Generation of Tn5 insertions in streptococcal conjugative transposon Tn916.
A method has been developed for the introduction of Tn5 into Escherichia coli plasmid chimeras containing Streptococcus faecalis DNA. Tn5 could be introduced via a lambda::Tn5 delivery vehicle. The system proved to be particularly efficient and facilitated insertions at numerous sites on DNA containing the 16-kilobase conjugative transposon Tn916. It was possible to introduce some of the resulting Tn916::Tn5 derivatives back into S. faecalis by using a recently developed protoplast transformation procedure. A presumed zygotic induction resulted in insertion of the Tn916 derivatives at multiple sites in the S. faecalis chromosome.


Modelling the role of immunity in reversion of viral antigenic sites.
Antigenic sites in viral pathogens exhibit distinctive evolutionary dynamics due to their role in evading recognition by host immunity. Antigenic selection is known to drive higher rates of non-synonymous substitution; less well understood is why differences are observed between viruses in their propensity to mutate to a novel or previously encountered amino acid. Here, we present a model to explain patterns of antigenic reversion and forward substitution in terms of the epidemiological and molecular processes of the viral population. We develop an analytical three-strain model and extend the analysis to a multi-site model to predict characteristics of observed sequence samples. Our model provides insight into how the balance between selection to escape immunity and to maintain viability is affected by the rate of mutational input. We also show that while low probabilities of reversion may be due to either a low cost of immune escape or slowly decaying host immunity, these two scenarios can be differentiated by the frequency patterns at antigenic sites. Comparison between frequency patterns of human influenza A (H3N2) and human RSV-A suggests that the increased rates of antigenic reversion in RSV-A is due to faster decaying immunity and not higher costs of escape.


Glycolysis Inhibitors for Anticancer Therapy: A Review of Recent Patents.
The aerobic glycolysis in tumor cells known as Warburg effect is one of the most important hallmarks of cancer. It is proposed that the upregulation of the series of metabolic enzymes along the glycolytic pathway may contribute to the Warburg effect. The inhibition of these glycolytic enzymes has been found to be a novel strategy for anticancer treatment. This review summaries recent patents in the development of small molecule inhibitors for the key enzymes in tumor glycolysis. The targeted enzymes are GLUTs, HKs, PFK, PGAM1, PKM2, LDHA, MCTs and PDK. Although most inhibitors are still in the preclinical phase, the inhibition of glycolytic enzymes represents a very promising approach for anticancer treatment. The future development could be more focused on the discovery of new metabolic enzyme that is specifically expressed in tumor cells.


Regulation of alpha(2)-adrenoceptors in human vascular smooth muscle cells.
This study analyzed the regulation of alpha2-adrenoceptors (alpha2-ARs) in human vascular smooth muscle cells (VSMs). Saphenous veins and dermal arterioles or VSMs cultured from them expressed high levels of alpha2-ARs (alpha2C > alpha2A, via RNase protection assay) and responded to alpha2-AR stimulation [5-bromo-N-(4,5-dihydro-1H-imidazol-2-yl)-6-quinoxalinamine (UK-14,304, 1 microM)] with constriction or calcium mobilization. In contrast, VSMs cultured from aorta did not express alpha2-ARs and neither cultured cells nor intact aorta responded to UK-14,304. Although alpha2-ARs (alpha2C >> alpha2A) were detected in aortas, alpha2C-ARs were localized by immunohistochemistry to VSMs of adventitial arterioles and not aortic media. In contrast with aortas, aortic arterioles constricted in response to alpha2-AR stimulation. Reporter constructs demonstrated higher activities for alpha2A- and alpha2C-AR gene promoters in arteriolar compared with aortic VSMs. In arteriolar VSMs, serum increased expression of alpha2C-AR mRNA and protein but decreased expression of alpha2A-ARs. Serum induction of alpha2C-ARs was reduced by inhibition of p38 mitogen-activated protein kinase (MAPK) with 2 microM SB-202190 or dominant-negative p38 MAPK. UK-14,304 (1 microM) caused calcium mobilization in control and serum-stimulated cells: in control VSMs, the response was inhibited by the alpha2A-AR antagonist BRL-44408 (100 nM) but not by the alpha2C-AR antagonist MK-912 (1 nM), whereas after serum stimulation, MK-912 (1 nM) but not BRL-44408 (100 nM) inhibited the response. These results demonstrate site-specific expression of alpha2-ARs in human VSMs that reflects differential activity of alpha2-AR gene promoters; namely, high expression and function in venous and arteriolar VSMs but no detectable expression or function in aortic VSMs. We found that alpha2C-ARs can be dramatically and selectively induced via a p38 MAPK-dependent pathway. Therefore, altered expression of alpha2C-ARs may contribute to pathological changes in vascular function.


Breast cancer incidence, 1980-2006: combined roles of menopausal hormone therapy, screening mammography, and estrogen receptor status.
Breast cancer incidence has been rising since at least 1935-1939, but recent US data reveal a statistically significant decline in breast cancer incidence in 2003 that persisted through 2004. Identifying the specific contributions of the potential causes of this long-term increase and the recent decrease in incidence has been challenging. Marked changes in rates of mammography screening and use of menopausal hormone therapy since 1980 have added further complexity. We examined the potential association between menopausal hormone therapy use and recent changes in breast cancer incidence. Using tumor registry, clinical, pathology, and pharmacy data from Kaiser Permanente Northwest, a large prepaid US health plan, we compared age-specific and age-adjusted breast cancer incidence rates (2-year moving averages) with use of screening mammography and dispensed menopausal hormone therapy prescriptions between 1980 and 2006. Temporal changes in incidence rates were assessed via joinpoint regression. A total of 7386 incident invasive breast cancers were diagnosed in plan members from 1980 through 2006. Overall age-adjusted breast cancer incidence rates per 100,000 women rose 25% from the early 1980s (105.6) to 1992-1993 (131.7) and an additional 15% through 2000-2001 (151.3), then dropped by 18% to 2003-2004 (123.6) and edged up slightly in 2005-2006 (126.2). These patterns were largely restricted to women aged 45 years or older and to estrogen receptor-positive (ER+) breast cancers. Incidence rates of ER-negative tumors experienced neither of the rises seen for ER+ tumors but also fell precipitously from 2003 through 2006. Rates of mammography screening sharply increased from 1980 to 1993 but then leveled off, and 75%-79% of women aged 45 years or older received a mammogram at least once every 2 years from 1993 through 2006. Menopausal hormone therapy dispensings, particularly of estrogen-plus-progestin formulations, increased from 1988 to 2002 but then dropped by approximately 75% after 2002. From 1980 through 2006, quantitative and qualitative trends in breast cancer incidence rates, particularly for ER+ tumors, parallel major changes in patterns of mammography screening and use of menopausal hormone therapy.


Impact of insufficient sleep on total daily energy expenditure, food intake, and weight gain.
Insufficient sleep is associated with obesity, yet little is known about how repeated nights of insufficient sleep influence energy expenditure and balance. We studied 16 adults in a 14- to 15-d-long inpatient study and quantified effects of 5 d of insufficient sleep, equivalent to a work week, on energy expenditure and energy intake compared with adequate sleep. We found that insufficient sleep increased total daily energy expenditure by ∼5%; however, energy intake--especially at night after dinner--was in excess of energy needed to maintain energy balance. Insufficient sleep led to 0.82 ± 0.47 kg (±SD) weight gain despite changes in hunger and satiety hormones ghrelin and leptin, and peptide YY, which signaled excess energy stores. Insufficient sleep delayed circadian melatonin phase and also led to an earlier circadian phase of wake time. Sex differences showed women, not men, maintained weight during adequate sleep, whereas insufficient sleep reduced dietary restraint and led to weight gain in women. Our findings suggest that increased food intake during insufficient sleep is a physiological adaptation to provide energy needed to sustain additional wakefulness; yet when food is easily accessible, intake surpasses that needed. We also found that transitioning from an insufficient to adequate/recovery sleep schedule decreased energy intake, especially of fats and carbohydrates, and led to -0.03 ± 0.50 kg weight loss. These findings provide evidence that sleep plays a key role in energy metabolism. Importantly, they demonstrate physiological and behavioral mechanisms by which insufficient sleep may contribute to overweight and obesity.


Combination of a mood stabilizer with risperidone or haloperidol for treatment of acute mania: a double-blind, placebo-controlled comparison of efficacy and safety.
The study assessed the efficacy and safety of risperidone as an adjunctive agent to mood stabilizers in the treatment of acute mania. This 3-week randomized, double-blind, placebo-controlled study included 156 bipolar disorder patients with a current manic or mixed episode who received a mood stabilizer (lithium or divalproex) and placebo, risperidone, or haloperidol. The primary efficacy measure was the Young Mania Rating Scale. Other assessments used the Brief Psychiatric Rating Scale, the Clinical Global Impression scale, and safety measures. The trial was discontinued by 25 (49%) of the 51 placebo group patients, 18 (35%) of the 52 risperidone group patients, and 28 (53%) of the 53 haloperidol group patients. Mean modal doses were 3.8 mg/day (SD=1.8) of risperidone and 6.2 mg/day (SD=2.9) of haloperidol. Significantly greater reductions in Young Mania Rating Scale scores at endpoint and over time were seen in the risperidone group and in the haloperidol group, compared with the placebo group. Young Mania Rating Scale total scores improved with risperidone and with haloperidol both in patients with psychotic features and in those without psychotic features at baseline. Extrapyramidal Symptom Rating Scale total scores at endpoint were significantly higher in the haloperidol patients than in the placebo patients. Antiparkinsonian medications were received by 8%, 17%, and 38% of patients in the placebo, risperidone, and haloperidol groups, respectively. Risperidone plus a mood stabilizer was more efficacious than a mood stabilizer alone, and as efficacious as haloperidol plus a mood stabilizer, for the rapid control of manic symptoms and was well tolerated.


Zebrafish cdc6 hypomorphic mutation causes Meier-Gorlin syndrome-like phenotype.
Cell Division Cycle 6 (Cdc6) is a component of pre-replicative complex (preRC) forming on DNA replication origins in eukaryotes. Recessive mutations in ORC1, ORC4, ORC6, CDT1 or CDC6 of the preRC in human cause Meier-Gorlin syndrome (MGS) that is characterized by impaired post-natal growth, short stature and microcephaly. However, vertebrate models of MGS have not been reported. Through N-ethyl-N-nitrosourea mutagenesis and Cas9 knockout, we generate several cdc6 mutant lines in zebrafish. Loss-of-function mutations of cdc6, as manifested by cdc6tsu4305 and cdc6tsu7cd mutants, lead to embryonic lethality due to cell cycle arrest at the S phase and extensive apoptosis. Embryos homozygous for a cdc6 hypomorphic mutation, cdc6tsu21cd, develop normally during embryogenesis. Later on, compared with their wild-type (WT) siblings, cdc6tsu21cd mutant fish show growth retardation, and their body weight and length in adulthood are greatly reduced, which resemble human MGS. Surprisingly, cdc6tsu21cd mutant fish become males with a short life and fail to mate with WT females, suggesting defective reproduction. Overexpression of Cdc6 mutant forms, which mimic human CDC6(T323R) mutation found in a MGS patient, in zebrafish cdc6tsu4305 mutant embryos partially represses cell death phenotype, suggesting that the human CDC6(T323R) mutation is a hypomorph. cdc6tsu21cd mutant fish will be useful to detect more tissue defects and develop medical treatment strategies for MGS patients.


Dementia in patients hospitalized with stroke: rates, time course, and clinico-pathologic factors.
Stroke is a risk factor for dementia and dementia predisposes to stroke. Dementia prevalence in subjects with stroke is comparable with that seen in stroke-free subjects who are 10 years older. Although overall there is heterogeneity between studies of prestroke and poststroke dementia, stratifying by study method and clinical criteria (e.g. inclusion/exclusion of prestroke dementia, first ever vs. any vs. recurrent stroke) results in reasonably consistent estimates. Pooled dementia rates from studies of consecutive patients hospitalized with stroke indicate that around 10% have dementia prior to first stroke and 10% have new dementia in the first year after first-ever stroke with highest rates (over 30%) seen after recurrent stroke. After the immediate high-risk poststroke period, rates of new incident dementia are lower but remain elevated at around four times the background risk. Factors associated with prestroke and poststroke dementia are broadly similar but age, medial temporal lobe atrophy, female sex, and family history are more strongly associated with prestroke dementia suggesting a greater role for degenerative pathology. Poststroke dementia is associated with factors indicating a reduced cognitive reserve (prestroke cognitive decline, premorbid disability, low education, white matter disease, and atrophy) and is also strongly associated with stroke factors (lesion size, multiple lesions, and stroke recurrence) and complications of stroke (delirium, seizures, hypotension, systemic illness and incontinence) indicating the likely impact of optimal acute stroke care and secondary prevention in reducing the burden of dementia. Future studies are needed to clarify the interaction between degenerative, vascular, and systemic processes in the development of stroke-associated dementia.


Color discrimination thresholds in a cichlid fish: Metriaclima benetos.
Color vision is essential for animals as it allows them to detect, recognize and discriminate between colored objects. Studies analyzing color vision require an integrative approach, combining behavioral experiments, physiological models and quantitative analyses of photoreceptor stimulation. Here, we demonstrate, for the first time, the limits of chromatic discrimination in Metriaclima benetos, a rock-dwelling cichlid from Lake Malawi, using behavioral experiments and visual modeling. Fish were trained to discriminate between colored stimuli. Color discrimination thresholds were quantified by testing fish chromatic discrimination between the rewarded stimulus and distracter stimuli that varied in chromatic distance (ΔS). This was done under fluorescent lights alone and with additional violet lights. Our results provide two main outcomes. First, cichlid color discrimination thresholds correspond with predictions from the receptor noise limited (RNL) model but only if we assume a Weber fraction higher than the typical value of 5%. Second, cichlids may exhibit limited color constancy under certain lighting conditions as most individuals failed to discriminate colors when violet light was added. We further used the color discrimination thresholds obtained from these experiments to model color discrimination of actual fish colors and backgrounds under natural lighting for Lake Malawi. We found that, for M. benetos, blue is most chromatically contrasting against yellows and space-light, which might be important for discriminating male nuptial colorations and detecting males against the background. This study highlights the importance of lab-based behavioral experiments in understanding color vision and in parameterizing the assumptions of the RNL vision model for different species.


Atypical chest pain in the elderly.
It has been shown that the elderly, and certain other groups, may have atypical clinical presentations of acute MI. It is important for the clinician to educate patients about the common atypical symptoms that may be experienced with an MI, such as dyspnea, fatigue, nausea, vomiting, and syncope. The clinician must always rule out acute MI in any patient who presents with these symptoms, or who presents with falls, sudden weakness, or worsening CHF. In order to treat patients aggressively and with the greatest benefit (i.e. thrombolytics or other reperfusion therapy), we must teach our patients and ourselves to recognize "silent" MIs. This will decrease the morbidity and mortality rates of acute MI in the elderly.


Microvesiculation and disease.
The important roles of extracellular vesicles in the pathogenesis of various diseases are rapidly being elucidated. As important vehicles of intercellular communication, extracellular vesicles, which comprise microvesicles and exosomes, are revealing important roles in cancer tumorigenesis and metastases and in the spread of infectious disease. The September 2012 Focused Meeting 'Microvesiculation and Disease' brought together researchers working on extracellular vesicles. The papers in this issue of Biochemical Society Transactions review work in areas including HIV infection, kidney disease, hypoxia-mediated tumorigenesis and down-regulation of immune cell functions in acute myeloid leukaemia by tumour-derived exosomes. In all cases, microvesicles and exosomes have been demonstrated to be important factors leading to the pathophysiology of disease or indeed as therapeutic vehicles in possible new treatments. The aim was, having enhanced our molecular understanding of the contribution of microvesicles and exosomes to disease in vitro, to begin to apply this knowledge to in vivo models of disease.


Ultrasonography of the optic nerve in neurocritically ill patients.
The rapid diagnosis of intracranial hypertension is urgently needed for therapeutic reasons in various clinical settings. This can rarely be achieved without invasive procedures such as intracranial pressure (ICP) monitoring or neuroimaging. The optic nerve is surrounded by cerebrospinal fluid (CSF) and dura mater, which forms the optic nerve sheath (ONS). Because of the connection with the intracranial subarachnoid space, ONS diameter (ONSD) is influenced by CSF pressure variations. Bedside ultrasonographic measurement of ONSD has been proposed as a non-invasive and reliable means to detect raised ICP in neurocritically ill patients. In several studies, it proves to have a good correlation with the direct measurement of ICP and a low interobserver variability. However, no general consensus exists over the upper normal ONSD limit. We performed a review of the literature on the use of the ultrasonography of the optic nerve in the evaluation of patients with suspected intracranial hypertension. The aim of this review is to describe the technique and to assess the validity of this diagnostic method.


p16 and K-ras gene mutations in the intraductal precursors of human pancreatic adenocarcinoma.
Pancreatic adenocarcinoma is thought to arise from a noninvasive neoplastic precursor, the pancreatic intraductal lesion (PIL). Mutations of the K-ras gene are known to occur in PILs, but their high prevalence among PILs within the general population probably limit the use of K-ras as a marker of eventual clinical risk. In search of genetic constellations that might indicate the progression of some PILs toward an invasive phenotype, mutations at both the K-ras and p16 genes were sought within PILs of 10 pancreata resected for adenocarcinoma. K-ras mutations were present in most PILs and in nearly all PILs having nuclear atypia. In half of the patients, two or more unique K-ras mutations were identified among distinct PILs, which is evidence for the separate clonal evolution of multiple pancreatic neoplasms within individual patients. p16 alterations (one homozygous deletion and three point mutations) were found in 4 of the 10 carcinomas; these four pancreata harbored p16 alterations in three of nine PILs, of which one was a "histologically early" lesion. Two patients had p16 alterations in PILs matching those of the associated carcinomas. p16 mutations were not found in PILs of pancreata having wild-type p16 in the carcinoma, nor were they found in ducts having normal histology. It is suggested that alterations of the p16 gene affect a subset of PILs that contain mutations of the K-ras gene and that these mutations might identify high-risk precursors of the invasive malignancy.


Poly(ADP-ribose) synthetase associated to cytoplasmic structures of meiotic cells.
An active poly(ADP-ribose) synthetase has been found to be associated to mitochondria of frog oocytes and of testis from various species. The enzyme was also found in association with the microsomal-ribosomes fraction of the testis. Both enzymes are strongly inhibited by nicotinamide, thymidine, and theophylline, and they synthesize oligomers of ADP-ribose of an average chain length of 2-6 residues. The apparent km for NAD+ of the mitochondrial enzyme is 22 micro M which contrasts with the corresponding value of the chromatin-bound enzyme, 210 micro M NAD+. Treatment of the isolated mitochondria with Triton X-100 revealed that the enzyme is associated to a polydisperse structure. The higher specific activity of the enzyme associated to mitochondria was found in testis of 20 to 30 days old rats. This development study suggest that the maximal activity of the enzyme is associated to mitochondria belonging to primary spermatocytes. Poly(ADP-ribose) synthetase was also found in association of the microsome-ribosome fraction of rat, mouse, carp, and bull testis. This activity is 20 to 30 times higher than in the same fraction of the liver. Contrary to the chromatin-bound enzyme, the microsome-ribosome enzyme activity is stimulated 3 to 4 times by DNA, but it is slightly inhibited by histone H1. The enzyme seems to be associated to ribonucleoproteins of a very polydisperse nature. The possible relationship of poly(ADP-ribose) synthetase with the intermitochondrial cement and with the chromatoid body will be discussed. These are very unique structures only present in germinal cells.


Composition and seasonality of an amphipod community associated to the algae Bryocladia trysigera.
The monthly fluctuations of amphipods associated to the algae Bryocladia trysigera was described from March 1997 to February 1998 at the rocky shore known as Poço de Anchieta in the Peruíbe Beach, Itanhaém, Southeastern Brazil. A total of 75,344 individuals were sampled, belonging to 10 species and 9 families. Three species dominated the phytal in number: Hyale nigra, Caprella danileviskii and Caprella penantis. Despite the alternation in dominance of the 3 most abundant species, the amphipod species composition remained generally unchanged, so that the majority of the species were observed in all sampling months. The temporally changing pattern of the community structure, with a decline in amphipod abundance in winter followed by an increase in spring, was probably due to a higher predation pressure in winter period.


d-Glucose Adsorption on the TiO2 Anatase (100) Surface: A Direct Comparison Between Cluster-Based and Periodic Approaches.
Titanium dioxide (TiO2) has been extensively studied as a suitable material for a wide range of fields including catalysis and sensing. For example, TiO2-based nanoparticles are active in the catalytic conversion of glucose into value-added chemicals, while the good biocompatibility of titania allows for its application in innovative biosensing devices for glucose detection. A key process for efficient and selective biosensors and catalysts is the interaction and binding mode between the analyte and the sensor/catalyst surface. The relevant features regard both the molecular recognition event and its effects on the nanoparticle electronic structure. In this work, we address both these features by combining two first-principles methods based on periodic boundary conditions and cluster approaches (CAs). While the former allows for the investigation of extended materials and surfaces, CAs focus only on a local region of the surface but allow for using hybrid functionals with low computational cost, leading to a highly accurate description of electronic properties. Moreover, the CA is suitable for the study of reaction mechanisms and charged systems, which can be cumbersome with PBC. Here, a direct and detailed comparison of the two computational methodologies is applied for the investigation of d-glucose on the TiO2 (100) anatase surface. As an alternative to the commonly used PBC calculations, the CA is successfully exploited to characterize the formation of surface and subsurface oxygen vacancies and to determine their decisive role in d-glucose adsorption. The results of such direct comparison allow for the selection of an efficient, finite-size structural model that is suitable for future investigations of biosensor electrocatalytic processes and biomass conversion catalysis.


Therapeutic effects of vasoactive intestinal peptide in the trinitrobenzene sulfonic acid mice model of Crohn's disease.
Crohn's disease (CD) is a chronic debilitating disease of unknown etiology that is characterized by severe inflammation of the colon. Vasoactive intestinal peptide (VIP) has recently emerged as a promising candidate for treatment of inflammatory Th1-driven diseases. We studied the effect of VIP in trinitrobenzene sulfonic acid (TNBS)-induced colitis, which has clinical and molecular features in common with CD. A 3-mg enema of TNBS was given to BALB/c mice, and VIP (1 nmol) was given either as a single dose at 12 hours or every other day. Weight loss, histopathology, and chemokine and cytokine levels in serum and colon extracts were assessed. VIP was also tested given 5 days after the onset of TNBS-induced colitis, and its effect was analyzed given a second dose of TNBS. Treatment with VIP reduced the clinical and histopathologic severity of TNBS-induced colitis, abrogating body weight loss, diarrhea, and macroscopic and microscopic intestinal inflammation. The therapeutic effects of VIP were associated with down-regulation of both inflammatory and Th1-driven autoimmune responses, including tumor necrosis factor alpha, interleukin 1, and interleukin 6 in colon extracts and serum as well as interferon gamma by splenic and lamina propria CD4(+) T cells. VIP reduced disease severity when given after disease onset and dramatically reduced disease recurrence given a second dose of TNBS. Our data suggest that VIP has beneficial prophylactic and therapeutic effects in TNBS-induced colitis and is a promising candidate to test for potential benefits in CD.


Early detection of fronto-temporal degeneration by clinical evaluation.
Although Alzheimer's Disease (AD) is the most common degenerative dementia, other neuropathological processes are well known to cause dementia syndromes. Fronto-temporal degeneration (FTD) is one such entity, and often produces a clinical presentation distinct from that of AD. Some FTD cases are shown at autopsy to be classic Pick's Disease, but others defy precise classification at this point because they lack characteristic Pick bodies. We describe a case of putative FTD in a 54-year-old man with personality change and complete Klüver-Bucy syndrome. Neuropsychological evaluation disclosed evidence of extensive frontal system dysfunction, with lesser problems in memory, language, and visuospatial skills. Magnetic resonance imaging studies after 17 months of the illness were largely nonspecific, but a follow-up scan 16 months later revealed bifrontal and bitemporal atrophy with ventricular enlargement. Neuromorphometric analysis suggested an increase in cortical atrophy and ventricular dilation over time. This case emphasizes the value of careful clinical evaluation in unusual non-AD degenerative dementias and suggests that neuroimaging studies may be less sensitive in the early diagnosis of such cases.


Practical guidelines for adaptive seamless phase II/III clinical trials that use Bayesian methods.
Hommel (Biometrical Journal; 45:581-589) proposed a flexible testing procedure for seamless phase II/III clinical trials. Schmidli et al. (Statistics in Medicine; 26:4925-4938), Kimani et al. (Statistics in Medicine; 28:917-936) and Brannath et al. (Statistics in Medicine; 28:1445-1463) exploited the flexible testing of Hommel to propose adaptation in seamless phase II/III clinical trials that incorporate prior knowledge by using Bayesian methods. In this paper, we show that adaptation incorporating prior knowledge may lead to higher power. Other important issues to consider in such adaptive designs are the gain in power (or saving in patients) over traditional testing and how utility values used to make the adaptation may be used to stop a trial early. In contrast to the aforementioned authors, we discuss these issues in detail and propose a unified approach to address them so that implementing the aforementioned designs and proposing similar designs is clearer.


Long-Term Expansion in Platelet Lysate Increases Growth of Peripheral Blood-Derived Endothelial-Colony Forming Cells and Their Growth Factor-Induced Sprouting Capacity.
Efficient implementation of peripheral blood-derived endothelial-colony cells (PB-ECFCs) as a therapeutical tool requires isolation and generation of a sufficient number of cells in ex vivo conditions devoid of animal-derived products. At present, little is known how the isolation and expansion procedure in xenogeneic-free conditions affects the therapeutical capacity of PB-ECFCs. The findings presented in this study indicate that human platelet lysate (PL) as a serum substitute yields twice more colonies per mL blood compared to the conventional isolation with fetal bovine serum (FBS). Isolated ECFCs displayed a higher proliferative ability in PL supplemented medium than cells in FBS medium during 30 days expansion. The cells at 18 cumulative population doubling levels (CPDL) retained their proliferative capacity, showed higher sprouting ability in fibrin matrices upon stimulation with FGF-2 and VEGF-A than the cells at 6 CPDL, and displayed low β-galactosidase activity. The increased sprouting of PB-ECFCs at 18 CPDL was accompanied by an intrinsic activation of the uPA/uPAR fibrinolytic system. Induced deficiency of uPA (urokinase-type plasminogen activator) or uPAR (uPA receptor) by siRNA technology completely abolished the angiogenic ability of PB-ECFCs in fibrin matrices. During the serial expansion, the gene induction of the markers associated with inflammatory activation such as VCAM-1 and ICAM-1 did not occur or only to limited extent. While further propagation up to 31 CPDL proceeded at a comparable rate, a marked upregulation of inflammatory markers occurred in all donors accompanied by a further increase of uPA/uPAR gene induction. The observed induction of inflammatory genes at later stages of long-term propagation of PB-ECFCs underpins the necessity to determine the right time-point for harvesting of sufficient number of cells with preserved therapeutical potential. The presented isolation method and subsequent cell expansion in platelet lysate supplemented culture medium permits suitable large-scale propagation of PB-ECFC. For optimal use of PB-ECFCs in clinical settings, our data suggest that 15-20 CPDL is the most adequate maturation stage.


Marrubium persicum İmproved the Biological Parameters Associated with Angiogenesis and İnflammation in Mice.
The genus Marrubium, belonging to the family Lamiaceae, is highly praised in herbal medicine of different countries for having renowned healing properties. Herein, the anti-inflammatory and anti-angiogenesis potential of Marrubium persicum methanol extract was evaluated in a mouse air pouch model of inflammation. Aerial parts of M. persicum were solvent extracted using the Soxhlet apparatus. Subsequently, air injections were performed (for 3 days) into the mice's backs to bring about an air pouch, while carrageenan was used to induce inflammation. The mice were divided into four groups, including; negative control (normal saline into the pouch), control (carrageenan), treatment and positive control (dexamethasone). The inflammatory markers were analyzed 48h after injecting carrageenan, and a haemoglobin assay kit assessed the quantification of angiogenesis in granulation tissue. M. persicum methanol extract at doses of 3.5, 5, 7.5 and 10 mg/kg represented significant decreases in inflammatory parameters. Compared to the control group, the optimum dose (3.5 mg/kg) lessened the myeloperoxidase (MPO) and angiogenesis activity, as well as haemoglobin levels. In sum, the methanol extract of M. persicum exhibited anti-inflammatory effects against carrageenan-induced inflammation, which could be related to its antioxidant and inhibitory effects on neutrophils' infiltration.


Comorbidities in multiple myeloma and implications on survival: A population-based study.
High proportion of patients with multiple myeloma suffer from comorbidities which may alter clinical management. Therefore, our aims were to evaluate the prevalence of comorbidities and their impact on survival. We included patients diagnosed with multiple myeloma 1990-2013 in Sweden and all diagnoses from each patient from 1985. A total of 13 656 patients with multiple myeloma were included in the study, thereof 7404 (54%) had comorbidity at diagnosis. The risk of death was increased for those with one comorbidity at diagnosis compared to those without any comorbidity (hazard ratio = 1.19; 95% confidence interval:1.14-1.25); this risk was higher for those with two (1.38; 1.30-1.47) and three or more comorbidities (1.72; 1.62-1.83). Furthermore, the risk of death was increased in patients with prior history of cancer, arrhythmia, heart failure, diabetes mellitus, cerebrovascular disease, chronic lung disease, psychological disease, peptic ulcer, neurological disease, peripheral vascular disease, chronic kidney disease, dementia, and inflammatory bowel disease. This large study shows that over 50% of multiple myeloma patients have a comorbidity at diagnosis and survival decreased with increasing numbers of comorbidities. This emphasizes the importance of comorbidities when evaluating patients and deciding on treatment strategies for individuals with multiple myeloma.


Pioneer re-entry device for iliac chronic total occlusion: truly a paradigm shift.
The number of percutaneous revascularization procedures performed for symptomatic peripheral arterial disease has significantly increased over the past several years. Traditionally, the use of percutaneous techniques were limited to certain anatomic subsets, such as stenosis or focal occlusions, with surgical treatment preferred for more extensive disease. More recently, endovascular specialists are facing the challenge of peripheral chronic total occlusions. Furthermore, unlike the coronary circulation, these occlusions are often very long and associated with other features of complexity such as severe calcifications. One of the primary issues concerning these lesions is the ability to safely achieve initial angiographic success. This article focus indeed on the Pioneer catheter, a new lumen re-entry device exploiting intravascular ultrasound imaging that was used in a case of totally occluded left common iliac artery with favorable results. The details of this technique, and how this catheter helped in re-entering the true lumen at the aortic bifurcation after subintimal dissection, are thoroughly discussed.


On the development of ectopic eyes in Drosophila melanogaster produced by the mutation extra eye (ee).
The mutation ee often produces an ectopic eye on the vertex that is a mirror image partial duplication of the normal eye on the ipsilateral side of the head. The pattern of the duplication and a clonal analysis by mitotic recombination indicate that the duplications are of dorsal eye and orbital structures. Large ectopic eyes (more than 100 ommatidia) and their surrounding bristles may be produced without cuticular deficiencies. The penetrance of ee is temperature dependent with penetrance higher (72%) at 25 degrees and 29 degrees than at 19 degrees (43%). Temperature shift experiments show two temperature-sensitive periods: one at midembryogenesis, the other at mid-first larval instar. Microscopic examination of ee late-second and third instar imaginal cephalic discs show no indication of growth of the extra tissue needed to produce the duplication until after mid-third instar. This was confirmed by cell counts of ee and wild-type discs. There is no evidence of differential cell death in the two types of discs at this stage, although much earlier cell death is postulated. Tests for cell autonomy of the mutation by the production of morphogenetic clones suggest nonautonomy. Formation of pattern duplications by mutant genes is discussed in terms of cell death that eliminates whole developmental compartments, restricted cell death that occurs within a compartment, extensive cell death within a compartment and proliferative growth unassociated with cell lethality.


A let-7 microRNA-binding site polymorphism in 3'-untranslated region of KRAS gene predicts response in wild-type KRAS patients with metastatic colorectal cancer treated with cetuximab monotherapy.
recent studies have found that KRAS mutations predict resistance to monoclonal antibodies targeting the epidermal growth factor receptor in metastatic colorectal cancer (mCRC). A polymorphism in a let-7 microRNA complementary site (lcs6) in the KRAS 3' untranslated region (UTR) is associated with an increased cancer risk in non-small-cell lung cancer and reduced overall survival (OS) in oral cancers. We tested the hypothesis whether this polymorphism may be associated with clinical outcome in KRAS wild-type (KRASwt) mCRC patients treated with cetuximab monotherapy. the presence of KRAS let-7 lcs6 polymorphism was evaluated in 130 mCRC patients who were enrolled in a phase II study of cetuximab monotherapy (IMCL-0144). Genomic DNA was extracted from dissected formalin-fixed paraffin-embedded tumor tissue, KRAS mutation status and polymorphism were assessed using direct sequencing and PCR restriction fragment length polymorphism technique. KRAS let-7 lcs6 polymorphism was found to be related to object response rate (ORR) in mCRC patients whose tumors had KRASwt. The 12 KRASwt patients harboring at least a variant G allele (TG or GG) had a 42% ORR compared with a 9% ORR in 55 KRASwt patients with let-7 lcs6 TT genotype (P = 0.02, Fisher's exact test). KRASwt patients with TG/GG genotypes had trend of longer median progression-free survival (3.9 versus 1.3 months) and OS (10.7 versus 6.4 months) compared to those with TT genotypes. these results are the first to indicate that the KRAS 3'UTR polymorphism may predict for cetuximab responsiveness in KRASwt mCRC patients, which warrants validation in other clinical trials.


Process-based expansion and neural differentiation of human pluripotent stem cells for transplantation and disease modeling.
Robust strategies for developing patient-specific, human, induced pluripotent stem cell (iPSC)-based therapies of the brain require an ability to derive large numbers of highly defined neural cells. Recent progress in iPSC culture techniques includes partial-to-complete elimination of feeder layers, use of defined media, and single-cell passaging. However, these techniques still require embryoid body formation or coculture for differentiation into neural stem cells (NSCs). In addition, none of the published methodologies has employed all of the advances in a single culture system. Here we describe a reliable method for long-term, single-cell passaging of PSCs using a feeder-free, defined culture system that produces confluent, adherent PSCs that can be differentiated into NSCs. To provide a basis for robust quality control, we have devised a system of cellular nomenclature that describes an accurate genotype and phenotype of the cells at specific stages in the process. We demonstrate that this protocol allows for the efficient, large-scale, cGMP-compliant production of transplantable NSCs from all lines tested. We also show that NSCs generated from iPSCs produced with the process described are capable of forming both glia defined by their expression of S100β and neurons that fire repetitive action potentials.


Velocardiofacial syndrome with facial and pinna asymmetries.
This study reports some of the most important clinical features of the velocardiofacial syndrome (hypoplastic zygomatic arch, prominent nose with square nasal root, bilateral epicanthus, downslanting palpebral fissures, and learning disabilities) in a Brazilian boy presenting face and pinna asymmetries. These findings may facilitate the diagnosis of this syndrome.


The neural basis of perceptual category learning in human infants.
We measured looking times and ERPs to examine the cognitive and brain bases of perceptual category learning in 6-month-old infants. In Experiment 1, we showed that categorization and exemplar discrimination rely on different cortical processes. Specifically, the repetition of individual exemplars resulted in differential cortical processing at posterior channels at an early stage during object processing (100-300 msec), whereas discriminating among members of different categories was reflected in ERP differences over anterior cortical regions occurring later in time (300-500 msec) than the repetition effects. In Experiment 2, replicating the findings of Study 1, we found that infants engage the same cortical processes to categorize visual objects into basic-level categories, regardless of whether a basic (bird vs. fish) or global level is crossed (birds vs. cars). This pattern of findings is consistent with perceptual accounts of infant categorization [Quinn, P. C., & Eimas, P. D. Perceptual organization and categorization in young infants. In C. Rovee-Collier & L. P. Lipsitt (Eds.), Advances in infancy research ( pp. 1-36). Norwood, NJ: Ablex, 1996] and accords with recent adult neural-level models of perceptual categorization.


Widespread brain reorganization perturbs visuomotor coordination in early glaucoma.
Glaucoma is the world's leading cause of irreversible blindness, and falls are a major public health concern in glaucoma patients. Although recent evidence suggests the involvements of the brain toward advanced glaucoma stages, the early brain changes and their clinical and behavioral consequences remain poorly described. This study aims to determine how glaucoma may impair the brain structurally and functionally within and beyond the visual pathway in the early stages, and whether these changes can explain visuomotor impairments in glaucoma. Using multi-parametric magnetic resonance imaging, glaucoma patients presented compromised white matter integrity along the central visual pathway and around the supramarginal gyrus, as well as reduced functional connectivity between the supramarginal gyrus and the visual occipital and superior sensorimotor areas when compared to healthy controls. Furthermore, decreased functional connectivity between the supramarginal gyrus and the visual brain network may negatively impact postural control measured with dynamic posturography in glaucoma patients. Taken together, this study demonstrates that widespread structural and functional brain reorganization is taking place in areas associated with visuomotor coordination in early glaucoma. These results implicate an important central mechanism by which glaucoma patients may be susceptible to visual impairments and increased risk of falls.


Co-occuring psychiatric symptoms in children perinatally infected with HIV and peer comparison sample.
To compare the rates of psychopathology in youths perinatally infected with HIV (N = 319) with a comparison sample of peers (N = 256) either HIV-exposed or living in households with HIV-infected family members. Participants were randomly recruited from 29 sites in the United States and Puerto Rico and completed an extensive battery of measures including standardized DSM-IV-referenced ratings scales. The HIV+ group was relatively healthy (73% with CD4% >25%), and 92% were actively receiving antiretroviral therapy. Youths with HIV (17%) met symptom and impairment criteria for the following disorders: attention-deficit/hyperactivity disorder (12%), oppositional defiant disorder (5%), conduct disorder (1%), generalized anxiety disorder (2%), separation anxiety disorder (1%), depressive disorder (2%), or manic episode (1%). Many youths with HIV (27%) and peers (26%) were rated (either self- or caregiver report) as having psychiatric problems that interfered with academic or social functioning. With the exception of somatization disorder, the HIV+ group did not evidence higher rates or severity of psychopathology than peers, although rates for both groups were higher than the general population. Nevertheless, self-awareness of HIV infection in younger children was associated with more severe symptomatology, and youths with HIV had higher lifetime rates of special education (44 vs 32%), psychopharmacological (23 vs 12%), or behavioral (27 vs 17%) interventions. Youth-caregiver agreement was modest, and youths reported more impairment. HIV infection was not associated with differentially greater levels of current psychopathology; nevertheless, investigation of relations with developmental changes and specific illness parameters and treatments are ongoing.


Direct Current Stimulation Modulates Synaptic Facilitation via Distinct Presynaptic Calcium Channels.
Transcranial direct current stimulation (tDCS) is a subthreshold neurostimulation technique known for ameliorating neuropsychiatric conditions. The principal mechanism of tDCS is the differential polarization of subcellular neuronal compartments, particularly the axon terminals that are sensitive to external electrical fields. Yet, the underlying mechanism of tDCS is not fully clear. Here, we hypothesized that direct current stimulation (DCS)-induced modulation of presynaptic calcium channel conductance alters axon terminal dynamics with regard to synaptic vesicle release. To examine the involvement of calcium-channel subtypes in tDCS, we recorded spontaneous excitatory postsynaptic currents (sEPSCs) from cortical layer-V pyramidal neurons under DCS while selectively inhibiting distinct subtypes of voltage-dependent calcium channels. Blocking P/Q or N-type calcium channels occluded the effects of DCS on sEPSCs, demonstrating their critical role in the process of DCS-induced modulation of spontaneous vesicle release. However, inhibiting T-type calcium channels did not occlude DCS-induced modulation of sEPSCs, suggesting that despite being active in the subthreshold range, T-type calcium channels are not involved in the axonal effects of DCS. DCS modulates synaptic facilitation by regulating calcium channels in axon terminals, primarily via controlling P/Q and N-type calcium channels, while T-type calcium channels are not involved in this mechanism.


[The role of stereotactic 11G vacuum biopsy for clarification of BI-RADS IV findings in mammography].
To evaluate the potential of stereotactic vacuum breast biopsy in the histologic evaluation of suspicious mammographic findings ( BI-RADS IV). In 221 patients with 227 probable mammographic lesions categorized as ( BI-RADS -IV), stereotactic biopsies were performed with an 11-gauge vacuum-assisted biopsy device (Mammotome). The evaluation included the histology of the specimens obtained with the Mammotome or with surgery, the time for the biopsy, the amount of bleeding, number of rotations and procured specimens, the extent of the resection and the complications. The biopsies were technically successful in 214 of the 227 probable mammographic lesions, with 176 lesions mostly resected and 34 lesions removed more than 50 %. No representative tissue was obtained from 4 lesions. All biopsies were performed without any clinically relevant complications and terminated after adequate material was obtained (O 28 specimens, 2.58 rotations). The mean time needed for performing the biopsy was 40.2 minutes. The histologic findings were DCIS (42 lesions), ADH (7 lesions), LCIS (3 lesions), ID-Ca (14 lesions, IL-Ca (3 lesions), and IDL-Ca (1 lesion). In 28 of 42 lesions with the initial DCIS histology, the surgical histology was also DCIS (n=28) or no residual tumor (n=10). In 4 lesions with an initial DCIS-histology, the surgical histology was invasive ductal cancer (9.5 %). The late follow-up examinations (up to 3 years) did not find any evidence of a false negative biopsy. Stereotactic vacuum breast biopsy ideally complements existing breast biopsy methods. The method is minimal invasive with a low rate of mostly minor complications.


Natural history and clinical biomarkers of progression in X-linked retinitis pigmentosa: a systematic review.
X-linked retinitis pigmentosa (XLRP) accounts for a significant proportion of certifiable blindness in working-age adults. The objectives of this study were to: (1) synthesize the best available evidence regarding the natural history of disease progression and (2) identify the best current clinical biomarkers for monitoring disease progression, which will be important in planned gene therapy trials for this condition. Patient population: XLRP affected males. Main outcomes: A systematic review of the literature was undertaken with data sought on overall annual progression for clinical biomarkers using optical coherence tomography (OCT), fundus autofluorescence (FAF), visual acuity, electroretinography and visual fields. To assess which outcome was best for monitoring progression, data on reliability, interocular correlation and structure-function correlation were extracted. A total of 17 studies met the inclusion criteria. Studies estimated progression at between 4% to 19% per year with longitudinal data. Where an overall model was produced with cross-sectional data, the trend was usually best fit by a logarithmic function with an annual exponential decline rate between 4.7% and 8.0%. The evidence suggested the ellipsoid zone (EZ) width on OCT and outer ring area (ORA) on FAF as the most useful biomarkers having excellent interocular symmetry, reproducibility and functional correlation. Using different clinical biomarkers, XLRP progresses at a rate of 4 to 19% per year. Ellipsoid zone (EZ) width and ORA are the most robust biomarkers with the potential to be used in trials where one eye serves as a control for the other.


The effects of stimulus variability on the perceptual learning of speech and non-speech stimuli.
Previous studies suggest fundamental differences between the perceptual learning of speech and non-speech stimuli. One major difference is in the way variability in the training set affects learning and its generalization to untrained stimuli: training-set variability appears to facilitate speech learning, while slowing or altogether extinguishing non-speech auditory learning. We asked whether the reason for this apparent difference is a consequence of the very different methodologies used in speech and non-speech studies. We hypothesized that speech and non-speech training would result in a similar pattern of learning if they were trained using the same training regimen. We used a 2 (random vs. blocked pre- and post-testing) × 2 (random vs. blocked training) × 2 (speech vs. non-speech discrimination task) study design, yielding 8 training groups. A further 2 groups acted as untrained controls, tested with either random or blocked stimuli. The speech task required syllable discrimination along 4 minimal-pair continua (e.g., bee-dee), and the non-speech stimuli required duration discrimination around 4 base durations (e.g., 50 ms). Training and testing required listeners to pick the odd-one-out of three stimuli, two of which were the base duration or phoneme continuum endpoint and the third varied adaptively. Training was administered in 9 sessions of 640 trials each, spread over 4-8 weeks. Significant learning was only observed following speech training, with similar learning rates and full generalization regardless of whether training used random or blocked schedules. No learning was observed for duration discrimination with either training regimen. We therefore conclude that the two stimulus classes respond differently to the same training regimen. A reasonable interpretation of the findings is that speech is perceived categorically, enabling learning in either paradigm, while the different base durations are not well-enough differentiated to allow for categorization, resulting in disruption to learning.


The influence of pause, attack, and decay duration of the ongoing envelope on sound lateralization.
Klein-Hennig et al. [J. Acoust. Soc. Am. 129, 3856-3872 (2011)] introduced a class of high-frequency stimuli for which the envelope shape can be altered by independently varying the attack, hold, decay, and pause durations. These stimuli, originally employed for testing the shape dependence of human listeners' sensitivity to interaural temporal differences (ITDs) in the ongoing envelope, were used to measure the lateralization produced by fixed interaural disparities. Consistent with the threshold ITD data, a steep attack and a non-zero pause facilitate strong ITD-based lateralization. In contrast, those conditions resulted in the smallest interaural level-based lateralization.


Functional and phylogenetic relatedness in temporary wetland invertebrates: current macroecological patterns and implications for future climatic change scenarios.
In freshwater ecosystems, species compositions are known to be determined hierarchically by large to small‑scale environmental factors, based on the biological traits of the organisms. However, in ephemeral habitats this heuristic framework remains largely untested. Although temporary wetland faunas are constrained by a local filter (i.e., desiccation), we propose its magnitude may still depend on large-scale climate characteristics. If this is true, climate should be related to the degree of functional and taxonomic relatedness of invertebrate communities inhabiting seasonal wetlands. We tested this hypothesis in two ways. First, based on 52 biological traits for invertebrates, we conducted a case study to explore functional trends among temperate seasonal wetlands differing in the harshness (i.e., dryness) of their dry season. After finding evidence of trait filtering, we addressed whether it could be generalized across a broader climatic scale. To this end, a meta-analysis (225 seasonal wetlands spread across broad climatic categories: Arid, Temperate, and Cold) allowed us to identify whether an equivalent climate-dependent pattern of trait richness was consistent between the Nearctic and the Western Palearctic. Functional overlap of invertebrates increased from mild (i.e., Temperate) to harsher climates (i.e., Arid and Cold), and phylogenetic clustering (using taxonomy as a surrogate) was highest in Arid and lowest in Temperate wetlands. We show that, (i) as has been described in streams, higher relatedness than would be expected by chance is generally observed in seasonal wetland invertebrate communities; and (ii) this relatedness is not constant but climate-dependent, with the climate under which a given seasonal wetland is located determining the functional overlap and the phylogenetic clustering of the community. Finally, using a space-for-time substitution approach we suggest our results may anticipate how the invertebrate biodiversity embedded in these vulnerable and often overlooked ecosystems will be affected by long-term climate change.


Right ventricular oxygen delivery as a determinant of right ventricular functional reserve during exercise in juvenile swine with chronic pulmonary hypertension.
Assessing right ventricular (RV) functional reserve is important for determining clinical status and prognosis in patients with pulmonary hypertension (PH). In this study, we aimed to establish RV oxygen (O2) delivery as a determinant for RV functional reserve during exercise in swine with chronic PH. Chronic PH was induced by pulmonary vein banding (PVB), with sham operation serving as control. RV function and RV O2 delivery were measured over time in chronically instrumented swine, up to 12 wk after PVB at rest and during exercise. At rest, RV afterload (pulmonary artery pressure and arterial elastance) and contractility (Ees and dP/dtmax) were higher in PH compared with control with preserved cardiac index and RV O2 delivery. However, RV functional reserve, as measured by the exercise-induced relative change (Δ) in cardiac index, dP/dtmax, and end-systolic elastance (Ees), was decreased in PH, and RV pulmonary arterial coupling was lower both at rest and during exercise in PH. Furthermore, the increase in RV O2 delivery was attenuated in PH during exercise principally due to a lower systolic coronary blood flow in combination with an attenuated increase in aorta pressure while arterial O2 content was not significantly altered in PH. Moreover, RV O2 delivery reserve correlated with RV functional reserve, Δcardiac index (r2 = 0.85), ΔdP/dtmax (r2 = 0.49), and ΔEes (r2 = 0.70), all P < 0.05. The inability to sufficiently increase RV O2 supply to meet the increased O2 demand during exercise is principally due to the reduced RV perfusion relative to healthy control values and likely contributes to impaired RV contractile function and thereby to the limited exercise capacity that is commonly observed in patients with PH.NEW & NOTEWORTHY Impaired right ventricular (RV) O2 delivery reserve is associated with reduced RV functional reserve during exercise in a swine model of pulmonary hypertension (PH) induced by pulmonary vein banding. Our data suggest that RV function and exercise capacity might be improved by improving RV O2 delivery.


Enhanced mitophagy in bronchial fibroblasts from severe asthmatic patients.
Sub-epithelial fibrosis is a characteristic feature of airway remodeling in asthma which correlates with disease severity. Current asthma medications are ineffective in treating fibrosis. In this study, we aimed to investigate the mitochondrial phenotype in fibroblasts isolated from airway biopsies of non-asthmatic and severe asthmatic subjects by examining mitophagy as a mechanism contributing to fibroblast persistence and thereby, fibrosis in severe asthma. Bioinformatics analysis of publicly available transcriptomic data was performed to identify the top enriched pathways in asthmatic fibroblasts. Endogenous expression of mitophagy markers in severe asthmatic and non-asthmatic fibroblasts was determined using qRT-PCR, western blot and immunofluorescence. Mitophagy flux was examined by using lysosomal protease inhibitors, E64d and pepstatin A. Mitochondrial membrane potential and metabolic activity were also evaluated using JC-1 assay and MTT assay, respectively. Bioinformatics analysis revealed the enrichment of Pink/Parkin-mediated mitophagy in asthmatic fibroblasts compared to healthy controls. In severe asthmatic fibroblasts, the differential expression of mitophagy genes, PINK1 and PRKN, was accompanied by the accumulation of PINK1, Parkin and other mitophagy proteins at baseline. The further accumulation of endogenous LC3BII, p62 and PINK1 in the presence of E64d and pepstatin A in severe asthmatic fibroblasts reinforced their enhanced mitophagy flux. Significantly reduced mitochondrial membrane potential and metabolic activity were also demonstrated at baseline confirming the impairment in mitochondrial function in severe asthmatic fibroblasts. Interestingly, these fibroblasts displayed neither an apoptotic nor senescent phenotype but a pro-fibrotic phenotype with an adaptive survival mechanism triggered by increased AMPKα phosphorylation and mitochondrial biogenesis. Our results demonstrated a role for mitophagy in the pathogenesis of severe asthma where the enhanced turnover of damaged mitochondria may contribute to fibrosis in severe asthma by promoting the persistence and pro-fibrotic phenotype of fibroblasts.


Age-related impairment of mesenchymal progenitor cell function.
In most mesenchymal tissues a subcompartment of multipotent progenitor cells is responsible for the maintenance and repair of the tissue following trauma. With increasing age, the ability of tissues to repair themselves is diminished, which may be due to reduced functional capacity of the progenitor cells. The purpose of this study was to investigate the effect of aging on rat mesenchymal progenitor cells. Mesenchymal progenitor cells were isolated from Wistar rats aged 3, 7, 12 and 56 weeks. Viability, capacity for differentiation and cellular aging were examined. Cells from the oldest group accumulated raised levels of oxidized proteins and lipids and showed decreased levels of antioxidative enzyme activity. This was reflected in decreased fibroblast colony-forming unit (CFU-f) numbers, increased levels of apoptosis and reduced proliferation and potential for differentiation. These data suggest that the reduced ability to maintain mesenchymal tissue homeostasis in aged mammals is not purely due to a decline in progenitor cells numbers but also to a loss of progenitor functionality due to the accumulation of oxidative damage, which may in turn be a causative factor in a number of age-related pathologies such as arthritis, tendinosis and osteoporosis.


Cotton WRKY1 mediates the plant defense-to-development transition during infection of cotton by Verticillium dahliae by activating JASMONATE ZIM-DOMAIN1 expression.
Plants have evolved an elaborate signaling network to ensure an appropriate level of immune response to meet the differing demands of developmental processes. Previous research has demonstrated that DELLA proteins physically interact with JASMONATE ZIM-DOMAIN1 (JAZ1) and dynamically regulate the interaction of the gibberellin (GA) and jasmonate (JA) signaling pathways. However, whether and how the JAZ1-DELLA regulatory node is regulated at the transcriptional level in plants under normal growth conditions or during pathogen infection is not known. Here, we demonstrate multiple functions of cotton (Gossypium barbadense) GbWRKY1 in the plant defense response and during development. Although GbWRKY1 expression is induced rapidly by methyl jasmonate and infection by Verticillium dahliae, our results show that GbWRKY1 is a negative regulator of the JA-mediated defense response and plant resistance to the pathogens Botrytis cinerea and V. dahliae. Under normal growth conditions, GbWRKY1-overexpressing lines displayed GA-associated phenotypes, including organ elongation and early flowering, coupled with the down-regulation of the putative targets of DELLA. We show that the GA-related phenotypes of GbWRKY1-overexpressing plants depend on the constitutive expression of Gossypium hirsutum GhJAZ1. We also show that GhJAZ1 can be transactivated by GbWRKY1 through TGAC core sequences, and the adjacent sequences of this binding site are essential for binding specificity and affinity to GbWRKY1, as revealed by dual-luciferase reporter assays and electrophoretic mobility shift assays. In summary, our data suggest that GbWRKY1 is a critical regulator mediating the plant defense-to-development transition during V. dahliae infection by activating JAZ1 expression.


Incidental imaging findings from routine chest CT used to identify subjects at high risk of future cardiovascular events.
To investigate the contribution of incidental findings at chest computed tomography (CT) in the detection of subjects at high risk for cardiovascular disease (CVD) by deriving and validating a CT-based prediction rule. This retrospective study was approved by the ethical review board of the primary participating facility, and informed consent was waived. The derivation cohort comprised 10 410 patients who underwent diagnostic chest CT for noncardiovascular indications. During a mean follow-up of 3.7 years (maximum, 7.0 years), 1148 CVD events (cases) were identified. By using a case-cohort approach, CT scans from the cases and from an approximately 10% random sample of the baseline cohort (n = 1366) were graded visually for several cardiovascular findings. Multivariable Cox proportional hazards analysis with backward elimination technique was used to derive the best-fitting parsimonious prediction model. External validation (discrimination, calibration, and risk stratification) was performed in a separate validation cohort (n = 1653). The final model included patient age and sex, CT indication, left anterior descending coronary artery calcifications, mitral valve calcifications, descending aorta calcifications, and cardiac diameter. The model demonstrated good discriminative value, with a C statistic of 0.71 (95% confidence interval: 0.68, 0.74) and a good overall calibration, as assessed in the validation cohort. This imaging-based model allows accurate stratification of individuals into clinically relevant risk categories. Structured reporting of incidental CT findings can mediate accurate stratification of individuals into clinically relevant risk categories and subsequently allow those at higher risk of future CVD events to be distinguished.


[The dynamic detection of cytokins under aplastic anemia and idiopathic thrombocytopenic purpura in the process of impact of hypoxic hypoxia].
The natural hypoxic hypoxia enhances the synthesis processes and decrease the concentration balance in cytokine rets in patients with depression of hematopoiesis. It is established that the patients with aplastic anemia and idiopathic thrombocytopenic purpura, in contrast with patients with acute leucosis, chronic myelolecosis and erythroid myeloma, have quite high values of IL-2, which can be involved in the organization of full-fledgee response to antigen stimuli. The application of hypoxic hypoxia as an additional method in treatment of aplastic anemia and idiopathic thrombocytopenic purpura has no side effects and contraindications.


Physiological links among alternative electron transport pathways that reduce and oxidize plastoquinone in Arabidopsis.
In addition to linear electron transport from water to NADP(+) , alternative electron transport pathways are believed to regulate photosynthesis. In the two routes of photosystem I (PSI) cyclic electron transport, electrons are recycled from the stromal reducing pool to plastoquinone (PQ), generating additional ΔpH (proton gradient across thylakoid membranes). Plastid terminal oxidase (PTOX) accepts electrons from PQ and transfers them to oxygen to produce water. Although both electron transport pathways share the PQ pool, it is unclear whether they interact in vivo. To investigate the physiological link between PSI cyclic electron transport-dependent PQ reduction and PTOX-dependent PQ oxidation, we characterized mutants defective in both functions. Impairment of PSI cyclic electron transport suppressed leaf variegation in the Arabidopsis immutans (im) mutant, which is defective in PTOX. The im variegation was more effectively suppressed in the pgr5 mutant, which is defective in the main pathway of PSI cyclic electron transport, than in the crr2-2 mutant, which is defective in the minor pathway. In contrast to this chloroplast development phenotype, the im defect alleviated the growth phenotype of the crr2-2 pgr5 double mutant. This was accompanied by partial suppression of stromal over-reduction and restricted linear electron transport. We discuss the function of the alternative electron transport pathways in both chloroplast development and photosynthesis in mature leaves.


Adaptations of glutathione antioxidant system to endurance training are tissue and muscle fiber specific.
The effect of endurance training on glutathione (GSH) status and antioxidant enzyme system was investigated in skeletal muscle, heart, and liver of female Sprague-Dawley rats pair fed an isocaloric diet. Ten weeks of treadmill training (25 m/min, 10% grade for 2 h/day, 5 days/wk) increased citrate synthase activity in the deep vastus lateralis (DVL) and soleus muscles by 79 and 39%, respectively (P < 0.01), but not in the heart or liver. In DVL, GSH content was increased 33% (P < 0.05) with training, accompanied by a 64% (P < 0.05) increase in glutamate content but no change in cysteine. Trained rats showed a 62 and 27% higher GSH peroxidase (GPX) and superoxide dismutase (SOD) activity, respectively (P < 0.05), in DVL compared with control rats. In contrast, GSH content and glutathione reductase (GR) activity in soleus declined with training (P < 0.05), whereas activities of GPX and SOD remained unchanged. Training did not alter GSH status in the liver or plasma but significantly decreased the GSH-to glutathione disulfide ratio in the heart. In addition, GR activity in the liver and GSH sulfur-transferase activity in the heart and DVL were significantly lower in the trained vs control rats DVL muscle had threefold higher gamma-glutamyl transpeptidase activity compared with other tissues; however no significant alteration was observed in the activity of gamma-glutamyltranspeptidase or gamma-glutamylcysteine synthetase in the liver, heart, or skeletal muscle. These data indicate that endurance training can cause tissue- and muscle fiber-specific adaptation of antioxidant systems and that GSH homeostasis in extrahepatic tissues may be determined by utilization and uptake of GSH via the gamma-glutamyl cycle.


Studies of regulation of expression of the propionate (prpBCDE) operon provide insights into how Salmonella typhimurium LT2 integrates its 1,2-propanediol and propionate catabolic pathways.
Expression of the prpBCDE operon of Salmonella typhimurium LT2 required (i) the synthesis of propionyl-coenzyme A (CoA) by the PrpE protein or the acetyl-CoA-synthesizing systems of the cell and (ii) the synthesis of 2-methylcitrate from propionyl-CoA and oxaloacetate by the PrpC protein. We propose that either 2-methylcitrate or a derivative of it signals the presence of propionate in the environment. This as yet unidentified signal is thought to serve as a coregulator of the activity of PrpR, the member of the sigma-54 family of transcriptional activators needed for activation of prpBCDE transcription. The CobB protein was also required for expression of the prpBCDE operon, but its role is less well understood. Expression of the prpBCDE operon in cobB mutants was restored to wild-type levels upon induction of the propanediol utilization (pdu) operon by 1,2-propanediol. This effect did not require catabolism of 1,2-propanediol, suggesting that a Pdu protein, not a catabolite of 1,2-propanediol, was responsible for the observed effect. We explain the existence of these redundant functions in terms of metabolic pathway integration. In an environment with 1,2-propanediol as the sole carbon and energy source, expression of the prpBCDE operon is ensured by the Pdu protein that has CobB-like activity. Since synthesis of this Pdu protein depends on the availability of 1,2-propanediol, the cell solves the problem faced in an environment devoid of 1,2-propanediol where propionate is the sole carbon and energy source by having cobB located outside of the pdu operon and its expression independent of 1,2-propanediol. At present, it is unclear how the CobB and Pdu proteins affect prpBCDE expression.


Standardisation of intravenous infusion therapy for paediatrics: big challenges for the little patients.
Worldwide organisations advocate standardising the management of intravenous drugs as an essential strategy to increase safety in paediatric healthcare settings. Intravenous administration is a route associated with some potential complications. Many adverse events are related to the use of intravenous medications, and the great variability in their handling and preparation represents an added risk that jeopardises the safety of children. To standardise the dilutions of intravenous drugs most commonly administered to Spanish hospitalised paediatric and neonatal patients. The process leading to the standardisation of concentrations was undertaken following a two-round modified Delphi procedure. The consensus included the most common drugs administered by continuous or intermittent intravenous infusion to hospitalised and/or critically ill paediatric patients. For paediatric patients, the proposal included a total of 102 drugs (45 continuous infusion and 59 intermittent infusion), with 192 concentrations to be standardised. The final consensus included 101 drugs (99%), of which 44 were continuous infusion and 59 intermittent infusion; 160 concentrations were standardised (72.7%). For neonates, the initial proposal included 80 drugs (38 continuous infusion and 43 intermittent infusion), with 189 concentrations to be standardised. The final consensus included 80 drugs (100%), of which 38 were continuous infusion and 43 were intermittent infusion; 120 concentrations were standardised (49.2%). This proposal showed that standardisation is a feasible approach that can be reached by other healthcare institutions. It can be used in other centres and contribute in the future to unifying paediatric clinical practice.


Absence of cardiolipin in the crd1 null mutant results in decreased mitochondrial membrane potential and reduced mitochondrial function.
Cardiolipin (CL) is a unique phospholipid which is present throughout the eukaryotic kingdom and is localized in mitochondrial membranes. Saccharomyces cerevisiae cells containing a disruption of CRD1, the structural gene encoding CL synthase, have no CL in mitochondrial membranes. To elucidate the physiological role of CL, we compared mitochondrial functions in the crd1Delta mutant and isogenic wild type. The crd1Delta mutant loses viability at elevated temperature, and prolonged culture at 37 degrees C leads to loss of the mitochondrial genome. Mutant membranes have increased phosphatidylglycerol (PG) when grown in a nonfermentable carbon source but have almost no detectable PG in medium containing glucose. In glucose-grown cells, maximum respiratory rate, ATPase and cytochrome oxidase activities, and protein import are deficient in the mutant. The ADP/ATP carrier is defective even during growth in a nonfermentable carbon source. The mitochondrial membrane potential is decreased in mutant cells. The decrease is more pronounced in glucose-grown cells, which lack PG, but is also apparent in membranes containing PG (i.e. in nonfermentable carbon sources). We propose that CL is required for maintaining the mitochondrial membrane potential and that reduced membrane potential in the absence of CL leads to defects in protein import and other mitochondrial functions.


Advanced Design and Manufacture of Mechanoactive Materials Inspired by Skin, Bones, and Skin-on-Bones.
Life is mechanobiological. Natural living materials exhibit remarkable, emergent and smart properties under mechanical loading. Such materials are classified as mechanoactive, in contrast to electroactive polymers and materials that exhibit advanced properties when subjected to electrical stimulation. Cutting edge, multiscale imaging technologies have proven enabling for the elucidation of molecular to meso-scale structure and function of natural mechanoactive materials. Using Microscopy-Aided Design And ManufacturE, (MADAME) this perspective article describes mechanoactive properties of natural materials including skin-on-bones (periosteum) and bone itself. In so doing, it demonstrates the principle to emulate natural smart properties using recursive logic, the basis of many computer algorithms, and to design and manufacture mechanoactive materials and products using advanced manufacturing methods that also incorporate principles of recursive logic. In sum, the MADAME approach translates physically the computer science paradigm of recursion by implementing Jacquard textile methods, which themselves form a historical basis for computing machines, together with additive manufacturing methods including multidimensional printing, stereolithography, laser sintering, etc. These integrated methods provide a foundation and translational pathway for scaled-up manufacture of disruptive mechanoactive materials that will find use in fields as varied as medicine, safety, transport and sports, for internal (implants) and external (wearables) applications.


Drosophila mushroom body mutants are deficient in olfactory learning.
Two Drosophila mutants are described in which the connections between the input to and the output from the mushroom bodies is largely interrupted. In all forms of the flies (larva, imago, male, female) showing the structural defect, olfactory conditioning is impaired. Learning is completely abolished when electroshock is used as reinforcement and partially suppressed in reward learning with sucrose. No influence of the mushroom body defect on the perception of the conditioning stimuli or on spontaneous olfactory behavior is observed. The defect seems not to impair learning of color discrimination tasks or operant learning involving visual cues.


A high-speed single sideband generator using a magnetic tunnel junction spin torque nano-oscillator.
An important property of spin-torque nano-oscillators (STNOs) is their ability to produce a frequency modulated (FM) signal, which is very critical for communication applications. We here demonstrate a novel single sideband (SSB) modulation phenomenon using a magnetic tunnel junction (MTJ)-based STNO, which saves transmission bandwidth and in principle should minimize attenuation for wireless communication. Experimentally, lower single sidebands (LSSBs) have been successfully demonstrated over a wide range of modulation frequency, f m = 150 MHz-1 GHz. The observed LSSBs are determined by the intrinsic properties of the device, which can be modeled well by a nonlinear frequency and amplitude modulation formulation and reproduced in macrospin simulations. Moreover, our macrospin simulation results show that the range of modulation current and modulation frequency for generating SSBs can be controlled by the field-like torque and biasing conditions.


Ultrathin layer solid transformation-enabled-surface enhanced Raman spectroscopy for trace harmful small gaseous molecule detection.
Detection of trace harmful small gaseous molecules (h-SGMs), based on surface enhanced Raman spectroscopy (SERS), has been expected to be a useful strategy but is challenging due to the extremely small Raman cross section (RCS) and weak metal affinity of the h-SGMs. Here, a new strategy, ultrathin layer solid transformation-enabled (ULSTE)-SERS, is proposed. It uses the chemical reaction between the target h-SGM and an ultrathin layer of solid sensing matter coated on a plasmonic metal SERS substrate. This reaction in situ produces a new solid matter with large RCS, which ensures the detection of trace h-SGMs via SERS. The validity of this strategy has been demonstrated by detecting trace H2S gas with an ultrathin CuO layer wrapped around Au nanoparticles. Furthermore, this strategy allows fast and ultrasensitive detection. The detection limit can be down to ppb (even ppt) levels with 10 min preprocessing. Importantly, this strategy has good universality for various other h-SGMs, such as SO2, CS2, CH3SH, and HCl, etc., using appropriate sensing matter. Additionally, the ULSTE-SERS is also suitable for unstable molecules and fast portable detection due to the stable solid layer. This work provides highly efficient SERS-based detection of trace h-SGMs, which is easily applied in practical situations.


Resonance suppression in a TEM cell.
All TEmnp resonances in a TEM cell have been suppressed by cutting the cell walls longitudinally. The TMmnp resonances and propagating TEM mode were not affected. The suppressed resonances have a residual interaction with a biological body inside the cell. Because of this, the modified cell can only be used for bioeffects dosimetry studies at a small number of frequencies above the upper limit for the original cell. Where an accuracy of only +/- 5 dB is required, such as in some EMC studies, the effective bandwidth of the modified cell is twice that of the original cell.


Mapping of quantitative trait loci for the bolting trait in Brassica rapa under vernalizing conditions.
Premature bolting can occur occasionally during spring cultivation of heading Chinese cabbage in East Asia when the plants encounter low temperatures (vernalization), leading to economic loss. Breeding bolting-resistant cultivars is the best choice for solving this problem. We looked for QTLs responsible for varietal differences in the bolting trait in Brassica rapa under environmental conditions that promote vernalization. To achieve this goal, we constructed a linkage map with 107 simple sequence repeats and 54 insertion/deletion markers based on a segregating population of 186 F2 individuals. The resulting map consisted of 10 linkage groups and covered a total length of 947.1 cM, with an average genetic distance of 5.84 cM between adjacent markers. QTL analysis of the bolting trait was performed by two phenotypic evaluations (bolting index and flowering time) based on the scores in an F2 population in the spring of 2010, and scores in F2:3 families in autumn 2010 and spring 2011, respectively. Twenty-six QTLs that controlled bolting were detected, accounting for 2.6 to 31.2% of the phenotypic variance. The detected QTLs with large effects co-localized mainly on linkage groups A02, A06, and A07. These QTLs may provide useful information for marker-assisted selection in a breeding program for late bolting or bolting-resistant cultivars in B. rapa crops.


Identifying communities from multiplex biological networks by randomized optimization of modularity.
The identification of communities, or modules, is a common operation in the analysis of large biological networks. The Disease Module Identification DREAM challenge established a framework to evaluate clustering approaches in a biomedical context, by testing the association of communities with GWAS-derived common trait and disease genes. We implemented here several extensions of the MolTi software that detects communities by optimizing multiplex (and monoplex) network modularity. In particular, MolTi now runs a randomized version of the Louvain algorithm, can consider edge and layer weights, and performs recursive clustering. On simulated networks, the randomization procedure clearly improves the detection of communities. On the DREAM challenge benchmark, the results strongly depend on the selected GWAS dataset and enrichment p -value threshold. However, the randomization procedure, as well as the consideration of weighted edges and layers generally increases the number of trait and disease community detected. The new version of MolTi and the scripts used for the DMI DREAM challenge are available at: https://github.com/gilles-didier/MolTi-DREAM.


The effects of smoking cessation on the risk of chronic obstructive pulmonary disease exacerbations.
Smoking cessation has been demonstrated to reduce the rate of loss of lung function and mortality among patients with mild to moderate chronic obstructive pulmonary disease (COPD). There is a paucity of evidence about the effects of smoking cessation on the risk of COPD exacerbations. We sought to examine whether smoking status and the duration of abstinence from tobacco smoke is associated with a decreased risk of COPD exacerbations. We assessed current smoking status and duration of smoking abstinence by self-report. Our primary outcome was either an inpatient or outpatient COPD exacerbation. We used Cox regression to estimate the risk of COPD exacerbation associated with smoking status and duration of smoking cessation. We performed a cohort study of 23,971 veterans who were current and past smokers and had been seen in one of seven Department of Veterans Affairs (VA) primary care clinics throughout the US. In comparison to current smokers, ex-smokers had a significantly reduced risk of COPD exacerbation after adjusting for age, comorbidity, markers of COPD severity and socio-economic status (adjusted HR 0.78, 95% CI 0.75-0.87). The magnitude of the reduced risk was dependent on the duration of smoking abstinence (adjusted HR: quit < 1 year, 1.04; 95% CI 0.87-1.26; 1-5 years 0.93, 95% CI 0.79-1.08; 5-10 years 0.84, 95% CI 0.70-1.00; > or = 10 years 0.65, 95% CI 0.58-0.74; linear trend <0.001). Smoking cessation is associated with a reduced risk of COPD exacerbations, and the described reduction is dependent upon the duration of abstinence.


Smoking, dietary factors and major age-related eye disorders: an umbrella review of systematic reviews and meta-analyses.
There is accumulating evidence of the associations between age-related eye diseases (AREDs) and smoking or dietary factors. We aimed to provide an umbrella review of the published literature pertaining to smoking or dietary intake as risk factors for major AREDs including cataract, glaucoma, age-related macular degeneration (AMD) and diabetic retinopathy. We searched for pertinent systematic reviews or meta-analyses in PubMed and Web of Science until 16 April 2022. We reperformed the meta-analysis of each association using random effects models. The heterogeneity and 95% prediction interval were calculated. The presence of small-study effect or excess significance bias was also assessed. In total, 64 associations from 25 meta-analyses and 41 associations from 10 qualitative systematic reviews were evaluated. There was convincing (class I) evidence for only one association, namely current smoking and cataract. Two factors had highly suggestive (class II) evidence, namely ever smoking associated with cataract and fish consumption associated with AMD. We also found suggestive (class III) evidence for associations between the dietary intake of omega-3 polyunsaturated fatty acid, lutein, zeaxanthin, vitamin C and the risk of cataract. Smoking as a risk factor for cataract was the most robust association we identified. We also identified several dietary elements associated with AREDs. Large prospective studies are warranted to further examine the associations discussed in this review. CRD42022339082.


[Treatment indications for sarcopenia: a systematic review of exercise intervention effect].
To evaluate exercise intervention efficacy for the prevention and treatment of sarcopenia in the elderly. We conducted a systematic literature search of randomized controlled trials in the Physical Activity Guidelines Advisory Committee Report (before 2007), Pubmed, the Cochrane database, and Igaku Chuo Zasshi (January 2006 to August 2009). Two authors independently extracted relevant data. A total of 951 articles were found by search engines, and 9 studies were finally selected after a review by 2 experts. The content of these studies, especially duration, sets, periods, frequency, and intensity of exercise intervention were extracted and summarized in a results table. Five of 6 articles concluded that high-intensity resistance training significantly increased soft lean tissue and muscle mass. The remaining 3 articles indicated that moderate-intensity resistance training did not affect soft lean tissue or muscle mass. This systematic review suggested that high-intensity resistance training with sufficient periods, frequency, repetitions, and sets is effective to counteract the loss of muscle mass associated with advancing age.


Saffron (Crocus sativus L.) Tea Intake Prevents Learning/Memory Defects and Neurobiochemical Alterations Induced by Aflatoxin B1 Exposure in Adult Mice.
This study aimed to investigate the potential neurotoxic effects of aflatoxin B1 (AFB1) and the preventive effects of saffron. Male Balb-c mice received AFB1 (0.6 mg/kg/day intraperitoneally for 4 days), saffron infusion (90 mg styles/200 mL, ad libitum access for 2 weeks) or saffron infusion plus AFB1 (saffron treatment as previously plus 0.6 mg AFB1/kg/day intraperitoneally for the last 4 days). Control mice were intraperitoneally injected with DMSO:saline (1:1, v/v) during AFB1 treatment. Learning/memory was assessed by passive avoidance task. The activity of acetylcholinesterase [AChE, salt-(SS)/detergent-soluble(DS) isoforms], butyrylcholinesterase (BuChE, SS/DS isoforms), monoamine oxidase (MAO-A, MAO-B), the levels of lipid peroxidation (malondialdehyde, MDA) and reduced glutathione (GSH), were determined in whole brain (minus cerebellum) and cerebellum. We demonstrate for the first time that AFB1 administration impaired the memory of adult mice and decreased significantly whole brain AChE and BuChE activity, cerebellar AChE activity and cerebral GSH content. Moreover, MAO isoforms activity in whole brain, MAO-B activity in cerebellum and MDA levels of both tissues were significantly higher after AFB1 treatment. Pre-treatment with saffron prevented memory decline, activation of MAO-A and MAO-B in whole brain and cerebellum, respectively, and lipid peroxidation triggered by AFB1. Interestingly, the activity of AChE isoforms in whole brain, DS-AChE in cerebellum and GSH levels of both tissues were further significantly decreased in saffron +AFB1-treated mice compared with AFB1 group. Our findings support the neuroprotective efficacy of saffron against AFB1 in adult mice.


Protective effect of zinc on N-methyl-N-nitrosourea and testosterone-induced prostatic intraepithelial neoplasia in the dorsolateral prostate of Sprague Dawley rats.
Previous studies have suggested that zinc exerts anticarcinogenic and antiproliferative effects against prostate cancer both in vitro and in rat ventral prostate. Zinc accumulation diminishes early in the course of prostate malignancy and it inhibits the growth of several carcinoma cells through induction of cell cycle arrest and apoptosis. In this study, we have investigated the influence of zinc on N-methyl-N-nitrosourea (MNU) and testosterone (T)-induced prostatic intraepithelial neoplasia in the dorsolateral prostate of Sprague Dawley (SD) rats. The results indicate that zinc plays an important role in prostate carcinogenesis. Increased tumor incidence was accompanied by a decrease in prostatic acid phosphatase activity, citrate, zinc, glutathione-S-transferase, reduced glutathione, p53, B-cell lymphoma protein (Bcl-2)-associated X protein and caspase-3 levels in MNU + T-treated rats. On the contrary, significantly increased phase I drug metabolizing enzyme activities, lipid peroxide, hydrogen peroxide, proliferating cell nuclear antigen, Bcl-2 and Bcl-X(L) protein levels were observed in the dorsolateral prostate of MNU + T-treated rats. Simultaneous zinc supplementation significantly reversed these effects in MNU + T-treated rats. Signs of dysplasia, a characteristic of prostatic intraepithelial neoplasia, were evident in the dorsolateral prostatic tissue sections by MNU + T administration. However, zinc supplementation has reversed these effects in the dorsolateral prostatic histoarchitecture. These results suggest that zinc may act as an essential trace element against MNU and testosterone-induced prostatic preneoplastic progression in SD rats.


A B Cell-Driven Autoimmune Pathway Leading to Pathological Hallmarks of Progressive Multiple Sclerosis in the Marmoset Experimental Autoimmune Encephalomyelitis Model.
The absence of pathological hallmarks of progressive multiple sclerosis (MS) in commonly used rodent models of experimental autoimmune encephalomyelitis (EAE) hinders the development of adequate treatments for progressive disease. Work reviewed here shows that such hallmarks are present in the EAE model in marmoset monkeys (Callithrix jacchus). The minimal requirement for induction of progressive MS pathology is immunization with a synthetic peptide representing residues 34-56 from human myelin oligodendrocyte glycoprotein (MOG) formulated with a mineral oil [incomplete Freund's adjuvant (IFA)]. Pathological aspects include demyelination of cortical gray matter with microglia activation, oxidative stress, and redistribution of iron. When the peptide is formulated in complete Freund's adjuvant, which contains mycobacteria that relay strong activation signals to myeloid cells, oxidative damage pathways are strongly boosted leading to more intensive pathology. The proven absence of immune potentiating danger signals in the MOG34-56/IFA formulation implies that a narrow population of antigen-experienced T cells present in the monkey's immune repertoire is activated. This novel pathway involves the interplay of lymphocryptovirus-infected B cells with MHC class Ib/Caja-E restricted CD8+ CD56+ cytotoxic T lymphocytes.


Comparison of glatiramer acetate (Copaxone) and interferon beta-1b (Betaferon) in multiple sclerosis patients: an open-label 2-year follow-up.
To compare the clinical efficacy, as expressed by relapse rate and disability accumulation, and safety profile of glatiramer acetate (Copaxone; COP-1) and Interferon beta-1b (Betaferon; IFN beta - 1b) administered to multiple sclerosis patients during a 2-year follow-up on an open-label parallel design, as compared to their clinical condition in the 2-year period prior to treatment. Copaxone and IFN beta - 1b have been recently introduced for the treatment of relapsing forms of MS. Both medications have been proven to have a relatively safe profile and are used extensively world-wide. 58 consecutive patients with relapsing forms of MS were enrolled from the MS out-patient clinic, during three months. After being informed in detail of the two approved treatment options existing at the time in Israel, the patients chose by themselves to receive either: (a) Copaxone 20 mg subcutaneously (sc) daily (Copaxone dly, 20 patients), or (b) Copaxone 20 mg sc alternate-day (Copaxone alt, 18 patients) or (c) IFN beta-1b 8 MIU sc in alternate day (20 patients). Mean relapse rate/year and mean EDSS/year were calculated for each group of patients during the 2 years prior to the onset of treatment, and during the year prior to the onset of treatment. Statistical significance was observed in the relapse rate in the year prior to the onset of treatment between the IFN beta -1b group and the two Copaxone groups (p = 0.05). This statistical difference has no effect on the overall data of the 2 years prior to starting the treatment and on the results. No statistical significance was observed in the total number of relapses, and on the 2-year relapse rate, prior to the onset of treatment. Mean relapse rate/year and mean EDSS/year were calculated for each group during the first and second year of treatment. Wilcoxon analysis for clinical data and chi-square for adverse events were applied. The three groups were statistically comparable concerning mean relapse/year in the 2 years before the trial started and no statistical significance was observed among the three groups. A statistically significant reduction in the mean relapse rate in the 2 years after onset of treatment was observed in the three group of patients: Copaxone daily (dly) 1.1 +/- 0.6 (p = 0.0001); Copaxone alternate (alt) 0.9 +/- 0.6 (p = 0.0004) and IFN beta -1b 1.2 +/- 0.7 (p = 0.0001). Disability as expressed by EDSS score prior to the onset of treatment and after 2 years of treatment showed deterioration in the three groups although more significant in the Copaxone groups: Copaxone dly 3.3 +/- 1.4 to 3.8 +/- 1.6 (p = 0.007); Copaxone alt 2.4 +/- 1.1 to 2.8 +/- 1.3 (p=0.04); IFN beta - 1b 3.1 +/- 1.3 to 3.3 +/- 2.0 (N.S.). The most common adverse events reported were: (1) flu-like symptoms 7 pts (35%) in the IFN beta -1b group; 10 pts (26%) of the two Copaxone groups; (2) increased spasticity of lower limbs 3 pts (15%), only in the IFN beta -1b group; (3) site injection reaction (SIR): 16 SIR (80%) in the IFN beta -1b group; 12 SIR (67%) in the Copaxone alt group; 14 SIR (70%) in the Copaxone dly group; and (4) systemic reaction 3 pts (15%) in the IFN beta -1b group; 4 pts (22%) in the Copaxone alt group; 6 pts (30%) in the Copaxone dly group. Premature termination occurred in five patients treated with Copaxone (3 in the alternate group and 2 in the daily group). The present study, despite the limitations of an open-label study, shows that Copaxone dly, Copaxone alt and IFN beta -1b treatment seem to be equally effective for the control of exacerbations in MS. The adverse event profile, as reported by the patients, was also similar. However, the adverse events profile registered indicated that Copaxone is somewhat less detrimental, whereas disability as measured by EDSS accumulation showed that the interferon beta - 1b patients demonstrated a slower progression of the disability.


The application of k-space in pulse echo ultrasound.
K-space is a frequency domain description of imaging systems and targets that can be used to gain insight into image formation. Although originally proposed in ultrasound for the analysis of experiments involving anisotropic scattering and for the design of acoustic tomography systems, it is particularly useful for the analysis of pulse echo ultrasonic imaging systems. This paper presents analytical and conceptual techniques for estimating the k-space representation of pulse echo imaging systems with arbitrary transmit and receive array geometries and apodizations. Simple graphical methods of estimating the first and second order statistics of speckle are presented. Examples are presented that utilize k-space to gain insight regarding the performance of spatial and frequency compounding and to describe the impact of synthetic receive aperture geometry on beamforming and speckle statistics. The Van Cittert-Zernike theorem is presented in k-space, and techniques for improving echo coherence are discussed.


Effects of lithium administration on plasma catecholamines.
The mode of action of lithium in the treatment of bipolar affective disorder is unknown. Among its actions are effects on release and response to catecholamines in experimental animals. We have therefore examined its effect on peripheral catecholamines in man. After 3 weeks of administration of therapeutic doses of lithium carbonate, healthy volunteers showed a differential response of catecholamines to insulin stimulation. Although the response of plasma norepinephrine remained unchanged, epinephrine response was dramatically reduced. These findings provide additional evidence for the separate neural regulation of the adrenal medulla and sympathetic nerve endings. Further studies are necessary to determine whether these effects on peripheral epinephrine are paralleled by changes in central epinephrine.