless steel by Pseudomonas aeruginosa was investigated. Under sterile condition, the additional PYO in the culture medium had no effect on the corrosion of 304 stainless steel. In contrast, P. aeruginosa biofilm inoculated in the media with additional PYO resulted in more severe pitting corrosion. EIS and cyclic potentiodynamic polarization results indicated that exogenous PYO promoted the electron transfer efficiency between the P. aeruginosa biofilm and the stainless steel surface. X-ray photoelectron spectroscopy (XPS) and transmission electron microscope (TEM) results further demonstrated that the P. aeruginosa led the breakdown of passive film predominantly by accelerating the bioreductive dissolution of iron oxides.


Preoperative autologous blood donation in primary total knee arthroplasty: critical review of current indications.
Preoperative autologous blood donation (PABD) is a well established transfusion practice in elective orthopaedic surgery, involving immunologic and infective advantages but also involving exposure to not negligible risks, and costs as well. The aim of this study was to assess the real need for blood transfusions in primary total knee arthroplasty (TKA). Between January 2000 and July 2005, 214 patients underwent primary unilateral TKA. Altogether, 416 autologous blood units were collected, however only 47 (11.3%) were transfused. Thirty-eight patients (17.8%) received autologous blood, while 4 of them (10.5%) also received allogeneic blood. Based on the results of this study, PABD should be recommended in well selected patients undergoing TKA: older female patients with a low basal haemoglobin level.


Nuclear cytomorphometry in feline mammary gland epithelial tumours.
Stained cytological specimens from 35 feline mammary gland epithelial tumours (4 adenomas, 11 tubulopapillary carcinomas 13 solid carcinomas and 7 cribriform carcinomas) were analysed by computer-assisted nuclear morphometry. In each case, the nuclei of at least 100 neoplastic cells were measured, and the mean nuclear area (MNA), mean nuclear perimeter (MNP), mean nuclear diameter (MND) and nuclear roundness (NR) were calculated. The study aimed to evaluate (1) the possibility of using nuclear cytomorphometry as an auxiliary diagnostic method to differentiate between benign and malignant feline mammary gland epithelial tumours, and (2) the prognostic value of nuclear morphometry in feline mammary carcinomas. The results indicated that MNA, MNP, MND and NR could be a useful adjunct in diagnosis but are not reliable prognostic indicators for feline mammary gland carcinomas.


Transcriptomic profiles of the smoke tree wilt fungus Verticillium dahliae under nutrient starvation stresses.
Verticillium dahliae is a notorious plant pathogen that causes vascular wilt on more than 200 plant species. During plant infection, efficient pathogen nutrition during the interaction with the host is a requisite for successful infection. However, little attention has been focused on nutrient uptake and starvation responses in this fungus. Here, we used RNA-Seq to analyze the response of V. dahliae to nutrient starvation, including carbon and nitrogen depletion. Gene expression profile analysis showed that 1854 genes were differentially expressed under carbon starvation (852 upregulated and 539 downregulated genes) and nitrogen starvation (487 upregulated and 291 downregulated genes). Among the differentially expressed genes, genes involved in utilization or production acetyl-CoA, including glycolysis, fatty acid biosynthesis or metabolism, and melanin biosynthesis, were repressed under carbon starvation, whereas melanin biosynthesis genes were strongly induced under nitrogen starvation. These results, combined with VDH1 expression data, suggested that melanin biosynthesis and microsclerotia development were induced under nitrogen starvation, but microsclerotia development was suppressed under carbon starvation. Furthermore, many genes encoding carbohydrate-active enzymes and secreted proteins were induced under carbon starvation. Overall, the results improve our understanding of the response of V. dahliae to nutrient starvation and help to identify potential virulence factors for the development of novel disease control strategies.


Intrinsic bioactivity of thyrotropin in human serum is inversely correlated with thyroid hormone concentrations. Application of a new bioassay using the FRTL-5 rat thyroid cell strain.
We have developed a new bioassay for thyrotropin (TSH) in human serum to evaluate bioactivity in normal individuals and patients with different degrees of primary hypothyroidism. Unpurified TSH in serum showed no stimulation of cyclic AMP production in cultured FRTL-5 rat thyroid cells, but after immunopurification showed potent stimulatory activity. Immunoaffinity purification permitted up to 400-fold concentration of serum TSH, allowing bioactivity measurements even in certain normal sera. The limit of detection in the FRTL-5 bioassay was 10 microU of human TSH per 0.5 ml incubate, and half-maximal responses for standard human TSH was 102 +/- 26 (+/- SE) microU/0.5 ml. Immunoaffinity-purified serum TSH varied in bioactivity-to-immunoactivity (B/I) ratios from less than 0.25 to 1.21 among four euthyroid subjects and eight primary hypothyroid patients. An inverse correlation was found between B/I ratios of immunopurified basal TSH and the serum-free T4 (r = -0.7237, P less than 0.01), T4 (r = -0.6650, P less than 0.05), and T3 (r = -0.6382, P less than 0.05). B/I ratios of immunopurified TSH from three hypothyroid patients before and after acute stimulation by thyrotropin-releasing hormone showed no significant change, despite major changes in serum TSH. In summary, the present study shows an inverse relationship between the metabolic status of an individual and the intrinsic bioactivity of TSH.


Enzymatic activities of muscle fibres differentiated, in vitro, from pectoralis major (white) and adductor magnus (red) muscles of chick embryos.
Specific activities of NADP isocitrate dehydrogenase and acetylcholinesterase were significantly higher in muscle fibres differentiated, in vitro, from myoblasts of adductor magnus (red) than pectoralis major (white) muscles 10-day-old chick embryos. This is evidence, as far as enzyme activities are concerned, that myoblasts from different types of skeletal muscles are able to give, in tissue culture, muscle fibres of different properties, even in the absence of nerve supply.