Review of renal anastomosing hemangioma with focus on clinical and pathological aspects.
Renal anastomosing hemangiomas (RAH) has been recently proposed as a new entity. In this article, we summarize the clinicopathologic features of this tumor. RAH usually develops on a background of end-stage renal disease. Macroscopically, tumors are well-defined and their cut surface shows mahogany brown spongy tissue with epicenter in the renal medulla. Tumors are usually small, but larger lesions are reported. On microscopic examination, the tumor consists of sinusoid-like vascular channels lined by cuboidal endothelial cells with occasional hobnail-like appearance of endothelial cells closely mimicking splenic sinusoids. Eosinophilic hyaline globules may be present in the cytoplasm of neoplastic endothelial cells. Extramedullary hematopoiesis containing erythroid precursor and megakaryocytes may be present in the vascular lumens. Immunohistochemically, endothelial cells are positive for CD31 and CD34, but negative for D2-40, GLUT-1 and HHV8. The surrounding stroma around endothelial cells demonstrates positivity for  smooth muscle action. To date, there are no studies on molecular genetic aspects of RAH. This tumor is indolent based on site and size of the lesion, partial or nephrectomy is sufficient as a therapeutic modality.


Interleukin-1 receptor antagonist reverses stroke-associated peripheral immune suppression.
Infections are common following stroke and adversely affect outcome. Cellular immune suppression associated with acute stroke may increase susceptibility to infection. Cytokines are important contributors to both stroke pathology and the response to infection. Since interleukin (IL)-1 blockade is a candidate treatment for cerebral ischemia, we examined whether administration of interleukin-1 receptor antagonist (IL-1Ra) to patients with acute stroke affected innate cellular immune responses in a phase II placebo-controlled trial. Venous blood samples were taken prior to treatment initiation, at 24h and 5 to 7d. Blood was also drawn from stroke-free controls. Lipopolysaccharide (LPS) stimulation of whole-blood cultures assessed the potential of leukocytes to produce cytokines. Induction of tumor necrosis factor (TNF)-α, IL-1β, IL-6, IL-8 and IL-10 by LPS was significantly reduced in patients at admission, compared to controls. At 24h, cytokine induction remained suppressed in the placebo group. In contrast, for patients treated with IL-1Ra, induction of TNF-α, IL-6 and IL-10 was similar to controls and IL-1β induction was significantly greater than in the placebo group. At 5 to 7d, TNF-α and IL-1β induction remained suppressed only in the placebo group (p<0.05). Plasma cortisol concentrations, elevated at admission in patients compared to controls, were substantially reduced at 24h in the patients receiving IL-1Ra (p<0.05) and inversely correlated (p<0.001) with either TNF-α (r=-0.71) or IL-1β induction (r=-0.67) at admission. Treatment with IL-1Ra reverses peripheral innate immune suppression in the acute phase of stroke, which is associated with attenuated cortisol production. The mechanisms underlying these observations, including the potential impact of IL-1Ra on stroke severity and the clinical significance of immune suppression, require further evaluation in larger studies.


Activation of 1-alkyl-2-lysoglycero-3-phosphocholine. Acetyl-CoA transferase during phagocytosis in human polymorphonuclear leukocytes.
1-Alkyl-2-acetyl-sn-glycero-3-phosphocholine can be synthesized either by acetylation of 1-alkyl-2-lyso-sn-glycero-3-phosphocholine or by transcholination of 1-alkyl-2-acetyl-sn-glycerol. Addition of opsonized zymosan to isolated human polymorphonuclear leukocytes induces a fast, transient, up to 10-fold activation of the acetyltransferase reaction without affecting the cholinephosphotransferase pathway. Based on the following results, we propose that the generation of 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine by human polymorphonuclear leukocytes during phagocytosis is mediated by a stimulation of the acetyltransferase reaction: 1) the time course of acetyltransferase activation agrees with it being the enzyme responsible for the synthesis of 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine; and 2) a close correlation exists (r = 0.91) between the generation of 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine in response to various doses of zymosan and the magnitude of acetyltransferase activation.


Aridity is expressed in river topography globally.
It has long been suggested that climate shapes land surface topography through interactions between rainfall, runoff and erosion in drainage basins1-4. The longitudinal profile of a river (elevation versus distance downstream) is a key morphological attribute that reflects the history of drainage basin evolution, so its form should be diagnostic of the regional expression of climate and its interaction with the land surface5-9. However, both detecting climatic signatures in longitudinal profiles and deciphering the climatic mechanisms of their development have been challenging, owing to the lack of relevant global data and to the variable effects of tectonics, lithology, land surface properties and human activities10,11. Here we present a global dataset of 333,502 river longitudinal profiles, and use it to explore differences in overall profile shape (concavity) across climate zones. We show that river profiles are systematically straighter with increasing aridity. Through simple numerical modelling, we demonstrate that these global patterns in longitudinal profile shape can be explained by hydrological controls that reflect rainfall-runoff regimes in different climate zones. The most important of these is the downstream rate of change in streamflow, independent of the area of the drainage basin. Our results illustrate that river topography expresses a signature of aridity, suggesting that climate is a first-order control on the evolution of the drainage basin.


Sexual variation in heritability and genetic correlations of morphological traits in house sparrow (Passer domesticus).
Estimates of genetic components are important for our understanding of how individual characteristics are transferred between generations. We show that the level of heritability varies between 0.12 and 0.68 in six morphological traits in house sparrows (Passer domesticus L.) in northern Norway. Positive and negative genetic correlations were present among traits, suggesting evolutionary constraints on the evolution of some of these characters. A sexual difference in the amount of heritable genetic variation was found in tarsus length, wing length, bill depth and body condition index, with generally higher heritability in females. In addition, the structure of the genetic variance-covariance matrix for the traits differed between the sexes. Genetic correlations between males and females for the morphological traits were however large and not significantly different from one, indicating that sex-specific responses to selection will be influenced by intersexual differences in selection differentials. Despite this, some traits had heritability above 0.1 in females, even after conditioning on the additive genetic covariance between sexes and the additive genetic variances in males. Moreover, a meta-analysis indicated that higher heritability in females than in males may be common in birds. Thus, this indicates sexual differences in the genetic architecture of birds. Consequently, as in house sparrows, the evolutionary responses to selection will often be larger in females than males. Hence, our results suggest that sex-specific additive genetic variances and covariances, although ignored in most studies, should be included when making predictions of evolutionary changes from standard quantitative genetic models.


[Preparation of dihydrocarminomycin and a comparison of its antitumor activity with the activity of carminomycin].
A dihydro derivative of karminomycin was prepared using chemical reduction with potassium boron hydride. When dihydrokarminomycin was administered intravenously to healthy albino mice in a single dose it practically showed the same toxicity as karminomycin. However, unlike the latter dihydrokarminomycin induced the death of the animals at later periods of time. Studies on mice with transplantable tumours showed high antitumor activity of dihydrokarminomycin against lymphosarcoma L10-1, sarcoma 180, Garding-Passy melanoma, lymphoid leukosis L-1210 and lymphocytal leukosis P-388. In treatment of the mice with leukosis L-1210 and Garding-Passy melanoma dihydrokapminomycin was much inferior by its efficiency than karminomycin.


[Continuous and intermittent exercise: effects of training and detraining on body fat in obese rats].
Exercise training is often recommended in prevention and treatment of obesity. The present study was designed to compare the effects of intermittent and continuous exercise on weight loss and carcass composition in obese rats. Obese male Wistar rats (monosodium glutamate [MSG] administration, 4 mg/g of body weight every other day from birth to 14 days old) were used. After drug administration, the rats were separated into three groups: MSG-SED (sedentary), MSG-CONT (continuous, swimming, 45 min/day, 5 days/week, with and overload of 5% body weight for 12 weeks) and MSG-INT (intermittent, 15 s swimming intermitted by 15 s rest, during 45 min, 5 days/week, with and overload of 15% body weight for 12 weeks). Rats of the same age and strain, administered with saline were used as control (SAL), and subdivided into three groups: SAL-SED, SAL-CONT and SAL-INT. The animals were evaluated at the 10 weeks of training and 8 weeks of its interruption. MSG rats showed higher carcass fat as well as weight and cell size in epididymal adipose tissue than SAL rats, indicting the efficacy of the drug in producing obesity. Intermittent training protocol led to a reduction in blood lactate accumulation during acute exercise and both protocols reduced body weight gain during the experiment in MSG rats. After 8 weeks of training interruption no differences were observed among groups in the examined parameters. Only intermittent exercise training improved aerobic fitness but both protocols were similarly efficient in determining weight loss. However, the effects were transitory, since they disappeared after detraining.


Clinical and molecular correlates in fragile X premutation females.
The prevalence of the fragile X premutation (55-200 CGG repeats) among the general population is relatively high, but there remains a lack of clear understanding of the links between molecular biomarkers and clinical outcomes. In this study we investigated the correlations between molecular measures (CGG repeat size, FMR1 mRNA, FMRP expression levels, and methylation status at the promoter region and in FREE2 site) and clinical phenotypes (anxiety, obsessive compulsive symptoms, depression and executive function deficits) in 36 adult premutation female carriers and compared to 24 normal control subjects. Premutation carriers reported higher levels of obsessive compulsive symptoms, depression, and anxiety, but demonstrated no significant deficits in global cognitive functions or executive function compared to the control group. Increased age in carriers was significantly associated with increased anxiety levels. As expected, FMR1 mRNA expression was significantly correlated with CGG repeat number. However, no significant correlations were observed between molecular (including epigenetic) measures and clinical phenotypes in this sample. Our study, albeit limited by the sample size, establishes the complexity of the mechanisms that link the FMR1 locus to the clinical phenotypes commonly observed in female carriers suggesting that other factors, including environment or additional genetic changes, may have an impact on the clinical phenotypes. However, it continues to emphasize the need for assessment and treatment of psychiatric problems in female premutation carriers.


Comparison of caspase activation and subcellular localization in HL-60 and K562 cells undergoing etoposide-induced apoptosis.
Previous studies have shown that K562 chronic myelogenous leukemia cells are resistant to induction of apoptosis by a variety of agents, including the topoisomerase II (topo II) poison etoposide, when examined 4 to 24 hours after treatment with an initiating stimulus. In the present study, the responses of K562 cells and apoptosis-proficient HL-60 acute myelomonocytic leukemia cells to etoposide were compared, with particular emphasis on determining the long-term fate of the cells. When cells were treated with varying concentrations of etoposide for 1 hour and subsequently plated in soft agar, the two cell lines displayed similar sensitivities, with a 90% reduction in colony formation at 5 to 10 mu mol/L etoposide. After treatment with 17 mu mol/L etoposide for 1 hour, cleavage of the caspase substrate poly(ADP-ribose) polymerase (PARP), DNA fragmentation, and apoptotic morphological changes were evident in HL-60 cells in less than 6 hours. After the same treatment, K562 cells arrested in G2 phase of the cell cycle but otherwise appeared normal for 3 to 4 days before developing similar apoptotic changes. When the etoposide dose was increased to 68 mu mol/L, apoptotic changes were evident in HL-60 cells after 2 to 3 hours, whereas the same changes were observed in K562 cells after 24 to 48 hours. This delay in the development of apoptotic changes in K562 cells was accompanied by delayed release of cytochrome c to the cytosol and delayed appearance of peptidase activity that cleaved the fluorogenic substrates Asp-Glu-Val-Asp-aminotrifluoromethylcoumarin (DEVD-AFC) and Val-Glu-Ile-Asp-aminomethylcoumarin (VEID-AMC) as well as an altered spectrum of active caspases that were affinity labeled with N-(Nalpha-benzyloxycarbonylglutamyl-Nepsilon-biotin yllysyl) aspartic acid [(2,6-dimethylbenzoyl)oxy]methyl ketone [z-EK(bio)D-aomk]. On the other hand, the activation of caspase-3 under cell-free conditions occurred with indistinguishable kinetics in cytosol prepared from the two cell lines. Collectively, these results suggest that a delay in the signaling cascade upstream of cytochrome c release and caspase activation leads to a long latent period before the active phase of apoptosis is initiated in etoposide-treated K562 cells. Once the active phase of apoptosis is initiated, the spectrum and subcellular distribution of active caspase species differ between HL-60 and K562 cells, but a similar proportion of cells are ultimately killed in both cell lines.


Association between phosphatidylinositol 3-kinase regulatory subunit p85alpha Met326Ile genetic polymorphism and colon cancer risk.
The phosphatidylinositol 3-kinase signaling pathway is frequently activated in cancer. Emerging evidence supports the p85alpha regulatory subunit gene, PIK3R1, as a novel oncogene. We examined the association of a functional missense polymorphism (Met326Ile) of PIK3R1 with colon cancer risk in a population-based case-control study of 421 incident cases and 483 controls. In our base unconditional logistic regression model controlling for age, gender, and race, we observed a 47% increase in risk among those carrying one or two copies of the 326Ile variant (P = 0.01). Further adjustment for family history of colorectal cancer, body mass index, nonsteroidal anti-inflammatory drugs, smoking, alcohol consumption, and physical activity strengthened the association [odds ratio (OR), 1.73; 95% confidence interval (CI), 1.24-2.42, P = 0.001]. The association was more pronounced among those older than 64 years (OR, 2.10; 95% CI, 1.19-3.70, P = 0.01). Evaluation of the genotypes assuming an additive mode of inheritance showed a significant trend for gene-dose response, where compared with Met/Met, the OR estimates for Ile/Met and Ile/Ile were 1.68 (95% CI, 1.19-2.37) and 2.27 (95% CI, 0.98-5.29), respectively (P for trend = 0.001). This study is the first to describe a significant association between a germ line functional variant in PIK3R1 and cancer, providing new evidence supporting a role for PIK3R1 in the development of colon cancer.


Examining therapy duration in adults with induced laryngeal obstruction (ILO).
This study examined the number of therapy sessions required to sufficiently improve (exercise) induced laryngeal obstruction (EILO/ILO) symptoms for discharge. Factors predicting therapy duration were examined as was the likelihood of patients returning for additional therapy sessions following initial discharge. Retrospective observational cohort design. Data for 350 patients were gathered from the University of Wisconsin-Madison Voice and Swallow Clinics Outcome Database. Patients (>18 years of age) diagnosed with EILO/ILO received therapy from a Speech-Language Pathologist (SLP) and were successfully discharged. EILO/ILO treatment details, symptoms, triggers, medical comorbidities, and patient demographics were collected from initial evaluations and subsequent course of therapy. Patients required an average of 3.59 (SD = 3.7) therapy sessions prior to discharge. A comorbid behavioral health diagnosis (p = .026), higher Vocal Handicap Index Score (p = .009) and reduced physical activity due to EILO/ILO symptoms (p = .032) were associated with increased therapy duration. Patients with ILO or EILO with secondary environmental triggers required significantly more sessions than those with exercise-induced symptoms (p < .01). Eight percent of patients returned for additional sessions following discharge. Patients returning for additional sessions all came from affluent neighborhoods as measured by the Area Deprivation Index (ADI). Patients with EILO/ILO required an average of 3.59 therapy sessions prior to discharge. As such, 4 sessions is a reasonable estimate for clinicians to provide patients. Six sessions may be a more conservative estimate for patients who present with a behavioral health diagnosis, a voice complaint, or reduced physical activity from EILO/ILO symptoms.


Biological Regeneration of Articular Cartilage in an Early Stage of Compartmentalized Osteoarthritis: 12-Month Results.
Biological regeneration in an early stage of osteoarthritis (OA) is an important clinical challenge. An early-stage compartmentalized OA model was used to evaluate different biological regeneration techniques. Biological regeneration in an early stage of compartmentalized OA is possible. Controlled laboratory study. A 7-mm cartilage defect was surgically created in 24 sheep. After 3 months, by which time early OA had set in, the sheep were randomized into 4 different treatment groups and operated for the second time. One group (CONTROL) served as a long-term follow-up group for the further development of OA. The other 3 groups (regeneration groups) each underwent a different regeneration procedure after abrasion of the subchondral bone (defect size: 20 × 10 mm with a depth of 2.5 mm): spongialization alone (SPONGIO), spongialization followed by implantation of an unseeded hyaluronan matrix (MATRIX), or spongialization followed by implantation of a hyaluronan matrix seeded with autologous chondrocytes (MACT). Then, 12 months after the second operative procedure, the animals were euthanized and the defects subjected to macroscopic and histological grading. Historical 4-month data were compared with the 12-month results. After 12 months of follow-up, advanced cartilage degeneration was observed in the CONTROL group. On the other hand, all regeneration groups improved significantly compared with the 4-month results using the Mankin score. Cartilage quality in the MACT group was significantly better than in the MATRIX group, as determined by the Mankin and the O'Driscoll scores. There are no existing clinical options for preventing early OA from progressing to a severe disease. This study provides important information on how a surgical intervention can forestall the development of OA. OA of the knee is very common. Total joint replacement is not an acceptable option for active patients. Biological regeneration in OA is successful for focal cartilage defects; however, a long-term follow-up for biological regeneration in OA is missing. It is essential to have long-term results for a regenerative procedure involving cartilage, which is a tissue with a very slow turnover.


⁵¹V NMR Crystallography of Vanadium Chloroperoxidase and Its Directed Evolution P395D/L241V/T343A Mutant: Protonation Environments of the Active Site.
Vanadium-dependent haloperoxidases (VHPOs) perform two-electron oxidation of halides using hydrogen peroxide. Their mechanism, including the factors determining the substrate specificity and the pH-dependence of the catalytic rates, is poorly understood. The vanadate cofactor in the active site of VHPOs contains "spectroscopically silent" V(V), which does not change oxidation state during the reaction. We employed an NMR crystallography approach based on (51)V magic angle spinning NMR spectroscopy and Density Functional Theory, to gain insights into the structure and coordination environment of the cofactor in the resting state of vanadium-dependent chloroperoxidases (VCPO). The cofactor environments in the wild-type VCPO and its P395D/L241V/T343A mutant exhibiting 5-100-fold improved catalytic activity are examined at various pH values. Optimal sensitivity attained due to the fast MAS probe technologies enabled the assignment of the location and number of protons on the vanadate as a function of pH. The vanadate cofactor changes its protonation from quadruply protonated at pH 6.3 to triply protonated at pH 7.3 to doubly protonated at pH 8.3. In contrast, in the mutant, the vanadate protonation is the same at pH 5.0 and 8.3, and the cofactor is doubly protonated. This methodology to identify the distinct protonation environments of the cofactor, which are also pH-dependent, could help explain the different reactivities of the wild-type and mutant VCPO and their pH-dependence. This study demonstrates that (51)V-based NMR crystallography can be used to derive the detailed coordination environments of vanadium centers in large biological molecules.


Comparison of pain response after subcutaneous injection of two maropitant formulations to beagle dogs.
The antiemetic maropitant, with metacresol as preservative (Cerenia, Zoetis), has been associated with pain after subcutaneous injection in dogs and cats. Recently, a generic formulation containing benzyl alcohol was authorised (Prevomax, Le Vet). Benzyl alcohol is reported to have local anaesthetic properties and reduce injection pain. This study compared local pain after subcutaneous injection of the two maropitant formulations, administered at approximately 4°C and 25°C, to dogs. Thirty-two healthy beagle dogs were enrolled into a blinded, randomised, cross-over study. Dogs received subcutaneous injections of maropitant injection containing metacresol as preservative and maropitant injection containing benzyl alcohol as preservative, both at approximately 4°C and 25°C, with at least three days in between treatments. Injection pain was evaluated by two blinded observers using a visual analogue scale immediately after injection and a simple descriptive scale at two minutes after injection. In healthy beagle dogs, subcutaneous injection of maropitant with benzyl alcohol is significantly less painful than injection of maropitant with metacresol.


Cord blood and breast milk iron status in maternal anemia.
The purpose of this work was to assess the effect of severe maternal iron-deficiency anemia and nutritional status on cord blood and breast milk iron status. We conducted a prospective observational study over a 6-month period in a teaching hospital in central India. The study population consisted of 55 anemic (hemoglobin: < 110 g/L) and 20 healthy nonanemic (hemoglobin: > or = 110 g/L) pregnant women who delivered singleton live births at term gestation. We measured hemoglobin, iron, and ferritin levels in paired maternal and cord blood and iron levels in early (day 3 +/- 1) and late (day 15 +/- 3) transitional milk. Maternal anthropometry, including weight, height, midarm circumference, triceps skinfold thickness, and placental weight, were recorded. The main outcome measure of the study was to find out the relationship of maternal hemoglobin, iron, ferritin, and anthropometry with hemoglobin, iron, and ferritin in cord blood and iron levels in breast milk. Concentrations of hemoglobin, iron, and ferritin were significantly lower in the cord blood of anemic mothers and showed linear relationships with maternal hemoglobin and ferritin levels. Breast milk iron content was significantly reduced in severely anemic mothers but not in those with mild-to-moderate anemia. Breast milk iron level correlated with maternal hemoglobin and iron levels but not with ferritin levels. Maternal anthropometry had significant correlations with indices of iron nutriture in maternal and cord blood but showed no relationship with breast milk iron content. Placental weight was comparable between anemic and nonanemic mothers. Maternal anemia, particularly the severe type, adversely affects cord blood and breast milk iron status. Maternal nutritional status exerts a significant influence on fetal iron status but has little influence on breast milk iron content.


Mechanism of p21-activated kinase 6-mediated inhibition of androgen receptor signaling.
PAK6 was first identified as an androgen receptor (AR)-interacting protein able to inhibit AR-mediated transcriptional responses. PAK6 is a serine/threonine kinase belonging to the p21-activated kinase (PAK) family implicated in actin reorganization and cell motility, gene transcription, apoptosis, and cell transformation. We investigated the biochemical basis for inhibition of AR signaling by PAK6. We compared the kinase activity of PAK6 with two other well characterized members of the PAK family, PAK1 and PAK4. Like PAK4, PAK6 possesses a constitutive basal kinase activity that, unlike PAK1, is not modulated by the binding of active Rac or Cdc42 GTPases. In order to test the involvement of PAK6 kinase activity in suppression of AR-mediated transcription, we generated kinase-dead (K436A) and kinase-active (S531N) mutants of PAK6. We show that PAK6 kinase activity is required for effective PAK6-induced repression of AR signaling. Suppression does not depend upon GTPase binding to PAK6 and is not mimicked by the closely related PAK1 and PAK4 isoforms. Kinase-dependent inhibition by PAK6 extended to the enhanced AR-mediated transcription seen in the presence of coactivating molecules and to the action of AR coinhibitors. Active PAK6 inhibited nuclear translocation of the stimulated AR, suggesting a possible mechanism for inhibition of AR responsiveness. Finally, we observe that autophosphorylated, active PAK6 protein is differently expressed among prostate cancer cell lines. Modulation of PAK6 activity may be responsible for regulation of AR signaling in various forms of prostate cancer.


Next generation maternal health: external shocks and health-system innovations.
In this Series we document the substantial progress in the reduction of maternal mortality and discuss the current state of science in reducing maternal mortality. However, maternal health is also powerfully influenced by the structures and resources of societies, communities, and health systems. We discuss the shocks from outside of the field of maternal health that will influence maternal survival including economic growth in low-income and middle-income countries, urbanisation, and health crises due to disease outbreaks, extreme weather, and conflict. Policy and technological innovations, such as universal health coverage, behavioural economics, mobile health, and the data revolution, are changing health systems and ushering in new approaches to affect the health of mothers. Research and policy will need to reflect the changing maternal health landscape.


Molecular virology of Epstein-Barr virus.
Epstein-Barr virus (EBV) interacts with its host in three distinct ways in a highly regulated fashion: (i) EBV infects human B lymphocytes and induces proliferation of the infected cells, (ii) it enters into a latent phase in vivo that follows the proliferative phase, and (iii) it can be reactivated giving rise to the production of infectious progeny for reinfection of cells of the same type or transmission of the virus to another individual. In healthy people, these processes take place simultaneously in different anatomical and functional compartments and are linked to each other in a highly dynamic steady-state equilibrium. The development of a genetic system has paved the way for the dissection of those processes at a molecular level that can be studied in vitro, i.e. B-cell immortalization and the lytic cycle leading to production of infectious progeny. Polymerase chain reaction analyses coupled to fluorescent-activated cell sorting has on the other hand allowed a descriptive analysis of the virus-host interaction in peripheral blood cells as well as in tonsillar B cells in vivo. This paper is aimed at compiling our present knowledge on the process of B-cell immortalization in vitro as well as in vivo latency, and attempts to integrate this knowledge into the framework of the viral life cycle in vivo.


Erythromycin or Clindamycin - is it Still an Empirical Therapy against Streptococcus agalactiae in Patients Allergic to Penicillin?
Retrospective analysis of Streptococcus agalactiae antibiotic susceptibility isolated in 2010-2013 was performed. Penicillin was still the first-line antibiotic. Due to the high percentage of strains resistant to erythromycin and clindamycin empirical treatment with these antibiotics may not be effective. Lower resistance rate to erythromycin and clindamycin among strains isolated from infected pregnant women and newborns were observed than among strains isolated from samples from patients hospitalized in other departments (29% and 47% v. 46% and 63%). The increasing resistance rate might give a rise to a new epidemiological situation.


Endo Mini-Retract(TM) laparoscopic retractor with a novel short-cut Nelaton catheter for dividing the vasculature in laparoscopic liver resection.
Laparoscopic hepatectomy has become a widespread procedure in Japan, now that the national medical insurance system covers partial resection and lateral segmentectomy. An important procedure during laparoscopic hepatectomy is the division of the vascular pedicles. In this paper we report a practical and useful method for vascular division with an Endo Mini-Retract(TM) retractor and a shortened Nelaton catheter (2.5-mm-diameter) developed in our department.


Anti-GM-CSF otilimab versus tofacitinib or placebo in patients with active rheumatoid arthritis and an inadequate response to conventional or biologic DMARDs: two phase 3 randomised trials (contRAst 1 and contRAst 2).
To investigate the efficacy and safety of otilimab, an antigranulocyte-macrophage colony-stimulating factor antibody, in patients with active rheumatoid arthritis. Two phase 3, double-blind randomised controlled trials including patients with inadequate responses to methotrexate (contRAst 1) or conventional synthetic/biologic disease-modifying antirheumatic drugs (cs/bDMARDs; contRAst 2). Patients received background csDMARDs. Through a testing hierarchy, subcutaneous otilimab (90/150 mg once weekly) was compared with placebo for week 12 endpoints (after which, patients receiving placebo switched to active interventions) or oral tofacitinib (5 mg two times per day) for week 24 endpoints. proportion of patients achieving an American College of Rheumatology response ≥20% (ACR20) at week 12. The intention-to-treat populations comprised 1537 (contRAst 1) and 1625 (contRAst 2) patients. proportions of ACR20 responders were statistically significantly greater with otilimab 90 mg and 150 mg vs placebo in contRAst 1 (54.7% (p=0.0023) and 50.9% (p=0.0362) vs 41.7%) and contRAst 2 (54.9% (p<0.0001) and 54.5% (p<0.0001) vs 32.5%). Secondary endpoints: in both trials, compared with placebo, otilimab increased the proportion of Clinical Disease Activity Index (CDAI) low disease activity (LDA) responders (not significant for otilimab 150 mg in contRAst 1), and reduced Health Assessment Questionnaire-Disability Index (HAQ-DI) scores. Benefits with tofacitinib were consistently greater than with otilimab across multiple endpoints. Safety outcomes were similar across treatment groups. Although otilimab demonstrated superiority to placebo in ACR20, CDAI LDA and HAQ-DI, improved symptoms, and had an acceptable safety profile, it was inferior to tofacitinib. NCT03980483, NCT03970837.


Operating room nurses' perceptions of the effects of physician-perpetrated abuse.
Operating room (OR) nurses experience abuse perpetrated by physicians; however, little research has been conducted to examine nurses' perceptions of the effects of such abuse. The aim of this research was to understand participants' perceptions of physician-perpetrated abuse on their health and ability to provide patient care. In this qualitative descriptive study, ten operating room nurses working in Eastern Canada participated in open-ended, individual audiotaped interviews that were transcribed for analysis using Boyatzis' method for code development. Three categories of factors contributing to abuse were developed. The first, culture of the OR, included environment and hierarchy. The second, catalysts of abuse, included nurses' positions and experience as well as non-nurse factors such as resources and interpersonal relationships among physicians. The third category, perceived effects, included psychological, physical and social health consequences for nurses. Effects on patient care consisted of safety and potential challenges to access. Nursing practice implications included mentoring, support and accountability for action. Educational implications related to interdisciplinary education and increased education on communication, assertiveness, and awareness of abuse. Implications for research included studying perceptions of other health-care providers including physicians, studying recruitment and retention in relation to abuse, and studying other abuse in health care such as horizontal violence. We suggest a proactive approach for empowering OR nurses to address abuse and an increased focus on interdisciplinary roles.


Structure of the rat gene encoding cholesterol 7 alpha-hydroxylase.
Cholesterol 7 alpha-hydroxylase (7 alpha-hydroxylase) is a microsomal cytochrome P-450 that catalyzes the first and rate-limiting step in bile acid biosynthesis, the major catabolic pathway in cholesterol homeostasis. The gene encoding the rat 7 alpha-hydroxylase has been isolated and characterized. Southern blotting experiments demonstrated that the gene is present in a single copy in the rat genome. DNA sequence analysis showed that the 7 alpha-hydroxylase gene is unique among the characterized cytochrome P-450s in that it contains only six exons. Nuclease S1 and primer-extension mapping experiments positioned the 5'-ends of the 7 alpha-hydroxylase mRNA approximately 20-25 nucleotides downstream of a consensus TATAAA sequence. RNA blotting experiments demonstrated the presence of multiple 7 alpha-hydroxylase mRNAs that differ in the lengths of their 3'-untranslated regions.


Six-loop Konishi anomalous dimension from the Y system.
We compute the Konishi anomalous dimension perturbatively up to six loop using the finite set of functional equations derived recently in Gromov et al. [J. High Energy Phys. 07 (2012) 023]. The recursive procedure can be, in principle, extended to higher loops, the only obstacle being the complexity of the computation.


Temporal and spatial dynamics of phytoplankton near farm fish in eutrophic reservoir in Pernambuco, Brazil.
Spatial and temporal variations in phytoplankton communities in continental waters have received attention from limnologists, since they are differently influenced by many physico-chemical and biological factors. This study was undertaken with the aim to identify the environmental variables that influence the temporal and spatial dynamics of the phytoplankton near a fish farm in the Jucazinho reservoir, in a semi-arid region of Northeastern Brazil. Samples were taken from three sampling sites, at two depths during the rainy (Aug 2008, Feb and Mar 2009) and dry (Oct, Nov and Dec 2008) seasons. Phytoplankton was identified, density determined, and biomass values obtained. Concomitantly, abiotic analyses were performed for the characterization of the system. The reservoir was homogeneous with regard to the spatial-temporal variation in hydrological variables: water well oxygenated at the surface and anoxic at the bottom; pH ranging from neutral to alkaline; temperatures always above 25 degreeC; high turbidity; and high electrical conductivity at all sampling sites and both depths. For both seasons, there was limited nitrogen and high concentrations of phosphorus. Cyanophyta species were predominant, generally representing 80% of the phytoplankton biomass throughout practically the entire study, at all sampling sites and both depths. Co-dominance of cyanobacteria belonging to H1, MP, S1 and Sn associations was recorded in most of the months studied, except August 2008, when there was a substitution of the S1 association (Planktothrix agardhii) by the P association (Aulacoseira granulata). Water temperature, precipitation and pH were the parameters with the greatest influence over the temporal variation in phytoplankton, whereas the vertical distribution of the phytoplankton biomass was directly related to the availability of light in the water column. There were no spatial or temporal differences in water quality, likely due to the fact that the sampling sites were near to one another and received the same nutrients stemming from the rations given to the fish as well as their excrement. The predominance of cyanobacteria through the study was certainly due to the influence of the feeding products offered to and excreted from the fish, which eutrophicated the system. In temporal terms, the change in the algal structure is explained by the change in the physical conditions of the water between the surface and bottom, as well as the climatologic conditions, especially the change in wind direction, with the consequent fetch of the water.


Twelve tips for using applied improvisation in medical education.
Future physicians will practice medicine in a more complex environment than ever, where skills of interpersonal communication, collaboration and adaptability to change are critical. Applied improvisation (or AI) is an instructional strategy which adapts the concepts of improvisational theater to teach these types of complex skills in other contexts. Unique to AI is its very active teaching approach, adapting theater games to help learners meet curricular objectives. In medical education, AI is particularly helpful when attempting to build students' comfort with and skills in complex, interpersonal behaviors such as effective listening, person-centeredness, teamwork and communication. This article draws on current evidence and the authors' experiences to present best practices for incorporating AI into teaching medicine. These practical tips help faculty new to AI get started by establishing goals, choosing appropriate games, understanding effective debriefing, considering evaluation strategies and managing resistance within the context of medical education.


Detailed kinetic analysis of a family 52 glycoside hydrolase: a beta-xylosidase from Geobacillus stearothermophilus.
Geobacillus stearothermophilus T-6 encodes for a beta-xylosidase (XynB2) from family 52 of glycoside hydrolases that was previously shown to hydrolyze its substrate with net retention of the anomeric configuration. XynB2 significantly prefers substrates with xylose as the glycone moiety and exhibits a typical bell-shaped pH dependence curve. Binding properties of xylobiose and xylotriose to the active site were measured using isothermal titration calorimetry (ITC). Binding reactions were enthalpy driven with xylobiose binding more tightly than xylotriose to the active site. The kinetic constants of XynB2 were measured for the hydrolysis of a variety of aryl beta-D-xylopyranoside substrates bearing different leaving groups. The Brønsted plot of log k(cat) versus the pK(a) value of the aglycon leaving group reveals a biphasic relationship, consistent with a double-displacement mechanism as expected for retaining glycoside hydrolases. Hydrolysis rates for substrates with poor leaving groups (pK(a) > 8) vary widely with the aglycon reactivity, indicating that, for these substrates, the bond cleavage is rate limiting. However, no such dependence is observed for more reactive substrates (pK(a) < 8), indicating that in this case hydrolysis of the xylosyl-enzyme intermediate is rate limiting. Secondary kinetic isotope effects suggest that the intermediate breakdown proceeds with modest oxocarbenium ion character at the transition state, and bond cleavage proceeds with even lower oxocarbenium ion character. Inhibition studies with several gluco analogue inhibitors could be measured since XynB2 has low, yet sufficient, activity toward 4-nitrophenyl beta-D-glucopyranose. As expected, inhibitors mimicking the proposed transition state structure, such as 1-deoxynojirimycin, bind with much higher affinity to XynB2 than ground state inhibitors.


Ischemic Heart Disease Selectively Modifies the Right Atrial Appendage Transcriptome.
Background: Although many pathological changes have been associated with ischemic heart disease (IHD), molecular-level alterations specific to the ischemic myocardium and their potential to reflect disease severity or therapeutic outcome remain unclear. Currently, diagnosis occurs relatively late and evaluating disease severity is largely based on clinical symptoms, various imaging modalities, or the determination of risk factors. This study aims to identify IHD-associated signature RNAs from the atrial myocardium and evaluate their ability to reflect disease severity or cardiac surgery outcomes. Methods and Results: We collected right atrial appendage (RAA) biopsies from 40 patients with invasive coronary angiography (ICA)-positive IHD undergoing coronary artery bypass surgery and from 8 patients ICA-negative for IHD (non-IHD) undergoing valvular surgery. Following RNA sequencing, RAA transcriptomes were analyzed against 429 donors from the GTEx project without cardiac disease. The IHD transcriptome was characterized by repressed RNA expression in pathways for cell-cell contacts and mitochondrial dysfunction. Increased expressions of the CSRNP3, FUT10, SHD, NAV2-AS4, and hsa-mir-181 genes resulted in significance with the complexity of coronary artery obstructions or correlated with a functional cardiac benefit from bypass surgery. Conclusions: Our results provide an atrial myocardium-focused insight into IHD signature RNAs. The specific gene expression changes characterized here, pave the way for future disease mechanism-based identification of biomarkers for early detection and treatment of IHD.


Insoluble fibronectin activates the Na/H antiporter by clustering and immobilizing integrin alpha 5 beta 1, independent of cell shape.
Growth of anchorage-dependent cells requires both soluble mitogens and insoluble extracellular matrix molecules such as fibronectin. Soluble growth factors activate chemical signaling pathways and stimulate proliferation by binding to transmembrane receptors. Insoluble fibronectin also binds to cell-surface receptors; however, it is thought to act primarily via effects on the cytoskeleton and cell shape. We recently demonstrated that cell spreading on surface-adsorbed fibronectin activates the Na/H antiporter and that inhibition of this chemical-signaling pathway suppresses growth. We now show that insoluble fibronectin activates the Na/H antiporter by clustering and immobilizing integrin alpha 5 beta 1, independent of effects on cell shape. These results show that an extracellular matrix receptor can behave similarly to a growth factor receptor to activate a signaling pathway implicated in growth control.


The early evolution of spinal cord lesions on MR imaging following traumatic spinal cord injury.
How early spinal cord injury (SCI) lesions evolve in patients after injury is unknown. The purpose of this study was to characterize the early evolution of spinal cord edema and hemorrhage on MR imaging after acute traumatic SCI. We performed a retrospective analysis of 48 patients with clinically complete cervical spine injury. Inclusion criteria were the clear documentation of the time of injury and MR imaging before surgical intervention within 72 hours of injury. The length of intramedullary spinal cord edema and hemorrhage was assessed. The correlation between time to imaging and lesion size was determined by multiple regression analysis. Short-interval follow-up MR imaging was also available for a few patients (n = 5), which allowed the direct visualization of changes in spinal cord edema. MR imaging demonstrated cord edema in 100% of patients and cord hemorrhage in 67% of patients. The mean longitudinal length of cord edema was 10.3 +/- 4.0 U, and the mean length of cord hemorrhage was 2.6 +/- 2.0 U. Increased time to MR imaging correlated to increased spinal cord edema length (P = .002), even after accounting for the influence of other variables. A difference in time to MR imaging of 1.2 days corresponded to an average increase in cord edema by 1 full vertebral level. Hemorrhage length was not affected by time to imaging (P = .825). A temporal increase in the length of spinal cord edema was confirmed in patients with short-interval follow-up MR imaging (P = .003). Spinal cord edema increases significantly during the early time period after injury, whereas intramedullary hemorrhage is comparatively static.


Preliminary experience with delayed non-operative therapy of multiple hand and wrist contractures in a woman with Freeman-Sheldon syndrome, at ages 24 and 28 years.
We describe two proof-of-concept trials of delayed non-operative therapy of multiple hand and wrist contractures in a woman with a severe expression of Freeman-Sheldon syndrome (FSS), at ages 24 and 28 years. Having presented as an infant to a university referral centre, passive correction was not accompanied by strengthening exercises, and correction was lost. FSS is described as a myopathic distal arthrogryposis; diagnosis requires the following: microstomia, whistling face appearance, H-shaped chin dimpling, nasolabial folds, and multiple hand and foot contractures. Spinal deformities, metabolic and gastroenterological problems, other craniofacial characteristics, and visual and auditory impairments, are frequent findings. To avoid possible FSS-associated complications of malignant hyperthermia and difficult intubation, and to reduce or eliminate need for surgery, we proceeded with passive manipulation without anaesthesia or sedation. We believe this is the first report of attempted non-operative correction of multiple hand and wrist contractures in an adult with FSS.


Humoral immune response profile of a cattle herd vaccinated with 5- and 10-times Bakirköy strain sheep pox vaccine under field conditions.
Vaccination is the most effective control measure for Lumpy Skin Disease (LSD). The Bakırköy strain-derived sheep pox vaccine (SPPV) has been used against LSD in Türkiye since 2013. In this study, a cattle herd was vaccinated with SPPV and 35 cattle, of which 9 and 26 received 10 and 5 sheep doses, respectively, were followed for 200 days for humoral immune responses. Additionally, maternal antibodies in colostrum-fed calves were investigated. The humoral immune responses of naive and previously vaccinated cattle were compared to determine the effects of annual re-vaccination. Furthermore, the compatibility of the VNT and ELISA tests was analyzed. According to the results, on day 30 post-vaccination, 19 and 13 out of 35 cattle were positive for VNT and ELISA, respectively. The number of seropositive cattle was higher in the group that had been vaccinated in previous years than in naive cattle. No significant differences were observed in the number of positive cattle between the groups vaccinated with the 5- and 10- doses. In colostrum-fed calves grouped according to age, the seropositivity rate was 87 % (41/47) in the one-week-old group, while this rate was only 18 % (3/16) in the 3-month-old group. It was determined that vaccination at different stages in the last four months of pregnancy did not cause any difference in the number of seropositive calves in one-week-old calves fed with colostrum. The concordance between VNT and ELISA tests was lower in 5-dose vaccinated group than 10-dose vaccinated and colostrum-fed calves groups. This study provides insights into the effect of the vaccination strategy followed by Türkiye during its combat of LSD and revealed that annual repeated vaccination using heterologous vaccine has significant positive effects on humoral immun response at the herd level.


On the quasi-isometric and bi-Lipschitz classification of 3D Riemannian Lie groups.
This note is concerned with the geometric classification of connected Lie groups of dimension three or less, endowed with left-invariant Riemannian metrics. On the one hand, assembling results from the literature, we give a review of the complete classification of such groups up to quasi-isometries and we compare the quasi-isometric classification with the bi-Lipschitz classification. On the other hand, we study the problem whether two quasi-isometrically equivalent Lie groups may be made isometric if equipped with suitable left-invariant Riemannian metrics. We show that this is the case for three-dimensional simply connected groups, but it is not true in general for multiply connected groups. The counterexample also demonstrates that 'may be made isometric' is not a transitive relation.


SAXS and stability studies of iron-induced oligomers of bacterial frataxin CyaY.
Frataxin is a highly conserved protein found in both prokaryotes and eukaryotes. It is involved in several central functions in cells, which include iron delivery to biochemical processes, such as heme synthesis, assembly of iron-sulfur clusters (ISC), storage of surplus iron in conditions of iron overload, and repair of ISC in aconitase. Frataxin from different organisms has been shown to undergo iron-dependent oligomerization. At least two different classes of oligomers, with different modes of oligomer packing and stabilization, have been identified. Here, we continue our efforts to explore the factors that control the oligomerization of frataxin from different organisms, and focus on E. coli frataxin CyaY. Using small-angle X-ray scattering (SAXS), we show that higher iron-to-protein ratios lead to larger oligomeric species, and that oligomerization proceeds in a linear fashion as a results of iron oxidation. Native mass spectrometry and online size-exclusion chromatography combined with SAXS show that a dimer is the most common form of CyaY in the presence of iron at atmospheric conditions. Modeling of the dimer using the SAXS data confirms the earlier proposed head-to-tail packing arrangement of monomers. This packing mode brings several conserved acidic residues into close proximity to each other, creating an environment for metal ion binding and possibly even mineralization. Together with negative-stain electron microscopy, the experiments also show that trimers, tetramers, pentamers, and presumably higher-order oligomers may exist in solution. Nano-differential scanning fluorimetry shows that the oligomers have limited stability and may easily dissociate at elevated temperatures. The factors affecting the possible oligomerization mode are discussed.


Are inhaled longacting β2 agonists detrimental to asthma?
Possible adverse effects of adrenergic bronchodilators in asthma have been the subject of discussion for more than half a century, with recent intense debate about the safety of longacting β agonists (LABAs). In this Debate, we consider the issues of bronchodilator and bronchoprotective tolerance resulting from the frequent use of bronchodilators, which is noted particularly with shortacting drugs, but has also been shown to occur quicker and to a greater extent with LABAs. Increased allergen responsiveness and masking allowing inflammation to increase, while symptoms and lung function remain apparently controlled, have also been observed. Studies in which LABAs were used as monotherapy were associated with increased mortality. However, several studies have shown the benefits of adding LABAs to inhaled corticosteroids (ICS). Meta-analyses of asthma clinical trials involving LABAs showed that, when given with mandatory ICS, LABAs were not associated with an increased risk of death, intubations, or hospital admission for exacerbations when compared with use of the same dose of ICS only. Withdrawal of LABA therapy once symptom control is achieved is often associated with subsequent loss of symptom control. When used for appropriate indications, LABAs should be combined with ICS in one inhaler so that monotherapy is not possible.


Acute psychological impact of coronavirus disease 2019 outbreak among psychiatric professionals in China: a multicentre, cross-sectional, web-based study.
To assess the magnitude of mental health outcomes and associated factors among psychiatric professionals in mental health services during COVID-19 in China. This cross-sectional, survey-based, region-stratified study collected demographic data and mental health measurements from psychiatric professionals in 34 hospitals between 29 January and 7 February 2020, in China. Hospitals equipped with fever clinics or deployed on wards for patients with COVID-19 were eligible. The severity of symptoms of depression, anxiety, insomnia and distress were assessed by the Chinese versions of 9-item Patient Health Questionnaire, 7-item Generalised Anxiety Disorder, 7-item Insomnia Severity Index and 22-item Impact of Event Scale-Revised, respectively. Multivariable logistic regression and structural equation modelling was performed to identify factors associated with mental health outcomes. A total of 610 psychiatric professionals were included. 29.8% were employed in Wuhan, and 22.5% were frontline workers. A considerable proportion of participants reported symptoms of depression (461 (75.6%)), anxiety (282 (46.2%)), insomnia (336 (55.1%)) and mental stress (481 (78.9%)). Psychiatric symptoms were associated with worrying about infection (eg, OR 2.36 (95% CI 1.27 to 4.39) for anxiety), risks of exposure to COVID-19 (eg, having inadequate personal protection equipment, OR 2.43 (1.32 to 4.47) for depression) and self-perceived physical health (eg, OR 3.22 (2.24 to 4.64) for mental stress). Information sources of COVID-19 were also found to be both positively (eg, information from relatives, OR 2.16 (1.46 to 3.21) for mental stress) and negatively (eg, information from TV, OR 0.52 (0.35 to 0.77) for mental stress) associated with mental stress. There is preliminary evidence that mental health might benefit from greater availability of mental healthcare services. The structural equation model analysis indicated that worrying about infection may be the primary mediator via which risk of exposure to COVID-19 pandemic affects the mental health of psychiatric professionals. The current findings demonstrate several pathways via which the COVID-19 pandemic may have negatively affected the mental health of psychiatric professionals in China.


Esophageal pressure measurements under different conditions of intrathoracic pressure. An in vitro study of second generation balloon catheters.
The aim of this study was to evaluate in vitro the accuracy of second generation esophageal catheters at different surrounding pressures and filling volumes and to suggest appropriate catheter management in clinical practice. Six different esophageal catheters were placed in an experimental chamber at four chamber pressures (0, 10, 20 and 30 cmH2O) and at filling volumes ranging from 0 to 10 mL. The working volume was defined as the volume range between the maximum (Vmax) and minimum (Vmin) volumes achieving acceptable accuracy (defined by a balloon transmural pressure ± 1 cmH2O). Accuracy was evaluated for a standard volume of 0.5 mL and for volumes recommended by manufacturers. Data are shown as median and interquartile range. In the four conditions of chamber pressure Vmin, Vmax and working volume were 1.0 (0.5, 1.5), 5.3 (3.8, 7.1), and 3.5 (2.9, 6.1) mL. Increasing chamber pressure increased Vmin (rho=0.9; P<0.0001), that reached 2.0 mL (1.6-2.0) at 30 cmH2O. Vmax and working volumes differed among catheters, whereas Vmin did not. By injecting 0.5 mL and the minimum recommended volume by manufacturer, balloon transmural pressure was <-1 cmH2O in 71% and 53% of cases, it was negatively related to chamber pressure (rho=-0.97 and -0.71; P<0.0001) and reached values of -10.4 (-12.4, -9.7) and -9.8 (-10.6, -3.4) at 30 cmH2O. Measuring positive esophageal pressures needs higher injected volumes than usually recommended. The range of appropriate filling volumes is catheter-specific. Both absolute values and respiratory changes of esophageal pressure can be underestimated by an underfilled balloon.


Distribution of nociceptin/orphanin FQ precursor protein and receptor in brain and spinal cord: a study using in situ hybridization and X-gal histochemistry in receptor-deficient mice.
Nociceptin/orphanin FQ (N/OFQ) is an opioid-like heptadecapeptide agonist for the opioid receptor homolog, N/OFQ receptor. To explore the precise distribution of the peptide-receptor system, the authors examined the brain and spinal cord from receptor-deficient mice bearing the targeted mutation (morc(m1)), a lacZ insertional mutation in the N/OFQ receptor gene. Precursor protein N/OFQ (preproN/OFQ) mRNA was detected by using in situ hybridization, and the N/OFQ receptor was detected by using X-gal histochemistry. The N/OFQ receptor reflected by lacZ expression was observed at high levels in the dentate gyrus, lateral septum, subparafascicular thalamic nucleus, medial preoptic area, median preoptic nucleus, ventromedial preoptic nucleus, anterior hypothalamic area, paraventricular hypothalamic nucleus, ventromedial hypothalamic nucleus, auditory brainstem nuclei, pontine dorsal tegmentum, and nucleus of the solitary tract. In situ detection of the N/OFQ receptor mRNA by digoxigenin-labeled riboprobes coupled with tyramide signal amplification in normal and wild-type mice resulted in the regional distribution paralleling the lacZ expression in these regions. PreproN/OFQ mRNA was expressed at high levels in the subparafascicular thalamic nucleus, central gray, central tegmental field, auditory brainstem nuclei, caudal spinal trigeminal nucleus, and spinal dorsal horn. Furthermore, variable levels of expression of the peptide and receptor were seen in distinct sites of the brain and spinal cord. These data indicate a correspondence of the peptide and the receptor in local distribution at limbic, hypothalamic, and brainstem sites. Together with concurrent physiologic and behavioral studies in mutant mice, the results suggest functional roles for the N/OFQ system, including the central regulation of learning and memory, hearing ability, water balance, food intake, and blood pressure.


Resolving the paradox for protein aggregation diseases: a common mechanism for aggregated proteins to initially attack membranes without needing aggregates.
Paradoxically, aggregation of specific proteins is characteristic of many human diseases and aging, yet aggregates have been found to be unnecessary for initiating pathogenesis. Here we determined the NMR topology and dynamics of a helical mutant in a membrane environment transformed from the 125-residue cytosolic all-β MSP by the ALS-causing P56S mutation. Unexpectedly, despite its low hydrophobicity, the P56S major sperm protein (MSP) domain becomes largely embedded in the membrane environment with high backbone rigidity. Furthermore it is composed of five helices with amphiphilicity comparable to those of the partly-soluble membrane toxin mellitin and α-synuclein causing Parkinson's disease. Consequently, the mechanism underlying this chameleon transformation becomes clear: by disrupting the specific tertiary interaction network stabilizing the native all-β MSP fold to release previously-locked amphiphilic segments, the P56S mutation acts to convert the classic MSP fold into a membrane-active protein that is fundamentally indistinguishable from mellitin and α-synuclein which are disordered in aqueous solution but spontaneously partition into membrane interfaces driven by hydrogen-bond energetics gained from forming α-helix in the membrane environments. As segments with high amphiphilicity exist in all proteins, our study successfully resolves the paradox by deciphering that the proteins with a higher tendency to aggregate have a stronger potential to partition into membranes through the same mechanism as α-synuclein to initially attack membranes to trigger pathogenesis without needing aggregates. This might represent the common first step for various kinds of aggregated proteins to trigger familiar, sporadic and aging diseases. Therefore the homeostasis of aggregated proteins in vivo is the central factor responsible for a variety of human diseases including aging. The number and degree of the membrane attacks by aggregated proteins may act as an endogenous clock to count down the aging process. Consequently, a key approach to fight against them is to develop strategies and agents to maintain or even enhance the functions of the degradation machineries.


Plasmonic bandpass filters with cascaded rectangular ring resonators.
We theoretically present the analysis and design of a nanoplasmonic bandpass filter with flat-top spectral characteristics by cascading a series of directly connected rectangular ring resonators based on metal-insulator-metal waveguides. Analyzed by the equivalent lumped circuit model of the transmission line to plasmonic waveguides, the transmission properties of a symmetric rectangular ring resonator with the directly connected input and output waveguides are approximately the same as that of a Fabry-Perot resonator. Then the thin-film design methodology is applied to realize a plasmonic bandpass filter with the squared passband. An example of cascaded two-rectangular ring resonator structure is numerically demonstrated by using the transmission line model and 2D finite difference time domain method.


Targeted transduction patterns in the mouse brain by lentivirus vectors pseudotyped with VSV, Ebola, Mokola, LCMV, or MuLV envelope proteins.
Lentiviral vectors have proven to be promising tools for transduction of central nervous system (CNS) cells in vivo and in vitro. In this study, CNS transduction patterns of lentiviral vectors pseudotyped with envelope glycoproteins from Ebola virus, murine leukemia virus (MuLV), lymphocytic choriomeningitis virus (LCMV), or the rabies-related Mokola virus were compared to a vector pseudotyped with the vesicular stomatitis virus glycoprotein (VSV-G). Mokola-, LCMV-, and VSV-G-pseudotyped vectors transduced similar populations, including striatum, thalamus, and white matter. Mokola-pseudotyped vectors were the most efficient of the three. MuLV-pseudotyped lentivirus efficiently transduced striatum and hippocampal dentate gyrus. In contrast, no transduction resulted from injection of Ebola-pseudotyped virus in the CNS. The same pattern was observed in vitro with primary cultured oligodendrocytes. LCMV, MuLV, and Ebola pseudotypes were the most stable. These results demonstrate that targeted transduction in the CNS can be achieved using specific envelope glycoproteins to pseudotype lentiviral vectors, and support the use of Mokola-pseudotyped and MuLV-pseudotyped lentiviral vectors as efficient and stable alternatives to VSV-G-pseudotyped vectors for experiments in the mouse CNS.


Ionic-liquid-catalyzed efficient transformation of γ-valerolactone to methyl 3-pentenoate under mild conditions.
Green nylons! Acidic ionic-liquid catalysis for the transformation of γ-valerolactone into methyl 3-pentenoate (M3P) is shown to be performed efficiently under mild conditions. M3P is obtained selectively from a reaction at 170 °C for 3.5 h in the presence of an acidic ionic liquid that has a low vapor pressure, high thermal stability, and excellent catalytic performance. A possible reaction pathway for this conversion is also presented.


Waveguide mode amplitude estimation using warping and phase compensation.
In shallow water, low-frequency propagation can be described by modal theory. Acoustical oceanographic measurements under this situation have traditionally relied on spatially filtering signals with arrays of synchronized hydrophones. Recent work has demonstrated how a method called warping allows isolation of individual mode arrivals on a single hydrophone, a discovery that subsequently opened the door for practical single-receiver source localization and geoacoustic inversion applications. Warping is a non-linear resampling of the signal based on a simplistic waveguide model. Because warping is robust to environmental mismatch, it provides accurate estimates of the mode phase even when the environment is poorly known. However, the approach has issues with mode amplitude estimation, particularly for the first arriving mode. As warping is not invariant to time shifting, it relies on accurate estimates of the signal's time origin, which in turn heavily impacts the first mode's amplitude estimate. Here, a revised warping operator is proposed that incorporates as much prior environmental information as possible, and is actually equivalent to compensating the relative phase of each mode. Warping and phase compensation are applied to both simulated and experimental data. The proposed methods notably improve the amplitude estimates of the first arriving mode.


Articular cartilage friction increases in hip joints after the removal of acetabular labrum.
The acetabular labrum is believed to have a sealing function. However, a torn labrum may not effectively prevent joint fluid from escaping a compressed joint, resulting in impaired lubrication. We aimed to understand the role of the acetabular labrum in maintaining a low friction environment in the hip joint. We did this by measuring the resistance to rotation (RTR) of the hip, which reflects the friction of the articular cartilage surface, following focal and complete labrectomy. Five cadaveric hips without evidence of osteoarthritis and impingement were tested. We measured resistance to rotation of the hip joint during 0.5, 1, 2, and 3 times body weight (BW) cyclic loading in the intact hip, and after focal and complete labrectomy. Resistance to rotation, which reflects articular cartilage friction in an intact hip was significantly increased following focal labrectomy at 1-3 BW loading, and following complete labrectomy at all load levels. The acetabular labrum appears to maintain a low friction environment, possibly by sealing the joint from fluid exudation. Even focal labrectomy may result in increased joint friction, a condition that may be detrimental to articular cartilage and lead to osteoarthritis.


Rho GTPases regulate axon growth through convergent and divergent signaling pathways.
Rho GTPases are essential regulators of cytoskeletal reorganization, but how they do so during neuronal morphogenesis in vivo is poorly understood. Here we show that the actin depolymerization factor cofilin is essential for axon growth in Drosophila neurons. Cofilin function in axon growth is inhibited by LIM kinase and activated by Slingshot phosphatase. Dephosphorylating cofilin appears to be the major function of Slingshot in regulating axon growth in vivo. Genetic data provide evidence that Rho or Rac/Cdc42, via effector kinases Rok or Pak, respectively, activate LIM kinase to inhibit axon growth. Importantly, Rac also activates a Pak-independent pathway that promotes axon growth, and different RacGEFs regulate these distinct pathways. These genetic analyses reveal convergent and divergent pathways from Rho GTPases to the cytoskeleton during axon growth in vivo and suggest that different developmental outcomes could be achieved by biases in pathway selection.


Risk and protective factors for leprosy development determined by epidemiological surveillance of household contacts.
Household contacts of leprosy patients are the group with the highest risk of developing the disease, and although many risk or prevention factors have been identified, they have not been employed in leprosy-monitoring programs. This investigation aimed to establish the relative risks or the preventive effects of the presence of BCG vaccination, the Mitsuda test, and the ML-Flow assay. Household contacts (1,396) were monitored for a 5-year period. Twenty-eight contacts (2%) developed leprosy and had their clinical and operational classifications established. All immunological tests were performed, and intradermal BCG vaccination was given after the BCG scar count. Of the affected contacts, 75% developed the disease in the first year, and 71.4% were classified as having paucibacillary forms. Contacts of lepromatous leprosy patients presented a 3.8-fold-higher risk of developing leprosy. BCG vaccination and the Mitsuda test showed a protective effect against leprosy of 0.27 (at least one scar) and 0.16 (>7 mm), respectively, and the positive ML-Flow test indicated a relative risk approximately sixfold higher for occurrence of the disease. All unfavorable combinations of two and three assays generated significant risk values that ranged from 5.76 to 24.47, with the highest risk given by the combination of no BCG scar, negative Mitsuda test, and positive ML-Flow test. We suggest that the BCG vaccination may be given to stimulate Mitsuda test positivity, reducing the patient's risk of developing multibacillary forms. The high significance of these tests may have a great impact on programs to monitor contacts and should be used to improve early detection and treatment.


Incidence of silent lacunar lesion in normal adults and its relation to cerebral blood flow and risk factors.
There are few reports comparing incidence of silent lacunar lesions detected by magnetic resonance imaging and cerebral blood flow in neurologically normal adults. We studied the incidence of such lesions and its relation to cerebral blood flow and risk factors in 246 neurologically normal adults (145 men, mean age 62 years; 101 women, mean age 60 years) who received health screening examinations of the brain. Thirty-two subjects (13%) had possible silent lacunar lesions (66% of these were recognized by both T1- and T2-weighted image). The regional cerebral blood flow measured by the xenon-133 inhalation method was significantly lower in subjects with silent lacunes than in those without (p less than 0.02). Cerebral blood flow was mildly but significantly decreased in those with silent lacunes (p less than 0.05). Periventricular hyperintensity was closely related to silent lacune (p less than 0.01). However, there was no significant difference in cerebral blood flow between subjects with and without apparent periventricular hyperintensity. Silent lacunar lesion was closely related to decrease of cerebral circulation and may be an important risk factor for symptomatic cerebrovascular disease.


An Unnecessary Russell's Viper Bite on the Tongue Due to Live Snake Worship and Dangerous First Aid Emphasise the Urgent Need for Stringent Policies.
India suffers the highest incidence of snakebite envenomation (SBE) in the world. Rural communities within India and other countries have long-held cultural beliefs surrounding snakes and SBE treatments, with snake statues present in numerous Hindu temples. While most cultural beliefs are well respected and do not affect anyone, some people worship live venomous snakes without any safety precautions. Moreover, they practice various inappropriate first aid and traditional treatments that exacerbate SBE-induced complications. We report an unusual case of SBE on the tongue of a patient who was bitten while worshipping Russell's viper following the advice of an astrologer based on the appearance of a snake in the patient's dream. Following the bite, the tongue was deeply incised by the priest as a first aid to mitigate SBE-induced complications. The patient suffered profuse bleeding and swelling of the tongue resulting in difficulties in intubating them. The patient regained consciousness after antivenom administration, intranasal ventilation, and blood removal from the mouth. The tongue underwent extensive surgery to restore movement and function. This report advises caution to those undertaking the extremely risky practice of worshipping live snakes and emphasises the urgent need to develop and enforce policies to mitigate such actions and educate rural communities.


Comparing attentional bias to smoking cues in current smokers, former smokers, and non-smokers using a dot-probe task.
Much evidence documents that individuals with emotional and drug-use disorders demonstrate biased attention toward stimuli associated with their disorder. This bias appears to diminish following successful treatment. Two studies examined whether current cigarette smokers show biased attention toward smoking-related images compared with non-smokers (Studies 1 and 2) and whether this bias is less pronounced in former smokers (Study 2). Attentional bias toward cigarette-related photographs was examined using the dot-probe task. Pairs of images (one smoking-related) appeared side by side for 500 ms on a computer screen prior to the presentation of a probe (an asterisk) replacing one of the photographs. Subjects struck a key as quickly as possible to indicate the probe location. Attentional bias was defined as faster reaction times when the probe replaced the smoking-related image. In both studies, current smokers displayed significantly greater attentional bias toward cigarette stimuli than did non-smokers. Former smokers in Study 2 displayed an intermediate level of bias, but did not differ significantly in bias score from either of the other groups. These results support further use of the dot-probe task as a measure of attentional bias in non-abstinent smokers and in individuals undergoing smoking cessation treatment.


Appropriate Nitrogen form Ratio and UV-A Supplementation Increased Quality and Production in Purple Lettuce (Lactuca sativa L.).
Purple lettuce (Lactuca sativa L. cv. Zhongshu Purple Lettuce) was chosen as the trial material, and LED intelligent light control consoles were used as the light sources. The purpose was to increase the yield and quality of purple lettuce while lowering its nitrate level. By adding various ratios of NO3--N and NH4+-N to the nutrient solution and 20 µmol m-2 s-1 UV-A based on white, red, and blue light (130, 120, 30 µmol m-2 s-1), the effects of different NO3--N/NH4+-N ratios (NO3--N, NO3--N/NH4+-N = 3/1, NH4+-N) and UV-A interaction on yield, quality, photosynthetic characteristics, anthocyanins, and nitrogen assimilation of purple lettuce were studied. In order to produce purple lettuce hydroponically under controlled environmental conditions, a theoretical foundation and technological specifications were developed, taking into account an appropriate UV-A dose and NO3--N/NH4+-N ratio. Results demonstrate that adding a 20 µmol m-2 s-1 UV-A, and a NO3--N/NH4+-N treatment of 3/1, significantly reduced the nitrate level while increasing the growth, photosynthetic rate, chlorophyll, carotenoid, and anthocyanin content of purple lettuce. The purple leaf lettuce leaves have an enhanced capacity to absorb nitrogen. Furthermore, plants have an acceleration of nitrogen metabolism, which raises the concentration of free amino acids and soluble proteins and promotes biomass synthesis. Thus, based on the NO3--N/NH4+-N (3/1) treatment, adding 20 µmol m-2 s-1 UV-A will be helpful in boosting purple lettuce production and decreasing its nitrate content.


Involvement of protein radical, protein aggregation, and effects on NO metabolism in the hypochlorite-mediated oxidation of mitochondrial cytochrome c.
Cytochrome c (cyt c)-derived protein radicals, radical adduct aggregates, and protein tyrosine nitration have been implicated in the pro-apoptotic event connecting inflammation to the development of diseases. During inflammation, one of the reactive oxygen species metabolized via neutrophil activation is hypochlorite (HOCl); destruction of the mitochondrial electron transport chain by hypochlorite is considered to be a damaging factor. Previous study has shown that HOCl induces the site-specific oxidation of cyt c at met-80. In this work, we have assessed the hypothesis that exposure of cyt c to physiologically relevant concentrations of HOCl leads to protein-derived radical and consequent protein aggregation, which subsequently affects cyt c's regulation of nitric oxide metabolism. Reaction intermediates, chemical pathways available for protein aggregation, and protein nitration were examined. A weak ESR signal for immobilized nitroxide derived from the protein was detected when a high concentration of cyt c was reacted with hypochlorite in the presence of the nitroso spin trap 2-methyl-2-nitrosopropane. When a low concentration of cyt c was exposed to the physiologically relevant levels of HOCl in the presence of 5,5-dimethyl-pyrroline N-oxide (DMPO), we detected DMPO nitrone adducts derived from both protein and protein aggregate radicals as assessed by Western blot using an antibody raised against the DMPO nitrone adduct. The cyt c-derived protein radicals formed by HOCl were located on lysine and tyrosine residues, with lysine predominating. Cyt c-derived protein aggregates induced by HOCl involved primarily lysine residues and hydrophobic interaction. In addition, HOCl-oxidized cyt c (HOCl-cyt c) exhibited a higher affinity for NO and enhancement of nonenzymatic NO synthesis from nitrite reduction. Furthermore, HOCl-mediated cyt c oxidation also resulted in a significant elevation of cyt c nitration derived from either NO trapping of the cyt c-derived tyrosyl radical or cyt c-catalyzed one-electron oxidation of nitrite.


Airway pressure release ventilation.
Airway pressure release ventilation (APRV) delivers continuous positive airway pressure (CPAP) and may support ventilation simultaneously. This investigation tested whether, after acute lung injury (ALI), APRV promotes alveolar ventilation and arterial oxygenation without increasing airway pressure (Paw) above the CPAP level and without depressing cardiac function. Ten anesthetized dogs randomly received either intermittent positive-pressure ventilation (IPPV) or APRV. APRV was delivered with a continuous-flow CPAP system. Expiration occurred when a switch in the expiratory limb opened and Paw decreased to near-ambient, which decreased lung volume. After baseline data collection, ALI was induced by infusing oleic acid iv. Two hours later, IPPV and APRV were administered randomly, and data were collected. With normal lungs, APRV and IPPV achieved similar gas exchange and hemodynamic function. During ALI, arterial oxygenation was improved, and peak Paw which did not exceed the CPAP level, was lower during APRV. Similar minute ventilations were delivered by both modes but resulted in lower PaCO2 with APRV. Thus, APRV decreased physiologic deadspace ventilation. Hemodynamic status was similar during both modes. Therefore, APRV is an improved method of oxygenation and ventilatory support for patients with ALI that will allow unrestricted spontaneous ventilation and may decrease the incidence of barotrauma.


Phase separation induced by ladder-like polymer-polymer complexation.
Polymer-polymer complexation in solvent is studied using an extension of the self-consistent field theory. The model polymers are capable of forming ladder-like duplex structures. The duplex formation occurs with an abrupt change of entropy, resulting in a first-order transition. Moreover, the complexation can be stabilized by solvent-polymer interactions, instead of the usual specific binding interactions. Various types of unconventional phase diagrams are predicted. For example, phase separation with decreasing χ-parameter between duplex polymer and solvent can be induced, leading to a lower critical solution temperature (LCST) behavior. Multiphase coexistence points at which two, three, or four phases coexist are also obtained. Under certain conditions a homogeneous phase becomes unstable when the polymer chain length is decreased, in contrast to the standard Flory-Huggins theory.


[Histopathological findings in incidental appendectomies during gynecologic laparotomies and their significance for the patient].
Incidental appendectomy is a more and more frequent part of gynaecological laparotomies. Doubts about the increased post-operative morbidity and mortality have been shown to be unfounded in numerous series. Opinions on contra-indications to incidental appendectomy are divided. German language authors do not do incidental appendectomies with Caesarean Sections, operations for Ectopic pregnancies, Tubo-Ovarian Abscess and Inoperable Carcinoma. In the English speaking literature an increasing number of authors show that even under these circumstances the appendectomy is without danger. Incidental appendectomy is indicated because of the high incidence of abnormal findings at microscopic examination and because the appendix may be a potential hazard for some groups of patients especially pregnant women and elderly women. An increased mortality can be attributed to the potential site of the disease in the appendix. The possible long term complications of incidental appendectomy by adhesions or bowel obstruction are lower than the potential risk of morbidity or mortality with the appendix in situ. In our series microscopic examination of one hundred and thirteen incidentally removed appendices showed abnormal findings in 82.9% of the cases. The appendices were removed in a series of 267 gynaecological laparotomies. In 17% of the cases the findings were important for the patient. An increased post-operative morbidity or mortality was not found in our series. The hospital stay was not prolonged over that of a comparitive group. The risk of a left behind appendix was calculated from a study of the literature both as to morbidity and mortality and compared to the possible short term and long term morbidity from an incidental appendectomy.


Efficacy and safety of cosibelimab, an anti-PD-L1 antibody, in metastatic cutaneous squamous cell carcinoma.
Programmed cell death receptor-1 (PD-1)-blocking antibodies are approved to treat metastatic or locally advanced cutaneous squamous cell carcinoma (CSCC) cases ineligible for curative surgery or radiation. Notwithstanding, some patients experience inadequate responses or severe immune-related adverse events (AEs), indicating the need for improved therapies. Cosibelimab is a high-affinity programmed cell death-ligand 1 (PD-L1)-blocking antibody that activates innate and adaptive immunity by blocking PD-L1 interaction with PD-1 and B7-1 receptors. It is an unmodified immunoglobulin G1 subtype with a functional Fc domain capable of inducing antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity. Here, we present results of the pivotal study of patients with metastatic CSCC from an open-label, multicenter, multiregional, multicohort, phase 1 trial of cosibelimab. In this trial, participants with metastatic CSCC received cosibelimab 800 mg intravenously every 2 weeks. Primary endpoint was objective response rate (ORR) by independent central review using Response Evaluation Criteria in Solid Tumors, V.1.1. Secondary endpoints included duration of response (DOR) and safety. Objective response was observed in 37 of 78 participants (47.4% (95% CI: 36.0% to 59.1%)), with median follow-up of 15.4 months (range: 0.4 to 40.5) as of data cut-off. Median DOR was not reached (range: 1.4+ to 34.1+ months), with response ongoing in 73.0% of participants. Common treatment-emergent AEs (≥15%) were fatigue (26.9%), rash (16.7%), and anemia (15.4%). Eighteen participants (23.1%) experienced immune-related AEs (grade 3: n=2 (2.6%); no grade 4/5). No treatment-related deaths were reported. Cosibelimab demonstrated clinically meaningful ORR and DOR and was associated with a manageable safety profile. NCT03212404.


Inhibition of cigarette smoke-induced airway secretory cell hyperplasia by indomethacin, dexamethasone, prednisolone, or hydrocortisone in the rat.
As the nonsteroidal anti-inflammatory drug indomethacin has already been shown to inhibit cigarette smoke (CS)-induced secretory cell hyperplasia in the airway epithelium of the rat, the present study was undertaken to determine the effects of prophylactic interaperitoneal injection of indomethacin (4 mg/kg body weight) compared with each of three steroidal anti-inflammatory drugs given at comparable doses. The results show that dexamethasone (given initially at 4 mg/kg), prednisolone (4 mg/kg) and hydrocortisone (4 mg/kg) were also inhibitory. Dexamethasone proved to be toxic and its dose had to be reduced to give an average dose of 2 mg/kg over the experimental period. Prednisolone and hydrocortisone were toxic only when in combination with CS. The order of descending effectiveness in inhibiting secretory cell hyperplasia was indomethacin, dexamethasone, prednisolone and hydrocortisone. Indomethacin was the most effective drug in the trachea, whilst in distal intra-pulmonary airways dexamethasone was the most effective. Depending on airway level, inhibition was between 69 and 115% of the value obtained after CS alone. The inhibition was complete in all but one instance. The results show that steroids also inhibit secretory cell hyperplasia but at a comparable dose they are generally less effective than indomethacin and may have unacceptable side effects.


The effectiveness of surgical treatment of lung cancer in Polish academic and nonacademic centers.
The theoretical advantage of academic hospitals over nonacademic are: more qualified surgeons, adequate diagnostic facilities and infrastructure, including intensive care units. The aim of the study was to compare the effectiveness of surgical lung cancer treatment in academic (ACA) and nonacademic (non-ACA) centers. This was a retrospective analysis of data from 31,777 patients surgically-treated for lung cancer during the period from 2007 to 2020 in 9 ACA and 21 non-ACA centers. The analysis considered the clinical data of patients, the effectiveness of preoperative diagnostics, the type of procedures performed, the complications, the postoperative mortality and the long-term survival. The median number of anatomical lung resection procedures was 1,218 for ACA and 550 for non-ACA centers. In the ACA group, resection using the video-assisted thoracic surgery (VATS) technique was performed significantly more often than in the non-ACA group (23.6% vs. 14.2%, P<0.001). The pN feature analysis showed significantly lower proportions of pNX (9.2%) in the ACA group than those in the non-ACA group (17.1%) (P<0.001). The rates of postoperative complications in the ACA and non-ACA groups were 30.7% and 33.8%, respectively (P<0.001). There were no significant differences in 5-year survival between the ACA and non-ACA groups (56% and 56%, respectively) (P=0.2). The present study showed that ACA centers are characterized by better preoperative diagnostics, a higher percentage of VATS lobectomies, a lower percentage of postoperative complications and a shorter hospitalization period than non-ACA centers, but there was no impact on 5-year survival.


Inactivation Mechanism of Aspergillus flavus Conidia by High Hydrostatic Pressure.
This study investigated the inactivation mechanism of Aspergillus flavus conidia by high hydrostatic pressure (HHP). Activity counts, scanning electron microscopic (SEM) analysis, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were used to study the effects of the HHP treatment on the morphology and protein composition of A. flavus spores. The results showed that that a 3-min-lasting 600 MPa treatment could completely abolish 107 colony-forming units/mL of live fungi. Furthermore, we also observed that lower spore viability corresponded to a higher Propidium Iodide absorption rate. The SEM images revealed that HHP disrupted the spore morphology and resulted in pore formation that led to the release of intracellular molecules, such as nucleic acids and proteins. The nucleic acid and protein concentration in the spore suspension increased in parallel with the increasing treatment pressure. The SDS-PAGE analysis showed that there were differences in the protein bands between the HHP-treated and untreated A. flavus spores, as the HHP treatment caused partial protein degradation and extracellular release. Therefore, the results of this study proved that high pressure could induce a morphological disruption in the internal and external cellular structures and degrade intracellular and extracellular proteins leading to an inactive state in A. flavus.


Real-time polymerase chain reaction monitoring of recombinant DNA entry into soil from decomposing roundup ready leaf biomass.
Glyphosate-tolerant, Roundup Ready (RR) soybeans account for about 57% of all genetically modified (GM) crops grown worldwide. The entry of recombinant DNA into soil from GM crops has been identified as an environmental concern due to the possibility of their horizontal transfer to soil microorganisms. RR soybeans contain recombinant gene sequences that can be differentiated from wild-type plant and microbial genes in soil by using a sequence-specific molecular beacon and real-time polymerase chain reaction (PCR). A molecular beacon-based real-time PCR system to quantify a wild-type soybean lectin ( le1) gene was designed to compare amounts of endogenous soybean genes to recombinant DNA in soil. Microcosm studies were carried out to develop methodologies for the detection of recombinant DNA from RR soybeans in soil. RR soybean leaf litterbags were imbedded in the soil under controlled environmental conditions (60% water holding capacity, 10/15 degrees C, and 8/16 h day/night) for 30 days. The soybean biomass decomposition was described using a single-phase exponential equation, and the DNA concentration in planta and in soil was quantified using real-time PCR using sequence-specific molecular beacons for the recombinant cp4 epsps and endogenous soybean lectin ( le1) genes. The biomass of RR soybean leaves was 8.6% less than nontransgenic (NT) soybean leaves after 30 days. The pooled half-disappearance time for cp4 epsps and le1 in RR and of le1 in NT soybean leaves was 1.4 days. All genes from leaves were detected in soil after 30 days. This study provides a methodology for monitoring the entry of RR and NT soybean DNA into soil from decomposing plant residues.


Assessment of interlobar variation of bronchoalveolar lavage cellular differentials in interstitial lung diseases.
Alveolitis is thought to precede permanent lung derangement in a variety of interstitial lung diseases (ILD). Bronchoalveolar lavage (BAL) cell differentials can be used to evaluate the intensity of alveolitis, whereas clinical, roentgenographic, and functional studies are insensitive monitors of lung inflammation. As ILD is generally diffuse, unilobar lavage is widely used and presumed to gauge overall lung inflammation. Consistency of lobe to lobe cell differential findings has not been systematically addressed. We analyzed 53 bilateral lobar lavages (right middle lobe and lingula) in 34 patients with sarcoidosis, idiopathic pulmonary fibrosis-collagen vascular disease (IPF-CV) and a group of mixed interstitial diseases. Cellular differentials from each lobe were independently assessed and compared. The sarcoid group, with predominantly T-lymphocytes in BAL, showed excellent interlobar correlation (r = 0.92, p less than 0.001), with only 17% showing a discrepancy of greater than 10% in percentage of T-lymphocytes. In contrast, the IPF-CV group, with predominantly neutrophils in BAL, showed good interlobar correlation (r = 0.79; p less than 0.01), but 35% had a greater than 10% discrepancy in percentage of neutrophils. Finally, the mixed group, with lymphocytes as the predominant cell in BAL, showed poor interlobar correlation (r = 0.58; p greater than 0.10), with 42% showing a greater than 10% discrepancy in percentage of lymphocytes. These results were not explicable on the basis of differences in volume of lavage fluid or total cells recovered, these parameters being remarkably consistent between lobes in the 3 patient groups and in a control group of 8 subjects without ILD. The chest roentgenogram was markedly insensitive to lavage interlobar variation.(ABSTRACT TRUNCATED AT 250 WORDS)


Sonographic diagnosis and Endo-SPONGE assisted vacuum therapy of anastomotic leakage following posterior pelvic exenteration for ovarian cancer without using a protective stoma.
Anastomotic leakage is a very significant complication after posterior pelvic exenteration and a major cause of postoperative morbidity and mortality. We present a patient who underwent an optimal debulking surgery for an advanced stage ovarian cancer (FIGO IIIC). On postoperative day 12, transvaginal ultrasound revealed an anastomotic dehiscence following an unsuspicious computer tomography scan the day before. The patient was successfully managed by transanal vacuum therapy without re-laparotomy within a period of 4 weeks after diagnosis. We conclude that high-resolution transvaginal ultrasound is a crucial method in the management of complications after surgery and even allow diagnosing leakages of colorectal anastomosis. In selected cases characterized by a small leak size and a local peritonitis confined to the pelvis a transanal vacuum therapy may avoid both surgical re-intervention and creating a secondary diverting stoma.


Chemokines and dendritic cells in inflammatory myopathies.
This review focuses on the contribution of the local production of chemokines and cytokines and of dendritic cells (DC) to the pathogenesis of inflammatory myopathies. DC are the most efficient professional antigen-presenting cells (APC), which are critical for the development of innate and adaptive immune responses. Chemokines are important mediators of the immune response as they regulate leucocyte recruitment to tissue and play a key role in inflammatory diseases by acting on T-cell and DC migration. Recent advances indicate that the muscle cell itself could participate in the inflammatory process. Furthermore, the T-helper (Th) type 1 and Th17 proinflammatory cytokines, present in myositis samples, are associated with the migration, differentiation and maturation of inflammatory cells and allow a network of interactions between all the components of the immune response. An understanding of such interactions is essential because it can lead to therapeutic applications.


Heat tolerance of a tropical-subtropical rainforest tree species Polyscias elegans: time-dependent dynamic responses of physiological thermostability and biochemistry.
Heat stress interrupts physiological thermostability and triggers biochemical responses that are essential for plant survival. However, there is limited knowledge on the speed plants adjust to heat in hours and days, and which adjustments are crucial. Tropical-subtropical rainforest tree species (Polyscias elegans) were heated at 40°C for 5 d, before returning to 25°C for 13 d of recovery. Leaf heat tolerance was quantified using the temperature at which minimal chl a fluorescence sharply rose (Tcrit ). Tcrit , metabolites, heat shock protein (HSP) abundance and membrane lipid fatty acid (FA) composition were quantified. Tcrit increased by 4°C (48-52°C) within 2 h of 40°C exposure, along with rapid accumulation of metabolites and HSPs. By contrast, it took > 2 d for FA composition to change. At least 2 d were required for Tcrit , HSP90, HSP70 and FAs to return to prestress levels. The results highlight the multi-faceted response of P. elegans to heat stress, and how this response varies over the scale of hours to days, culminating in an increased level of photosynthetic heat tolerance. These responses are important for survival of plants when confronted with heat waves amidst ongoing global climate change.


Extraordinary exposed in early motherhood - a qualitative study exploring experiences of mothers with type 1 diabetes.
Women with type 1 diabetes face several challenges during pregnancy, childbirth and in relation to breastfeeding. It is therefore of utmost importance to consider their need for specific support, early postpartum as well as in daily life after discharge from maternity care. Few studies have investigated these aspects of healthcare. The aim of this study was to explore experiences after childbirth regarding breastfeeding, glycemic control, support and well-being in women with type 1 diabetes. A hermeneutic reflective life world research approach was used in this qualitative study. Data was gathered through audio-recorded focus group discussions and individual interviews with 23 women with type 1 diabetes, 6-24 months after childbirth. After verbatim transcription, the text was analyzed in order to identify themes of meaning and a conclusive interpretation of the explored phenomenon. Experiences of extraordinary exposure challenged the women with type 1 diabetes in their transition to early motherhood. The exposure included a struggle with breastfeeding, although with a driving force to succeed. Everyday life was filled with uncertainty and unpredictability related to one's own unstable glycemic control and the women down-prioritized their own needs in favor of the child. A feeling of being disconnected from professional care further contributed to the experiences of extraordinary exposure. In early motherhood women with type 1 diabetes have a great need for support in managing daily life postpartum, which requires contemporary approaches to overlap insufficient linkage between health care professionals in maternity and child health care, and diabetes care.