Finite element analysis of zygomatic implants in intrasinus and extramaxillary approaches for prosthetic rehabilitation in severely atrophic maxillae.
To compare the extramaxillary approach with the widely used intrasinus approach via finite element method. A unilateral three-dimensional model of the craniofacial area surrounding the region of interest was developed using computed tomography image datasets. The zygomatic implants were modeled using three-dimensional computer-aided design software and virtually placed according to the described techniques together with one conventional implant and a prosthesis. The bone was assumed to be linear isotropic with a stiffness of 13.4 GPa, while the implants were of titanium alloy with a stiffness of 110 GPa. Masseter forces were applied at the zygomatic arch, and occlusal loads were applied to the surface of the prosthesis. The stresses and displacements generated on the surrounding bone and within the implant due to the simulated loading configuration were analyzed. The bone-implant interface and zygomatic implant body for the intrasinus approach produced 1.41- and 4.27-fold higher stress, respectively, compared with the extramaxillary approach under vertical loading. However, under lateral loading, the extramaxillary approach generated 2.48-fold higher stress than the intrasinus at the bone-implant interface. The zygomatic implant in the extramaxillary approach had twofold higher micromotion than those with intrasinus approach under lateral loading. No one technique was found to be superior; however, if lateral loading is used, the intrasinus approach is the most favorable for the rehabilitation of severely atrophic maxillae.


Workplace peer educators and stress.
Peer educators form an important component of company responses to HIV and AIDS. Based on interviews with peer educators working in and around a mining company in South Africa's North-West Province, the study examines the relationship between involvement in peer education and stress. The paper discusses how becoming a peer educator can be a response to the often personal stress brought about by the HIV epidemic. In addition, structural difficulties, skills deficiencies and other obstacles to effective communication with their peers can create stress. The stress that active peer education brings to individuals is discussed, particularly in regard to the embeddedness of peer educators within their communities. The need for confidentiality also magnifies stress in the case of individuals who disregard peer educators' advice. Peer educators face many stresses in managing and supporting their own lives, thus their (voluntary) work as peer educators should not be taken out of context. Using this approach, we discuss how the role of peer educator should be conceptualised and how they can be organised and supported in order that their stress be minimised and effective engagement maximised.


Fluorescent in situ hybridization with specific DNA probes offers adequate detection of Enterococcus faecalis and Enterococcus faecium in clinical samples.
Enterococcus faecalis and Enterococcus faecium are among the leading causes of hospital-acquired infections. Reliable and quick identification of E. faecalis and E. faecium is important for accurate treatment and understanding their role in the pathogenesis of infections. Fluorescent in situ hybridization (FISH) of whole bacterial cells with oligonucleotides targeted at the 16S rRNA molecule leads to a reduced time to identification. In clinical practice, FISH therefore can be used in situations in which quick identification is necessary for optimal treatment of the patient. Furthermore, the abundance, spatial distribution and bacterial cell morphology can be observed in situ. This report describes the design of two fluorescent-labelled oligonucleotides that, respectively, detect the 16S rRNA of E. faecalis and the 16S rRNA of E. faecium, Enterococcus hirae, Enterococcus mundtii, Enterococcus villorum and Enterococcus saccharolyticus. Different protocols for the application of these oligonucleotides with FISH in different clinical samples such as faeces or blood cultures are given. Enterococci in a biofilm attached to a biomaterial were also visualized. Embedding of the biomaterial preserved the morphology and therefore the architecture of the biofilm could be observed. The usefulness of other studies describing FISH for detection of enterococci is generally hampered by the fact that they have only focused on one material and one protocol to detect the enterococci. However, the results of this study show that the probes can be used both in the routine laboratory to detect and determine the enterococcal species in different clinical samples and in a research setting to enumerate and detect the enterococci in their physical environment.


Mitochondrial DNA and Parkinson's disease.
Two major lines of evidence support the hypothesis that an impairment of mitochondrial function may underlie neuronal death in Parkinson's disease. First, the neurotoxicity of the parkinsonism-inducing compound 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is due to the generation of its 1-methyl-4-phenylpyridinium (MPP+) metabolite in the central nervous system; the toxicity of MPP+ is likely to result from its ability to block mitochondrial electron flow at the level of complex I. Second, recent studies have revealed a deficiency of mitochondrial complex I activity in the brain as well as other tissues of parkinsonian patients. This enzyme activity reduction might be explained by a defect in one or more of the genes coding for the subunits of complex I. Since seven of these genes are localized in the mitochondrial genome, it is conceivable that abnormal mitochondrial DNA (mtDNA) might play a role in the pathogenesis of Parkinson's disease. The entire sequence of the human mitochondrial genome is known, and human mtDNA can be isolated and rapidly analyzed using techniques such as the polymerase chain reaction. Therefore, identification of an easily detectable mtDNA alteration might ultimately be used as a marker for the diagnosis and screening of Parkinson's disease.


ALS surrogate markers. MUNE.
Over the last decade, motor unit number estimation (MUNE) methods have been applied with increasing frequency to the study of amyotrophic lateral sclerosis. MUNE is the ideal tool for the assessment of diseases in which the primary defect is motor unit loss, as it enables quantitation and tracking of motor unit numbers while simultaneously gauging countervailing collateral reinervation. These properties make it particularly useful for assessing the effects of both neuroprotective therapies and therapies designed to enhance collateral reinervation, not only in animal models but also in the living patient. Previous studies have supplied important natural history information, confirming an average 50% loss of motor units for every six months of disease progression, and newer pathophysiological investigations are providing unique insight into motor unit behavior in the face of progressive anterior horn cell death. More recent efforts have incorporated MUNE into ongoing, multi-center clinical trials as a putative early biomarker, with encouraging results. As MUNE methods continue to be refined and disseminated, they are proving to be useful and unique tools for the study of motor neuron disease.


Feline coronavirus quantitative reverse transcriptase polymerase chain reaction on effusion samples in cats with and without feline infectious peritonitis.
Objectives The aim of the study was to determine whether feline coronavirus (FCoV) RNA in effusion samples can be used as a diagnostic marker of feline infectious peritonitis (FIP); and in FCoV RNA-positive samples to examine amino acid codons in the FCoV spike protein at positions 1058 and 1060 where leucine and alanine, respectively, have been associated with systemic or virulent (FIP) FCoV infection. Methods Total RNA was extracted from effusion samples from 20 cats with confirmed FIP and 23 cats with other diseases. Feline coronavirus RNA was detected using a reverse transcriptase quantitative polymerase chain reaction assay (qRT-PCR), and positive samples underwent pyrosequencing of position 1058 with or without Sanger sequencing of position 1060 in the FCoV spike protein. Results Seventeen (85%) of the effusion samples from 20 cats with FIP were positive for FCoV RNA, whereas none of the 23 cats with other diseases were positive. Pyrosequencing of the 17 FCoV-positive samples showed that 11 (65%) of the cats had leucine and two (12%) had methionine at position 1058. Of the latter two samples with methionine, one had alanine at position 1060. Conclusions and relevance A positive FCoV qRT-PCR result on effusions appears specific for FIP and may be a useful diagnostic marker for FIP in cats with effusions. The majority of FCoVs contained amino acid changes previously associated with systemic spread or virulence (FIP) of the virus.


First Report of Tomato spotted wilt virus in Peppers and Tomato in the Dominican Republic.
In the Dominican Republic, green bell pepper (Capsicum annuum L.) and tomato (Solanum lycopersicum L.) are widely cultivated under protected greenhouse conditions as high value commercial crops for export. For the past 2 to 3 years, pepper and tomato have been observed in protected crop facilities in Jarabacoa and Constanza in the North Region with chlorotic and necrotic spots and rings on leaves, petioles, and stems, leaf bronzing, and tip necrosis. Fruits on symptomatic pepper and tomato plants showed concentric rings, irregular chlorotic blotches and deformation, and uneven maturation and development. Incidence on pepper and tomato was 20 to 100% and 5 to 20%, respectively. In initial tests, leaves and fruits from each of 20 symptomatic tomato and pepper plants from several greenhouse facilities were reactive in Tomato spotted wilt virus (TSWV; genus Tospovirus, family Bunyaviridae) immunostrip assays (Agdia, Inc., Elkhart, IN). Since these immunostrips are known to react with other tospoviruses, such as Tomato chlorotic spot virus (TCSV) and Groundnut ring spot virus, additional molecular diagnostic assays were conducted. Leaf and fruit samples from symptomatic plants were imprinted on nitrocellulose membrane (NCM) (2), air-dried, and sent to Washington State University for confirmatory tests. Viral nucleic acids were eluted from NCM discs (1) and subjected to reverse transcription (RT)-PCR using primers gL3637 (CCTTTAACAGTDGAAACAT) and gL4435 (CATDGCRCAAGARTGRTARACAGA) designed to amplify a portion of the L RNA segment of several tospoviruses (3). A single DNA product of ~800 bp was amplified from all samples. Amplicons from two tomato (leaf and fruit) and one pepper fruit samples were cloned separately into pCR2.1 (Invitrogen Corp., Carlsbad, CA). Two independent clones per amplicon were sequenced in both orientations. Sequence analyses of these clones (GenBank Accession Nos. KF 219673 to 75) showed 100% nucleotide sequence identity among themselves and 97% identity with corresponding L RNA sequences of pepper isolates of TSWV from Taiwan (HM180088) and South Korea (HM581940), 94 to 95% with tomato isolates of TSWV from South Korea (HM581934) and Hawaii (AY070218), and 89% with a tomato isolate from Indonesia (FJ177301). These results further confirm the presence of TSWV in symptomatic tomato and pepper plants. A comparison of TSWV sequences from the Dominican Republic with TSWV isolates from the United States and other countries in the Caribbean region could not be made due to the absence of corresponding sequences of the L-RNA of the virus from these countries in GenBank. TSWV-positive samples were negative for TCSV in RT-PCR, indicating the absence of this tospovirus that has been reported in the Caribbean region (data not shown). To our knowledge, this is the first confirmed report of TSWV in tomatoes and peppers in the Dominican Republic. The presence of vector thrips, Frankliniella occidentalis, on symptomatic plants was also confirmed, suggesting a role in the spread of TSWV under greenhouse conditions. Recent surveys identified some greenhouses with 100% symptomatic peppers. The presence of TSWV in tomato and pepper has important implications for the domestic and export vegetable industry in the Dominican Republic because of the broad host range of the virus (4). It is critical for commercial producers to monitor TSWV and deploy appropriate management strategies to limit virus spread. References: (1) O. J. Alabi et al. J. Virol. Methods 154:111, 2008. (2) P.-G. S. Chang et al. J. Virol. Methods 171:345, 2011. (3) F. H. Chu et al. Phytopathology 91:361, 2001. (4) G. Parrella et al. J. Plant Pathol. 85:227, 2003.


An analysis of the functioning of mental healthcare in northwestern Poland.
Modern psychiatry faces numerous challenges related with the change of the epidemiology of mental disorders and the development of knowledge in this area of science. An answer to this situation is to be the introduction of community psychiatry. The implementation of this model in Poland was the aim of the National Mental Health Protection Programme. The aim of the study was to analyse the functioning of mental healthcare using the example of the West Pomeranian Province in Poland. The analysis relied on a qualitative method. Three group interviews in an interdisciplinary advisory panel were conducted. People representing various areas acting for people with mental disorders participated in each meeting. Based on the conclusions that were drawn, PEST and SWOT analyses of functioning of mental healthcare were performed. Within the analysis of the macro-environment of mental healthcare, the influence of the following factors was evaluated through PEST analysis: political and legal, economic, socio-cultural, and technological. All of these factors were assessed as negative for the functioning of mental healthcare. Then, a SWOT analysis was performed to indicate the strengths, weaknesses, opportunities, and threats in the functioning of mental healthcare. 1. Mental healthcare is more influenced by external factors than by internal factors. 2. Macro-environmental factors influence the functioning of mental healthcare in a significantly negative manner. 3. The basic problem in the functioning of mental healthcare is insufficient funding. 4. In order to improve the functioning of mental healthcare, it is necessary to change the funding methods, regulations, the way society perceives mental disorders, and the system of monitoring mental healthcare services.


Low-temperature anaerobic digestion is associated with differential methanogenic protein expression.
Anaerobic digestion (AD) is an attractive wastewater treatment technology, leading to the generation of recoverable biofuel (methane). Most industrial AD applications, carry excessive heating costs, however, as AD reactors are commonly operated at mesophilic temperatures while handling waste streams discharged at ambient or cold temperatures. Consequently, low-temperature AD represents a cost-effective strategy for wastewater treatment. The comparative investigation of key microbial groups underpinning laboratory-scale AD bioreactors operated at 37, 15 and 7°C was carried out. Community structure was monitored using 16S rRNA clone libraries, while abundance of the most prominent methanogens was investigated using qPCR. In addition, metaproteomics was employed to access the microbial functions carried out in situ. While δ-Proteobacteria were prevalent at 37°C, their abundance decreased dramatically at lower temperatures with inverse trends observed for Bacteroidetes and Firmicutes. Methanobacteriales and Methanosaeta were predominant at all temperatures investigated while Methanomicrobiales abundance increased at 15°C compared to 37 and 7°C. Changes in operating temperature resulted in the differential expression of proteins involved in methanogenesis, which was found to occur in all bioreactors, as corroborated by bioreactors' performance. This study demonstrated the value of employing a polyphasic approach to address microbial community dynamics and highlighted the functional redundancy of AD microbiomes.


Family response to the farm crisis: a study in coping.
The recent farm crisis created major economic disruptions for rural communities and families. To understand the effects of these disruptions on rural families, a study was conducted with community leaders and adult members of rural families that had experienced these disruptions. Family members experienced many stress reactions, especially withdrawal and depression. Families coped primarily by marshalling family efforts to solve financial problems and to provide emotional support. Significant barriers prevented people from turning to others in the extended family and the community for help. Family members created barriers by their withdrawal and their sense of individualism and shame. Judgmental attitudes and lack of understanding by others as well as demeaning and inappropriate criteria for help also created barriers. Interventions require monitoring policy issues and mobilizing community resources, as well as helping individual families to cope.


Immunomodulation and postpartum relapses in patients with multiple sclerosis.
Multiple sclerosis (MS) mainly affects young women during a life period with desire for children. Relapse rate decreases during pregnancy and rises after delivery. Therefore, studies on satisfactory postpartum relapse prevention and its efficacy are essential. Previous smaller and uncontrolled studies suggested that intravenous immunoglobulin (IVIG) administration reduced the relapse rate following delivery. The objective of our observational study was to compare the efficacy of IVIG application, treatment with other immunomodulatory compounds or no treatment at all on the postpartal relapse rate in female MS patients from our pregnancy database. One hundred and twenty four pregnancies were followed in a partly prospective design. Relapse rate was reduced during pregnancy (p50.001) and increased during the initial 3 months after delivery in all MS patients (p50.001). The relapse rate reduction showed only a trend in favour of the IVIG-treated women, probably due to the small number of patients. However, analysing the expected number of relapses, IVIG treated patients had significantly less relapses postpartum than the untreated control group matched for disease activity before and during pregnancy (χ(2), p= 0.013). The results suggest that IVIG could be an option to prevent postpartum relapse of MS.


Erythropoiesis-stimulating agents, hypertension and left ventricular hypertrophy in the chronic kidney disease patient.
Left-ventricular hypertrophy (LVH) represents an important marker of cardiovascular morbidity and mortality. Numerous noninterventional studies in patients with chronic kidney disease (CKD) revealed a consistent relationship of LVH with modifiable risk factors attributable to failing renal function, particularly anemia and hypertension. Given the clear role for anemia in initiating or accelerating LVH, it seems imperative to correct renal anemia with erythropoiesis-stimulating agents (ESAs). A few nonrandomized studies have described a regression of LVH with correction of anemia, but prospective randomized trials showed no evidence that ESA treatment is able to improve cardiac prognosis in the CKD patient. Current data alert physicians that normalization of hemoglobin in patients with advanced CKD is harmful. Recent studies are now trying to clarify the mechanisms for harm focussing on the influence of comorbidities, ESA doses, and hemoglobin variability. The pathogenesis of hypertension in CKD is multifactorial and only a small percentage of CKD patients have controlled their blood pressure, indicating poor medication adherence, insufficient control of volume overload and undertreatment. This review provides an update of ESA treatment, hypertension and LVH in the CKD patient, indicating that pathogenesis of LVH in this population is currently not completely understood. In addition, the impact of pharmacological interventions targeted to prevent or reduce LVH in anemic or hypertensive CKD patients is not well defined. As adoption of the Framingham approach seems not feasible in the CKD patient, evidence from large-scale randomized clinical trials is mandatory to resolve this dilemma.


Model-Based Feedforward Control of Part Height in Directed Energy Deposition.
Control of the geometric accuracy of a metal deposit is critical in the repair and fabrication of complex components through Directed Energy Deposition (DED). This paper developed and experimentally evaluated a model-based feedforward control of laser power with the objective of achieving the targeted part height in DED. Specifically, based on the dynamic model of melt-pool geometry derived from our prior work, a nonlinear inverse-dynamics controller was derived in a hatch-by-hatch, layer-by-layer manner to modulate the laser power such that the melt-pool height was regulated during the simulated build process. Then, the laser power trajectory from the simulated closed-loop control under the nonlinear inverse-dynamics controller was implemented as a feedforward control in an Optomec Laser-Engineered Net Shape (LENS) MR-7 system. This paper considered the deposition of L-shaped structures of Ti-6AL-4V as a case study to illustrate the proposed model-based controller. Experimental validation showed that by applying the proposed model-based feed-forward control for laser power, the resulting build had 24-42% reduction in the average build height error with respect to the target build height compared to applying a constant laser power through the entire build or applying a hatch-dependent laser power strategy, for which the laser power values were obtained from experimental trial and error.


Bradykinin directly triggers GLUT4 translocation via an insulin-independent pathway.
Physical exercise induces translocation of GLUT4 from an intracellular pool to the cell surface in skeletal muscles and increases glucose uptake via an insulin-independent pathway. However, the molecular mechanism remains to be identified. Some studies have suggested that bradykinin is locally released from contracting muscles and may be responsible for GLUT4 translocation and the increase of glucose transport in skeletal muscles. To determine whether bradykinin directly triggers GLUT4 translocation, we established L6 myotubes, 3T3-L1 adipocytes, and Chinese hamster ovary cells stably expressing c-myc epitope-tagged GLUT4 (GLUT4myc) and bradykinin B2 receptors. We found that bradykinin directly triggered GLUT4myc translocation and increased the rate of glucose uptake in a dose-dependent manner in these cells. The translocation with bradykinin occurred even after pretreatment with an islet-activating protein, wortmannin, and phorbol 12,13-dibutyrate. The signaling pathway does not seem to be mediated by Gi, phosphatidylinositol 3-kinase, or protein kinase C. It is insulin-independent and via trimeric G-protein Gq. Bradykinin is probably one of the factors responsible for exercise-stimulated glucose uptake in skeletal muscles.


Acute blood volume expansion delays the gastrointestinal transit of a charcoal meal in awake rats.
The present study evaluates the effect of blood volume expansion on the gastrointestinal transit of a charcoal meal (2.5 ml of an aqueous suspension consisting of 5% charcoal and 5% gum arabic) in awake male Wistar rats (200-270 g). On the day before the experiments, the rats were anesthetized with ether, submitted to left jugular vein cannulation and fasted with water ad libitum until 2 h before the gastrointestinal transit measurement. Blood volume expansion by i.v. infusion of 1 ml/min Ringer bicarbonate in volumes of 3, 4 or 5% body weight delayed gastrointestinal transit at 10 min after test meal administration by 21.3-26.7% (P < 0.05), but no effect was observed after 1 or 2% body weight expansion. The effect of blood volume expansion (up to 5% body weight) on gastrointestinal transit lasted for at least 60 min (P < 0.05). Mean arterial pressure increased transiently and central venous pressure increased and hematocrit decreased (P < 0.05). Subdiaphragmatic vagotomy and yohimbine (3 mg/kg) prevented the delay caused by expansion on gastrointestinal transit, while atropine (0.5 mg/kg), L-NAME (2 mg/kg), hexamethonium (10 mg/kg), prazosin (1 mg/kg) or propranolol (2 mg/kg) were ineffective. These data show that blood volume expansion delays the gastrointestinal transit of a charcoal meal and that vagal and yohimbine-sensitive pathways appear to be involved in this phenomenon. The delay in gastrointestinal transit observed here, taken together with the modifications of gastrointestinal permeability to salt and water reported by others, may be part of the mechanisms involved in liquid excess management.


A Novel Real-Time PCR Assay for the Rapid Detection of Virulent Streptococcus equi Subspecies zooepidemicus-An Emerging Pathogen of Swine.
Streptococcus equi subspecies zooepidemicus, a zoonotic bacterial pathogen caused a series of outbreaks with high mortality affecting swine herds in multiple locations of the USA and Canada in 2019. Further genetic analysis revealed that this agent clustered with ATCC 35246, a S. zooepidemicus strain associated with high mortality outbreaks in swine herds of China originally reported in 1977. Rapid and accurate diagnosis is absolutely critical for controlling and limiting further spread of this emerging disease of swine. Currently available diagnostic methods including bacteriological examination and PCR assays do not distinguish between the virulent strains and avirulent commensal strains of S. zooepidemicus, which is critical given that this pathogen is a normal inhabitant of the swine respiratory tract. Based on comparative analyses of whole genome sequences of the virulent isolates and avirulent sequences, we identified a region in the SzM gene that is highly conserved and restricted to virulent S. zooepidemicus strains. We developed and validated a novel probe-based real-time PCR targeting the conserved region of SzM. The assay was highly sensitive and specific to the virulent swine isolates of Streptococcus equi subspecies zooepidemicus. No cross reactivity was observed with avirulent S. zooepidemicus isolates as well as other streptococcal species and a panel of porcine respiratory bacterial and viral pathogens. The PCR efficiency of the assay was 96.64 % and was able to detect as little as 20 fg of the bacterial DNA. We then validated the diagnostic sensitivity and specificity of the new PCR assay using a panel of clinical samples (n = 57) and found that the assay has 100% sensitivity and specificity as compared to bacteriological culture method. In summary, the PCR assay will be an extremely valuable tool for the rapid accurate detection of virulent swine S. zooepidemicus isolates and directly from clinical samples.


Structure and meaning in models of breast and cervical cancer risk factors: a comparison of perceptions among Latinas, Anglo women, and physicians.
This article reports on a study of perceptions of breast and cervical cancer risk factors among 27 U.S.-born Chicanas, 39 Mexican and 28 Salvadoran immigrants, 27 Anglo women, and 30 physicians in northern Orange County, California. In open-ended responses explaining why women might be at risk for both cancers, Latinas expressed two general themes: physical stress and trauma to the body, and behavior and lifestyle choices. Interviewees ranked the specific risk factors that they themselves mentioned. Cultural consensus of ranked data revealed that Mexican and Salvadoran immigrants had a model of cancer risks that was different from those of Anglo women and physicians. U.S.-born Chicanas were bicultural in their views, which overlapped with both Mexican women's and Anglo women's views, but less so with physicians' views. Comparing views about the two cancers revealed that general themes apply across both cancers, that Latina immigrants agreed less on the risk factors for cervical cancer than for breast cancer, and that there is a consistent pattern in the different ways Latinas, Anglos, and physicians perceive risk factors for both cancers.


Influenza virus genotype to phenotype predictions through machine learning: a systematic review.
There is great interest in understanding the viral genomic predictors of phenotypic traits that allow influenza A viruses to adapt to or become more virulent in different hosts. Machine learning techniques have demonstrated promise in addressing this critical need for other pathogens because the underlying algorithms are especially well equipped to uncover complex patterns in large datasets and produce generalizable predictions for new data. As the body of research where these techniques are applied for influenza A virus phenotype prediction continues to grow, it is useful to consider the strengths and weaknesses of these approaches to understand what has prevented these models from seeing widespread use by surveillance laboratories and to identify gaps that are underexplored with this technology. We present a systematic review of English literature published through 15 April 2021 of studies employing machine learning methods to generate predictions of influenza A virus phenotypes from genomic or proteomic input. Forty-nine studies were included in this review, spanning the topics of host discrimination, human adaptability, subtype and clade assignment, pandemic lineage assignment, characteristics of infection, and antiviral drug resistance. Our findings suggest that biases in model design and a dearth of wet laboratory follow-up may explain why these models often go underused. We, therefore, offer guidance to overcome these limitations, aid in improving predictive models of previously studied influenza A virus phenotypes, and extend those models to unexplored phenotypes in the ultimate pursuit of tools to enable the characterization of virus isolates across surveillance laboratories.


Liver cell dysplasia in cirrhosis. A serologic and immunohistochemical study.
Liver cell dysplasia (LCD) was investigated for hepatitis B virus (HBV) markers, alpha-fetoprotein (AFP) and ferritin by serologic and immunohistochemical methods in 101 patients with cirrhosis. LCD was found in 30 cases (29.7%), with the highest incidence in cases of posthepatitic cirrhosis (67%). In the group of dysplastic cirrhosis (DC) 46.6% of the patients had active HBV infection (hepatitis B surface antigen [HBsAg] serum positivity) compared with 7% of the patients with nondysplastic cirrhosis (NDC) (P less than 0.01). The mean serum AFP concentration was significantly raised in the DC group compared with that in the NDC group (P less than 0.05). In seven patients with LCD at the initial biopsy, the histologic followup showed the persistence of LCD in all cases, and the development of hepatocellular carcinoma (HCC) in three cases. In serologic HBsAg-positive cases, dysplastic cells, at variance with the surrounding liver parenchyma, were almost always negative for tissue HBsAg, and always negative for tissue hepatitis B core antigens (HBcAg). AFP was never detected in either normal or dysplastic cells. Ferritin was found in all cases, but dysplastic foci displayed a lesser amount of this protein. These serologic and immunohistochemical data strongly suggest a preneoplastic significance of LCD. The importance of monitoring cirrhotic patients with LCD and particularly those with HBV infection and/or increased AFP levels with more aggressive follow-up is also stressed.


Oligonucleotide fishing for STAT6: cross-talk between IL-4 and chemokines.
Signal transducer and activator of transcription 6 (STAT6) is essential for the biological activities of interleukin-4 (IL-4) and the development of allergic responses in mice. Here we report on a sensitive and specific assay for STAT6 activation in response to IL-4. We took advantage of double-stranded oligonucleotide probes containing a STAT6-binding gene-sequence from the promotor of the immunoglobulin heavy chain germline epsilon transcript to study the IL-4-induced DNA binding of STAT6. Using these probes, we show that repeated adjacent STAT6-binding sites result in enhanced STAT6-DNA binding. Moreover, the distance between the binding sites is critical for STAT-DNA binding, i.e. STAT6 binding is decreased at distances above 20 nucleotides between neighbouring binding sites. Using this assay to study cross-talk between IL-4 and chemokines, we provide evidence that MIP-1beta and MIG inhibit IL-4-induced STAT6 activation, whereas other chemokines and cytokines do not. In conclusion, our data show that oligonucleotide fishing is a supplementary tool for studying cytokine cross-talk at a genomic level.


Investigation of the first step of biotin biosynthesis in Bacillus sphaericus. Purification and characterization of the pimeloyl-CoA synthase, and uptake of pimelate.
The pimeloyl-CoA synthase from Bacillus sphaericus has been purified to homogeneity from an overproducing strain of Escherichia coli. The purification yielded milligram quantities of the synthase with a specific activity of 1 unit/mg of protein. Analysis of the products showed that this enzyme catalysed the transformation of pimelate into pimeloyl-CoA with concomitant hydrolysis of ATP to AMP. Using a continuous spectrophotometric assay, we have examined the catalytic properties of the pure enzyme. The pH profile under Vmax. conditions showed a maximum around 8.5. Apparent Km values for pimelate, CoASH, ATP.Mg2- and Mg2+ were respectively 145 microM, 33 microM, 170 microM and 2.3 mM. The enzyme was inhibited by Mg2+ above 10 mM. This acid-CoA ligase exhibited a very sharp substrate specificity, e.g. neither GTP nor pimelate analogues (di- or mono-carboxylic acids) were processed. The bivalent metal ion requirement was also investigated: Mn2+ (73%) and Co2+ (32%) but not Ca2+ could replace Mg2+. The enzyme was inhibited by metal chelators such as 1,10-phenanthroline and EDTA. The synthase was a homodimer with a 28,000-M(r) subunit. N-Terminal sequencing definitely proved that this enzyme was encoded by the bioW gene. A careful study of pimelate uptake by B. sphaericus, E. coli and Pseudomonas dentrificans showed that this metabolite crossed the membrane of these microorganisms by passive diffusion, ruling out the involvement of the bioX gene product as pimelate carrier.


DEVELOPMENT OF THE ONTARIO DECISION FRAMEWORK: A VALUES BASED FRAMEWORK FOR HEALTH TECHNOLOGY ASSESSMENT.
In 2007, the Ontario Health Technology Advisory Committee (OHTAC) developed a decision framework to guide decision making around nondrug health technologies. In 2012, OHTAC commissioned a revision of this framework to enhance its usability and deepen its conceptual and theoretical foundations. The committee overseeing this work used several methods: (a) a priori consensus on guiding principles, (b) a scoping review of decision attributes and processes used globally in health technology assessment (HTA), (c) presentations by methods experts and members of review committees, and (d) committee deliberations over a period of 3 years. The committee adopted a multi-criteria decision-making approach, but rejected the formal use of multi-criteria decision analysis. Three broad categories of attributes were identified: (I) context criteria attributes included factors such as stakeholders, adoption pressures from neighboring jurisdictions, and potential conflicts of interest; (II) primary appraisal criteria attributes included (i) benefits and harms, (ii) economics, and (iii) patient-centered care; (III) feasibility criteria attributes included budget impact and organizational feasibility. The revised Ontario Decision Framework is similar in some respects to frameworks used in HTA worldwide. Its distinctive characteristics are that: it is based on an explicit set of social values; HTA paradigms (evidence based medicine, economics, and bioethics/social science) are used to aggregate decision attributes; and that it is rooted in a theoretical framework of optimal decision making, rather than one related to broad social goals, such as health or welfare maximization.


Field theoretical analysis of driving forces for the uptake of proteins by like-charged polyelectrolyte brushes: effects of charge regulation and patchiness.
At the moment two competing explanations exist for the experimental finding that net negatively charged proteins adsorb on or absorb in negatively charged polyelectrolyte brushes. One explanation is based on the possibility of charge regulation. The idea is that a protein can reverse its charge when it is in the presence of the high electrostatic potential of the brush and then can be inserted. The other explanation relies on the charge anisotropy of proteins, that is, that it carries positively charged and negatively charged patches. The positively charged region gains more energy from interacting with the negative brush than the negative charged patch loses, especially when the charge densities and electrostatic potentials are high, thus providing a net attraction. We present a model in which both mechanisms are combined. We confirm that both charge anisotropy and charge regulation effects on their own can be responsible for protein uptake at the "wrong" side of the isoelectric point (IEP). In addition, we find that the respective effects are additive. Indeed, taking both effects into account results in a stronger attraction between a PE brush and protein at the IEP, and the attraction is found further above the IEP than the individual effects would have made possible. Still, for patchiness to have a strong contribution, the patches need very high charge densities. Therefore, we argue that for most types of protein charge reversal will be the main driving force for adsorption on the wrong side of the IEP, while patchiness will contribute less.


[Ultrasonography-guided percutaneous drainage of hepatic abscess. Results and complications].
Interventional echography has been proposed as an effective method for the drainage of pyogenic hepatic abscesses. We present the technique and the results with this therapeutic alternative in 23 patients suffering from this serious disease. The procedure was effective in 19 patients (82.6%), being palliative in 3 cases (a carcinoma of the gallbladder, a biliary-bowel fistula and a gastric perforation). The drainage failed in 4 patients (17.4%) (in an infected hematoma were and in 3 cases due to technical problems). There were 4 complications, one of them severe (pleural empyema). We consider that percutaneous drainage guided by echography should by a first choice in the treatment of hepatic abscesses. To achieve good results and to avoid complications it is necessary to have good knowledge of the technique.


[Acute appendicitis. Preoperative observation time and diagnostic accuracy].
The indication for operative treatment of suspected acute appendicitis is based mainly on clinical examination. In this retrospective study of 327 emergency laparotomies the accuracy of the surgeons diagnosis was 69.4%. The overall rate of unnecessary laparotomy was 28.4%, and 45.6% of the operated females, aged 12 to 40 years, did not actually require surgery. The median postadmission delay before operation was three hours 28 minutes. With increasing observation time, the diagnostic accuracy was significantly decreased from 77.7 to 52.2%, and the incidence of perforation rose from 18.6 to 33.3%. 8.7% of the patients with a surgical condition suffered from excessive delay (greater than 12 hours) before surgery. There was no correlation between time of admission and operation and accuracy of diagnosis, but a considerable variation in accuracy between different surgeons. The difficulty in obtaining a correct pre-operative diagnosis in patients with suspected acute appendicitis is still a clinical problem of major concern.


Use of frameless stereotactic computed tomography venography for intraoperative localization of dural arterial venous fistulas: case report.
The following technical case report illustrates the use of multiple imaging modalities including 3D CTV with frameless stereotactic navigation for intraoperative planning and localization for surgical obliteration of a cranial dAVF with leptomeningeal drainage. This 65-year-old man presented with an asymptomatic occipital dAVF with leptomeningeal drainage. In addition to cerebral angiography, a CTV with 3D reconstruction was performed, which provided excellent visualization of the dAVF and clarified its pattern of drainage. The dAVF was supplied by a middle meningeal artery branch that drained into an occipital cortical vein, which then retrograde filled the vein of Labbé. Frameless stereotactic navigation with the imported CTV images was used to plan the craniotomy and to localize the leptomeningeal draining vein and vein of Labbé. The draining vein of the fistula was successfully ligated and divided while preserving flow in the vein of Labbé. Postoperative angiogram demonstrated complete obliteration of the dAVF. Integration of a 3D reconstructed CTV with conventional angiographic information optimized our surgical understanding of the spatial anatomy of this dAVF and its pattern of venous drainage. Applying the CTV with frameless stereotaxy allowed for safe obliteration of the dAVF while preserving the vein of Labbé.


Oxygen-evolving extrinsic proteins (PsbO,P,Q,R): bioinformatic and functional analysis.
The water-splitting and oxygen-evolving (OE) reaction is carried out by a large multisubunit protein complex, Photosystem II (PSII), that has two distinct regions: a membrane intrinsic-region that includes most of the PSII subunits and a lumenal extrinsic-region that is in close association to the manganese catalytic center. The recently determined PSII 3D structures from cyanobacteria provide a considerable amount of new knowledge about the OE architecture (K.N. Ferreira, T.M. Iverson, K. Maghlaoui, J. Barber, S. Iwata, Architecture of the photosynthetic oxygen-evolving center, Science 303 (2004) 1831-1838; B. Loll, J. Kern, W. Saenger, A. Zouni, J. Biesiadka, Towards complete cofactor arrangement in the 3.0 A resolution structure of photosystem II, Nature 438 (2005) 1040-1044). Most of the intrinsic core PSII polypeptides have been well conserved through evolution from ancient cyanobacteria to modern plants, keeping the essence of PSII light driven reactions from prokaryotes to eukaryotes; but what is striking is the large number of changes that have occurred in the oxygen-evolving extrinsic proteins (OEEp) associated to PSII lumenal side. For unknown reasons plant PSII has required the "invention" of three OEEps: PsbP (23 kDa), PsbQ (16 kDa) and PsbR (10 kDa); associated to the ubiquitous OEEp PsbO (33 kDa). This set of proteins seems to be required in plants for the full activity and stability of the OE center in vivo, but their specific function is not clear. In this paper, bioinformatics and functional data show that the OEEps present in plants and green algae are very distinct from their prokaryotic counterparts. Moreover, clear differences are found for PsbQ from higher plants and green algae; and a relationship has been found between PsbR and the Mn cluster.


Adalimumab in Patients with Active Noninfectious Uveitis.
Patients with noninfectious uveitis are at risk for long-term complications of uncontrolled inflammation, as well as for the adverse effects of long-term glucocorticoid therapy. We conducted a trial to assess the efficacy and safety of adalimumab as a glucocorticoid-sparing agent for the treatment of noninfectious uveitis. This multinational phase 3 trial involved adults who had active noninfectious intermediate uveitis, posterior uveitis, or panuveitis despite having received prednisone treatment for 2 or more weeks. Investigators and patients were unaware of the study-group assignments. Patients were randomly assigned in a 1:1 ratio to receive adalimumab (a loading dose of 80 mg followed by a dose of 40 mg every 2 weeks) or matched placebo. All patients received a mandatory prednisone burst followed by tapering of prednisone over the course of 15 weeks. The primary efficacy end point was the time to treatment failure occurring at or after week 6. Treatment failure was a multicomponent outcome that was based on assessment of new inflammatory lesions, best corrected visual acuity, anterior chamber cell grade, and vitreous haze grade. Nine ranked secondary efficacy end points were assessed, and adverse events were reported. The median time to treatment failure was 24 weeks in the adalimumab group and 13 weeks in the placebo group. Among the 217 patients in the intention-to-treat population, those receiving adalimumab were less likely than those in the placebo group to have treatment failure (hazard ratio, 0.50; 95% confidence interval, 0.36 to 0.70; P<0.001). Outcomes with regard to three secondary end points (change in anterior chamber cell grade, change in vitreous haze grade, and change in best corrected visual acuity) were significantly better in the adalimumab group than in the placebo group. Adverse events and serious adverse events were reported more frequently among patients who received adalimumab (1052.4 vs. 971.7 adverse events and 28.8 vs. 13.6 serious adverse events per 100 person-years). In our trial, adalimumab was found to be associated with a lower risk of uveitic flare or visual impairment and with more adverse events and serious adverse events than was placebo. (Funded by AbbVie; VISUAL I ClinicalTrials.gov number, NCT01138657 .).


Curtailing laboratory test ordering in a managed care setting through redesign of a computerized order form.
To increase appropriate use of blood tests for folic acid and vitamin B12 ordered by primary care physicians in a managed care organization in Israel through redesign of a computerized order form. Pre-post intervention. A new version of the computerized order form was launched. Utilization patterns were calculated for tests of vitamin B12, folic acid, and ferritin, which were previously grouped together. Concomitant utilization patterns for tests of hemoglobin and iron were evaluated as controls. Tests ordered for the 3 targets decreased by 31% to 41% relative to the preintervention month, with a further decrease to 36% to 53% the following month. Negligible changes in utilization patterns were observed for the controls (-2% to 3%) during the postintervention period. Simple restructuring of a computerized order form significantly reduced the number of laboratory tests suspected of being unnecessary or redundant. When overutilization of laboratory resources is suspected, managers should evaluate the efficiency of the organization's current ordering procedures before implementing resource-intensive interventions.


Different sites of photodamage in chilling-sensitive (sorghum) and chilling-resistant (barley and wheat) plants.
The electron transport chain was affected to varying degrees by high light intensity at low temperature in different crop plants. The PS II was found to be the sensitive site while PS I showed very little change in its activity. Photoinhibition affected the oxidizing side of PS II in all three plants. However, the site of inhibition was different in chilling-sensitive and chilling-resistant plants. In sorghum, the water splitting reaction was damaged while in barley and wheat the damage occurred in the reaction centre itself. It appears that photoinhibition may affect different sites within the PS II in chilling-resistant and sensitive plants.


Radiographic Measurements of the Affected and Unaffected Feet in Patients with Unilateral Hallux Limitus A Case-Control Pilot Study.
Controversy exists regarding the structural and functional causes of hallux limitus, including metatarsus primus elevatus, a long first metatarsal, first-ray hypermobility, the shape of the first metatarsal head, and the presence of hallux interphalangeus. Some articles have reported on the radiographic evaluation of these measurements in feet affected by hallux limitus, but no study has directly compared the affected and unaffected feet in patients with unilateral hallux limitus. This case-control pilot study aimed to establish whether any such differences exist. Dorsoplantar and lateral weightbearing radiographs of both feet in 30 patients with unilateral hallux limitus were assessed for grade of disease, lateral intermetatarsal angle, metatarsal protrusion distance, plantar gapping at the first metatarsocuneiform joint, metatarsal head shape, and hallux abductus interphalangeus angle. Data analysis was performed using a statistical software program. Mean radiographic measurements for affected and unaffected feet demonstrated that metatarsus primus elevatus, a short first metatarsal, first-ray hypermobility, a flat metatarsal head shape, and hallux interphalangeus were prevalent in both feet. There was no statistically significant difference between feet for any of the radiographic parameters measured (Mann-Whitney U tests, independent-samples t tests, and Pearson χ(2) tests: P > .05). No significant differences exist in the presence of the structural risk factors examined between affected and unaffected feet in patients with unilateral hallux limitus. The influence of other intrinsic factors, including footedness and family history, should be investigated further.


Midterm stability and hemodynamic performance of a transfemorally implantable nonmetallic, retrievable, and repositionable aortic valve in patients with severe aortic stenosis. Up to 2-year follow-up of the direct-flow medical valve: a pilot study.
Misplacement during percutaneous aortic valve implantation can be associated with severe complications. The direct flow medical (DFM) valve is repositionable and retrievable; however, the nonmetallic inflatable and conformable design of the valve results in less radial force, which may have an impact on stability and valve function over time. We, therefore, analyzed the midterm stability of the position, shape, and hemodynamic performance of the DFM percutaneous aortic valve. Sixteen symptomatic high-risk for surgery patients with aortic stenosis and a logistic EuroSCORE >20 underwent implantation and were the subject of this analysis. Clinical, echocardiographic, and dual-source multislice computed tomography data were obtained during 2-year follow-up. The 1- and 2-year survival rates were 81% and 69%, respectively. The dual-source multislice computed tomography follow-up indicated no changes in position, diameter, and orifice area of the DFM valve over time. Echocardiography revealed a significant decrease of the mean gradient from baseline (50.1±11.3 mm Hg) to 30 days (19.6±5.7 mm Hg, P<0.001), which remained stable over 2 years. The aortic valve area increased from 0.57±0.15 cm(2) at baseline to 1.47±0.35 cm(2) at 30 days (P<0.001) and did not significantly change during 2-year follow-up. Of the patients, 73% had no aortic regurgitation (AR) and 27% had minimal AR. In this preliminary series, the 2-year follow-up data of patients, in whom the nonmetallic, repositionable, and retrievable DFM valve was successfully implanted, show stability of the position, shape, and hemodynamic performance, with no AR in most patients.


[Primary carcinoma of the male prostatic urethra. Report of a case and review of the literature].
A case of advanced primary transitional cell carcinoma of the prostatic urethra (stage D1 of the Ray classification) is described. The patient was submitted to TUR and received a radical dose of external radiotherapy. Patient follow up at 22 months has revealed no local recurrence or distant metastasis. The literature is reviewed with a special reference to the therapeutic approach.


Harnessing host-virus evolution in antiviral therapy and immunotherapy.
Pathogen resistance and development costs are major challenges in current approaches to antiviral therapy. The high error rate of RNA synthesis and reverse-transcription confers genome plasticity, enabling the remarkable adaptability of RNA viruses to antiviral intervention. However, this property is coupled to fundamental constraints including limits on the size of information available to manipulate complex hosts into supporting viral replication. Accordingly, RNA viruses employ various means to extract maximum utility from their informationally limited genomes that, correspondingly, may be leveraged for effective host-oriented therapies. Host-oriented approaches are becoming increasingly feasible because of increased availability of bioactive compounds and recent advances in immunotherapy and precision medicine, particularly genome editing, targeted delivery methods and RNAi. In turn, one driving force behind these innovations is the increasingly detailed understanding of evolutionarily diverse host-virus interactions, which is the key concern of an emerging field, neo-virology. This review examines biotechnological solutions to disease and other sustainability issues of our time that leverage the properties of RNA and DNA viruses as developed through co-evolution with their hosts.


Influence of structural modifications at the 3' and 4' positions of doxorubicin on the drug ability to trap topoisomerase II and to overcome multidrug resistance.
To better define the role of the amino sugar in the pharmacological and biochemical properties of anthracyclines related to doxorubicin and daunorubicin, we have investigated the effects of various substituents at the 3'- and 4'-positions of the drug on cytotoxic activity and ability to stimulate DNA cleavage mediated by DNA topoisomerase II. The study shows that the nature of the substituent at the 3'-position but not the 4'-position is critical for drug ability to form cleavable complexes. The amino group at the 3'-position is not essential for cytotoxic and topoisomerase II-targeting activities, because it can be replaced by a hydroxyl group without reduction of activity. However, the presence of bulky substituents at this position (i.e., morpholinyl derivatives) totally inhibited the effects on the enzyme, thus supporting previous observations indicating that the cytotoxic potencies of these particular derivatives are not related to topoisomerase II inhibition. This conclusion is also supported by the observation that 3'-morpholinyl and 3'-methoxymorpholinyl derivatives are able to overcome atypical (i.e., topoisomerase II-mediated) multidrug resistance. Because a bulky substituent at the 4'-position did not reduce the ability to stimulate DNA cleavage, these results support a critical role of the 3'-position in the drug interaction with topoisomerase II in the ternary complex. An analysis of patterns of cross-resistance to the studied derivatives in resistant human tumor cell lines expressing different resistance mechanisms indicated that chemical modifications at the 3'-position of the sugar may have a relevant influence on the ability of the drugs to overcome specific mechanisms of resistance.


TYPE-SPECIFIC PROTECTION AND IMMUNITY FOLLOWING INTRANASAL INOCULATION OF MONKEYS WITH GROUP A HEMOLYTIC STREPTOCOCCI.
1. Nasopharyngeal carrier states of several weeks to several months duration were induced in the Macaca mulatta by the intranasal inoculation with matt strains of group A streptococci. 2. Following such a successful inoculation with a particular type of group A streptococcus, the animal was usually resistant to reimplantation with that same type for several months to a year or more, although reimplantation with a heterologous type could generally be easily effected. 3. This resistance was shown to be closely correlated with the antibodies directed toward the type-specific M antigen, not toward the T antigen of the strains employed. 4. A majority (83.8 per cent) of the animals in which intranasal inoculation was followed by successful implantation developed significant increases in the antistreptolysin O titres of their sera; and in a limited number of instances, type-specific agglutinins and bacteriostatic antibodies were demonstrable in the animals' sera following successful implantation. 5. Nasopharyngeal carrier states could not be induced with the glossy, avirulent variants of group A streptococci; these animals, moreover, failed to show antibody responses and were susceptible to implantation with the matt variants of the homologous glossy strains. 6. The findings are in accord with the known facts regarding immunity to group A streptococci gained through experiments on rodents; and the possible relationship of these observations to the problem of type-specific immunity in human beings is discussed.


[Diamond-Blackfan anemia with extremely low hemoglobin].
Diamond-Blackfan anaemia (DBA) has been described in approximately 300 cases world wide. It is characterised by a normochromic-macrocytic anaemia in early childhood, reticulocytopenia and a normocellular marrow with a selective deficiency of red cell precursors. Foetal haemoglobin is usually increased, and DBA is associated with increased expression of "i" antigen. DBA is often associated with multiple congenital anomalies, which are described. Transient erythroblastopenia is an important differential diagnosis and the distinctive characteristics are discussed. A case of DBA in an 8-week-old boy presenting with extremely low haemoglobin (0.9 mmol/l) is presented. The literature is reviewed.


Localization of endogenous and recombinant Na(+)-driven anion exchanger protein NDAE1 from Drosophila melanogaster.
Na(+)-dependent Cl(-)/HCO exchange activity helps maintain intracellular pH (pH(i)) homeostasis in many invertebrate and vertebrate cell types. Our laboratory cloned and characterized a Na(+)-dependent Cl(-)/HCO exchanger (NDAE1) from Drosophila melanogaster (Romero MF, Henry D, Nelson S, Harte PJ, and Sciortino CM. J Biol Chem 275: 24552--24559, 2000). In the present study we used immunohistochemical and Western blot techniques to characterize the developmental expression, subcellular localization, and tissue distribution of NDAE1 protein in D. melanogaster. We have shown that a polyclonal antibody raised against the NH(2) terminus of NDAE1 (alpha CWR57) recognizes NDAE1 electrophysiologically characterized in Xenopus oocytes. Moreover, our results begin to delineate the NDAE1 topology, i.e., both the NH(2) and COOH termini are intracellular. NDAE1 is expressed throughout Drosophila development in the central and peripheral nervous systems, sensilla, and the alimentary tract (Malpighian tubules, gut, and salivary glands). Coimmunolabeling of larval tissues with NDAE1 antibody and a monoclonal antibody to the Na(+)-K(+)-ATPase alpha-subunit revealed that the majority of NDAE1 is located at the basolateral membranes of Malpighian tubule cells. These results suggest that NDAE1 may be a key pH(i) regulatory protein and may contribute to basolateral ion transport in epithelia and nervous system of Drosophila.


Release of dopamine from coaggregate cultures of mesencephalic tegmentum and corpus striatum.
Dissociated dopamine (DA) neurons from 14-day fetal mice were dissected from the rostral mesencephalic tegmentum (RMT) and were allowed to reaggregate in vitro with cells from the corpus striatum (CS). As previously demonstrated under these conditions, DA neurons develop punctate fluorescent varicosities and the capacity to synthesize, accumulate, and retain DA (Kotake, C., P. C. Hoffmann, and A. Heller (1982) J. Neurosci. 2: 1307-1315). After 17 to 22 days in culture, the RMT-CS coaggregates were assessed for their ability to release DA. Coaggregates were incubated in 5.6 x 10(-6) M [3H]DA, washed, and then superfused at 100 microliters/min for 2 hr. Fractions were collected every 2 min. Basal efflux of [3H]DA/2 min was 1% of tissue stores of 3H. K+, 70 mM infused for 8 min induced a peak release of 5.87% of tissue stores of 3H, and 50 mM K+ induced a peak release of 2.13%. The potassium-induced release of [3H]DA was calcium dependent. When d-amphetamine was infused for 12 min, 100 microM solutions induced a peak release of 8.91%, 10 microM induced a peak release of 4.36%, and 1 microM induced a peak release of 1.85% of tissue stores of 3H. Substance P at 100 microM induced a peak release of 2.19% of tissue stores of 3H. Tetrodotoxin (0.5 and 2.5 microM) decreased basal efflux by 40%, blocked substance P-induced release, but did not affect either potassium- or d-amphetamine-induced release of [3H]dopamine.


Decreasing Poly(ADP-Ribose) Polymerase Activity Restores ΔF508 CFTR Trafficking.
Most cystic fibrosis is caused by mutations in CFTR that prevent its trafficking from the ER to the plasma membrane and is associated with exaggerated inflammation, altered metabolism, and diminished responses to oxidative stress. PARP-1 is activated by oxidative stress and causes energy depletion and cell dysfunction. Inhibition of this enzyme protects against excessive inflammation and recent studies have also implicated it in intracellular protein trafficking. We hypothesized that PARP-1 activity is altered in CF and affects trafficking and function of the most common CF mutant ΔF508 CFTR. Indeed, PARP-1 activity was 2.9-fold higher in CF (ΔF508/ΔF508) human bronchial epithelial primary cells than in non-CF cells, and similar results were obtained by comparing CF vs. non-CF bronchial epithelial cell lines (2.5-fold higher in CFBE41o(-) vs. 16HBE14o(-), P < 0.002). A PARP-1 inhibitor (ABT-888, Veliparib) partially restored CFTR channel activity in CFBE41o(-) cells overexpressing ΔF508 CFTR. Similarly, reducing PARP-1 activity by 85% in ileum from transgenic CF mice (Cftr(tm1)Eur) partially rescued ΔF508 CFTR activity to 7% of wild type mouse levels, and similar correction (7.8%) was observed in vivo by measuring salivary secretion. Inhibiting PARP-1 with ABT-888 or siRNA partially restored ΔF508 CFTR trafficking in cell lines, and most ΔF508 CFTR was complex glycosylated when heterologously expressed in PARP-1(-/-) mouse embryonic fibroblasts. Finally, levels of the mature glycoform of CFTR were reduced by peroxynitrite, a strong activator of PARP-1. These results demonstrate that PARP-1 activity is increased in CF, and identify a novel pathway that could be targeted by proteostatic correctors of CFTR trafficking.


Omega-3 Fatty acids: anti-arrhythmic, pro-arrhythmic, or both?
This review focuses on developments after 2008, when the topic was last reviewed by the author. Pertinent publications were found by medline searches and in the author's personal data base. Prevention of atrial fibrillation (AF) was investigated in a number of trials, sparked by one positive report on the effects of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), considerations of upstream therapy, data from electrophysiologic laboratories and animal experiments. If EPA + DHA prevent postoperative AF, the effect is probably smaller than initially expected. The same is probably true for maintenance of sinus rhythm after cardioversion and for new-onset AF. Larger trials are currently ongoing. Prevention of ventricular arrhythmias was studied in carriers of an implanted cardioverter-defibrillator, with no clear results. This might have been due to a broad definition of the primary endpoint, including any ventricular arrhythmia and any action of the device. Epidemiologic studies support the contention that high levels of EPA + DHA prevent sudden cardiac death (SCD). However, since SCD is a rare occurrence, it is difficult to conduct an adequately powered trial. In patients with congestive heart failure, EPA + DHA reduced total mortality and rehospitalizations, but not SCD or presumed arrhythmic death. Of three trials in patients after a myocardial infarction, two were inadequately powered, and in one, the dose might have been too low. Taken together, while epidemiologic studies support an inverse relation between EPA + DHA and occurrence of SCD or arrhythmic death, demonstrating this effect in intervention trials remained elusive so far. A pro-arrhythmic effect of EPA + DHA has not been seen in intervention studies, and results of epidemiologic and animal studies also rather argue against such an effect. A different, and probably more productive, perspective is provided by a standardized analytical assessment of a person's status in EPA + DHA by use of the omega-3 index, EPA + DHA in red cell fatty acids. In populations with a high omega-3 index, SCD is rare. Intervention trials can become more effective by including a low omega-3 index into the inclusion criteria, thus creating a study population more likely to demonstrate an effect of EPA + DHA. This is especially relevant in case of rare endpoints, like new-onset AF or SCD.


Genu varum and genu valgum in children.
Genu varum and genu valgum are often normal developmental changes in knee alignment that occur in a young child. Measuring the child's height and understanding normal development is the key to determining which children need further evaluation for a possible skeletal or metabolic bone dysplasia.


Predicting cardiovascular risk using conventional vs ambulatory blood pressure in older patients with systolic hypertension. Systolic Hypertension in Europe Trial Investigators.
The clinical use of ambulatory blood pressure (BP) monitoring requires further validation in prospective outcome studies. To compare the prognostic significance of conventional and ambulatory BP measurement in older patients with isolated systolic hypertension. Substudy to the double-blind placebo-controlled Systolic Hypertension in Europe (Syst-Eur) Trial, started in October 1988 with follow up to February 1999. The conventional BP at randomization was the mean of 6 readings (2 measurements in the sitting position at 3 visits 1 month apart). The baseline ambulatory BP was recorded with a noninvasive intermittent technique. Family practices and outpatient clinics at primary and secondary referral hospitals. A total of 808 older (aged > or =60 years) patients whose untreated BP level on conventional measurement at baseline was 160 to 219 mm Hg systolic and less than 95 mm Hg diastolic. For the overall study, patients were randomized to nitrendipine (n = 415; 10-40 mg/d) with the possible addition of enalapril (5-20 mg/d) and/or hydrochlorothiazide (12.5-25.0 mg/d) or to matching placebos (n = 393). Total and cardiovascular mortality, all cardiovascular end points, fatal and nonfatal stroke, and fatal and nonfatal cardiac end points. After adjusting for sex, age, previous cardiovascular complications, smoking, and residence in western Europe, a 10-mm Hg higher conventional systolic BP at randomization was not associated with a worse prognosis, whereas in the placebo group, a 10-mm Hg higher 24-hour BP was associated with an increased relative hazard rate (HR) of most outcome measures (eg, HR, 1.23 [95% confidence interval [CI], 1.00-1.50] for total mortality and 1.34 [95% CI, 1.03-1.75] for cardiovascular mortality). In the placebo group, the nighttime systolic BP (12 AM-6 AM) more accurately predicted end points than the daytime level. Cardiovascular risk increased with a higher night-to-day ratio of systolic BP independent of the 24-hour BP (10% increase in night-to-day ratio; HR for all cardiovascular end points, 1.41; 95% CI, 1.03-1.94). At randomization, the cardiovascular risk conferred by a conventional systolic BP of 160 mm Hg was similar to that associated with a 24-hour daytime or nighttime systolic BP of 142 mm Hg (95% CI, 128-156 mm Hg), 145 mm Hg (95% CI, 126-164 mm Hg) or 132 mm Hg (95% CI, 120-145 mm Hg), respectively. In the active treatment group, systolic BP at randomization did not significantly predict cardiovascular risk, regardless of the technique of BP measurement. In untreated older patients with isolated systolic hypertension, ambulatory systolic BP was a significant predictor of cardiovascular risk over and above conventional BP.


A non-canonical function of Plk4 in centriolar satellite integrity and ciliogenesis through PCM1 phosphorylation.
Centrioles are the major constituents of the animal centrosome, in which Plk4 kinase serves as a master regulator of the duplication cycle. Many eukaryotes also contain numerous peripheral particles known as centriolar satellites. While centriolar satellites aid centriole assembly and primary cilium formation, it is unknown whether Plk4 plays any regulatory roles in centriolar satellite integrity. Here we show that Plk4 is a critical determinant of centriolar satellite organisation. Plk4 depletion leads to the dispersion of centriolar satellites and perturbed ciliogenesis. Plk4 interacts with the satellite component PCM1, and its kinase activity is required for phosphorylation of the conserved S372. The nonphosphorylatable PCM1 mutant recapitulates phenotypes of Plk4 depletion, while the phosphomimetic mutant partially rescues the dispersed centriolar satellite patterns and ciliogenesis in cells depleted of PCM1. We show that S372 phosphorylation occurs during the G1 phase of the cell cycle and is important for PCM1 dimerisation and interaction with other satellite components. Our findings reveal that Plk4 is required for centriolar satellite function, which may underlie the ciliogenesis defects caused by Plk4 dysfunction.


Verification of a standardized method for inserting intramuscular EMG electrodes into uniquely oriented segments of gluteus minimus and gluteus medius.
Guidelines for assessing the function of gluteus minimus and gluteus medius with electromyography (EMG) traditionally offer one electrode placement site per muscle. However, anatomical studies suggest that there are two uniquely oriented segments within gluteus minimus (anterior and posterior), and three within gluteus medius (anterior, middle, and posterior) with potential for independent function. Assessment of these muscles with one electrode may therefore provide only a limited account of their role. Thus, the aim of this cadaveric study was to verify guidelines for placing intramuscular electrodes into two uniquely oriented segments of gluteus minimus, and three segments of gluteus medius. The guidelines were developed with reference to anatomical reports, cadaveric observation and real-time ultrasound imaging in vivo. Five cadaveric gluteal regions were marked for intramuscular electrode insertions based on these guidelines. Intramuscular electrodes were inserted into the marked regions of gluteus minimus (2×) and gluteus medius (3×) with the aid of a 15 cm biopsy needle. Systematic dissection revealed that electrodes were successfully inserted into uniquely oriented segments of gluteus minimus and medius. The orientation of fascicles surrounding each electrode was also consistent with segmental descriptions in past anatomical research. The findings of this research suggest that the guidelines described may be used to assess the functional role of segments within gluteus minimus and medius in health and dysfunction using EMG. Finally, electromyographers intent on investigating the role of posterior gluteus minimus must be cautious of the superior gluteal neurovascular bundle.


Catalytic processes monitored at the nanoscale with tip-enhanced Raman spectroscopy.
Heterogeneous catalysts play a pivotal role in the chemical industry, but acquiring molecular insights into functioning catalysts remains a significant challenge. Recent advances in micro-spectroscopic approaches have allowed spatiotemporal information to be obtained on the dynamics of single active sites and the diffusion of single molecules. However, these methods lack nanometre-scale spatial resolution and/or require the use of fluorescent labels. Here, we show that time-resolved tip-enhanced Raman spectroscopy can monitor photocatalytic reactions at the nanoscale. We use a silver-coated atomic force microscope tip to both enhance the Raman signal and to act as the catalyst. The tip is placed in contact with a self-assembled monolayer of p-nitrothiophenol molecules adsorbed on gold nanoplates. A photocatalytic reduction process is induced at the apex of the tip with green laser light, while red laser light is used to monitor the transformation process during the reaction. This dual-wavelength approach can also be used to observe other molecular effects such as monolayer diffusion.


Quality and equity of care in U.S. hospitals.
Nearly every U.S. hospital publicly reports its performance on quality measures for patients who are hospitalized for acute myocardial infarction, heart failure, or pneumonia. Because performance rates are not reported according to race or ethnic group, it is unclear whether improvements in equity of care have accompanied aggregate improvements in health care quality over time. We assessed performance rates for quality measures covering three conditions (six measures for acute myocardial infarction, four for heart failure, and seven for pneumonia). These rates, adjusted for patient- and hospital-level covariates, were compared among non-Hispanic white, non-Hispanic black, and Hispanic patients who received care between 2005 and 2010 in acute care hospitals throughout the United States. Adjusted performance rates for the 17 quality measures improved by 3.4 to 57.6 percentage points between 2005 and 2010 for white, black, and Hispanic adults (P<0.001 for all comparisons). In 2005, as compared with adjusted performance rates for white patients, adjusted performance rates were more than 5 percentage points lower for black patients on 3 measures (range of differences, 12.3 to 14.2) and for Hispanic patients on 6 measures (5.6 to 14.5). Gaps decreased significantly on all 9 of these measures between 2005 and 2010, with adjusted changes for differences between white patients and black patients ranging from -8.5 to -11.8 percentage points and from -6.2 to -15.1 percentage points for differences between white patients and Hispanic patients. Decreasing differences according to race or ethnic group were attributable to more equitable care for white patients and minority patients treated in the same hospital, as well as to greater performance improvements among hospitals that disproportionately serve minority patients. Improved performance on quality measures for white, black, and Hispanic adults hospitalized for acute myocardial infarction, heart failure, or pneumonia was accompanied by increased racial and ethnic equity in performance rates both within and among U.S. hospitals. (Funded by the Centers for Medicare and Medicaid Services and the Veterans Affairs Health Services Research and Development Career Development Program.).


Importance of Translocon Subunit Tic56 for rRNA Processing and Chloroplast Ribosome Assembly.
Toc159-containing complexes at the outer chloroplast envelope membrane form stable supercomplexes with a 1-MD translocon at the inner chloroplast envelope membrane of which Tic56 is one essential subunit. While the single mutants tic56-1 and ppi2 (toc159) have an albino phenotype and are able to grow heterotrophically, we find the double mutant to be embryo lethal. Comprehensive quantitative proteome profiling with both single mutants in combination with GeneChip analyses identified a posttranscriptional defect in the accumulation of plastid ribosomal proteins and diminished expression of plastid encoded proteins. In the tic56-1 mutant, the assembly of functional ribosomes is furthermore hampered by a processing defect of the plastid 23S rRNA. Spectinomycin-treatment of wild-type plants phenocopies the molecular phenotype of plastid proteome accumulation in tic56-1 and to a smaller degree also ppi2 plastids, suggesting that a defect in plastid translation is largely responsible for the phenotype of both import mutants. Import experiments with the tic56-3 mutant revealed no significant defect in the import of small ribosomal protein 16 in the absence of full-length Tic56, suggesting that the defect in ribosome assembly in tic56-1 may be independent of a function of Tic56 in protein import. Our data establish a previously unknown link between plastid protein import, the processing of plastid rRNAs, and the assembly of plastid ribosomes and provide further knowledge on the function of the translocon components and the molecular basis for their albino phenotype.


Distribution of Campylobacter jejuni isolates from turkey farms and different stages at slaughter using pulsed-field gel electrophoresis and flaA-short variable region sequencing.
The aim of this study was to assess the diversity of thermotolerant Campylobacter spp. isolated from turkey flocks at six rearing farms 1-2 weeks prior to slaughter (360 faecal swab samples) and from 11 different stages at the slaughterhouse (636 caecal, environmental, neck skin and meat samples). A total of 121 Campylobacter isolates were identified to species level using a multiplex PCR assay and were typed by pulsed-field gel electrophoresis (PFGE) and flaA-short variable region (SVR) sequencing. All Campylobacter isolates were identified as Campylobacter jejuni. PFGE analysis with KpnI restriction enzyme resulted in 11 PFGE types (I-XI) and flaA SVR typing yielded in nine flaA-SVR alleles. The Campylobacter-positive turkey flocks A, C and E were colonized by a limited number of Campylobacter clones at the farm and slaughter. The present study confirms the traceability of flock-specific strains (PFGE types I, V and IX; flaA types 21, 36 and 161) from the farm along the entire processing line to meat cuts. It seems that stress factors such as high temperature of the defeathering water (54-56 °C), drying of the carcass skin during air chilling (24 h at 2 °C), and oxygen in the air could not eliminate Campylobacter completely. Campylobacter-negative flocks became contaminated during processing by the same subtypes of Campylobacter introduced into the slaughter house by preceeding positive flocks even if they were slaughtered on subsequent days. Proper and efficient cleaning and disinfection of slaughter and processing premises are needed to avoid cross-contamination, especially in countries with a low prevalence of Campylobacter spp. The majority of flaA SVR alleles displayed a distinct association with a specific PFGE type. However, a linear relationship for all strains among both typing methods could not be established. To specify genetic relatedness of strains, a combination of different genotyping methods, is needed.


4-Nonylphenol in sewage sludge: accumulation of toxic metabolites from nonionic surfactants.
Anaerobically treated sewage sludge was found to contain extraordinarily high concentrations of 4-nonylphenol, a metabolite derived from nonionic surfactants of the nonylphenol polyethoxylate type. Concentrations in activated sewage sludge, in mixed primary and secondary sludge, and in aerobically stabilized sludge were substantially lower, suggesting that the formation of 4-nonylphenol is favored under mesophilic anaerobic conditions. Because 4-nonylphenol may be highly toxic to aquatic life, further research is needed on the fate of 4-nonylphenol after sludge is disposed of in the environment.


Vitamin D is associated with degree of disability in patients with fully ambulatory relapsing-remitting multiple sclerosis.
Vitamin D deficiency is a recognized risk factor for multiple sclerosis (MS) and is associated with increased disease activity. It has also been proposed that the lower the vitamin D levels are, the higher is the handicap. To refine the links between vitamin D insufficiency and disability in MS patients, a retrospective cohort analysis was performed including 181 patients prospectively followed without previous vitamin D supplementation, and age, gender, age at MS onset, MS type, MS activity, Expanded Disability Status Scale (EDSS) were analysed in correlation with plasma vitamin D levels. Vitamin D levels were significantly higher in relapsing-remitting MS than in progressive forms of MS in multivariate analyses adjusted for age, ethnicity, gender, disease duration and season (P = 0.0487). Overall, there was a negative correlation between vitamin D level and EDSS score (P = 0.0001, r = -0.33). In relapsing-remitting MS, vitamin D levels were only correlated with disability scores for EDSS < 4 (P = 0.0012). Patients with >20 ng/ml of vitamin D were 2.78 times more likely to have an EDSS < 4 (P = 0.0011, 95% confidence interval 1.49-5.00). Data support previous work suggesting that vitamin D deficiency is associated with higher risk of disability in MS. Vitamin D levels also correlated with the degree of disability in fully ambulatory patients with relapsing-remitting MS. These additional results support the pertinence of randomized controlled trials analysing the interest of an early vitamin D supplementation in MS patients to influence evolution of disability.


Effects of irradiation on Planococcus minor (Hemiptera: Pseudococcidae).
Irradiation has been recognized and endorsed as a potential phytosanitary measure that could be an alternative to current quarantine treatments. Dosages of 50, 100, 150, 200, and 250 Gy were used to irradiate three different life stages (eggs, immatures, and adults) of Planococcus minor (Maskell) (Hemiptera: Pseudococcidae), focusing on females due to its parthenogenesis ability, with an aim to find the most tolerant stage and the most optimal dose to control P. minor. Cobalt 60 was the source of irradiation used. Irradiation of 150-250 Gy has a significant effect on all life stages of P. minor, decreasing its survival rate, percentage of adult reproduction, oviposition, and fertility rate. The adult was the most tolerant life stage in both mortality and fertility rate. All the different irradiated target life stage groups oviposited eggs, but none of the F2 eggs hatched at the most optimal dosage of 150-250 Gy.


A push-and-pull model for allosteric anion binding in cage complexes.
A series of electronic structure calculations has been carried out on an artificial anion binding host. The compound with four Pd(II) cations and a total of eight bis-monodentate pyridyl ligands forms by self-assembly an interpenetrated double cage with three binding pockets. Through the use of a simple push-and-pull model connecting the potentials of the different sites, we are able to explain the allosteric effect observed in anion binding. Two factors seem to be particularly significant in the latter, namely the flatness of the potential in each binding pocket as well as the length of the ligand. Our results are found to be in excellent agreement with the experimentally observed structures.


Mechanical properties of the sodium montmorillonite interlayer intercalated with amino acids.
Nanosized montmorillonite clay dispersed in small amounts in polymer results in polymer nanocomposites having superior engineering properties compared to those of the native polymer. These nanoinclusions are created by treating clay with an organic modifier which makes clay organophilic and results in intercalation or exfoliation of the montmorillonite. The modifiers used are usually long carbon chains with alkylammonium or alkylphosphonium cations. In this work, we have investigated the use of some alternative molecules which can act as modifiers for clay composites using clay for reinforcing a matrix of biopeptides or proteins. Such composites have potential applications in the fields of biomedical engineering and pharmaceutical science. In this work, the amino acids arginine and lysine are used as modifiers. The intercalation and mechanical behavior of the interlayer spacing with these amino acids as inclusions under compression and tension are studied using molecular dynamics simulations. Significant differences in the responses are observed. This work also provides an insight into the orientation and interaction of amino acids in the interlayer under different stress paths.


Targeting the underlying pathophysiology in X-linked hypophosphatemic rickets in adults.
X-linked hypophosphatemic rickets (XLHR) is a life-long phosphate waste disorder that presents in early childhood with lower limb deformities, stunted growth, and bone and joint pain. In adults, osteomalacia and fractures may develop, aggravating bone and joint pain, stiffness, and disability. A 50-year-old woman with XLHR was referred to Rheumatology for incapacitating pain in her left lower limb with gait impairment. A pseudofracture was identified in the radiography of long bones, and secondary hyperparathyroidism was also observed. Treatment was optimized, and marked clinical improvement occurred. The authors review and discuss the underlying pathophysiology of this disease and its adequate management.


SARS-CoV-2 Variant Pathogenesis Following Primary Infection and Reinfection in Syrian Hamsters.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of COVID-19, has evolved into multiple variants. Animal models are important to understand variant pathogenesis, particularly for variants with mutations that have significant phenotypic or epidemiological effects. Here, cohorts of naive or previously infected Syrian hamsters (Mesocricetus auratus) were infected with variants to investigate viral pathogenesis and disease protection. Naive hamsters infected with SARS-CoV-2 variants had consistent clinical outcomes, tissue viral titers, and pathology, while hamsters that recovered from initial infection and were reinfected demonstrated less severe clinical disease and lung pathology than their naive counterparts. Males had more frequent clinical signs than females in most variant groups, but few sex variations in tissue viral titers and lung pathology were observed. These findings support the use of Syrian hamsters as a SARS-CoV-2 model and highlight the importance of considering sex differences when using this species. IMPORTANCE With the continued circulation and emergence of new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants, understanding differences in the effects between the initial infection and a subsequent reinfection on disease pathogenesis is critical and highly relevant. This study characterizes Syrian hamsters as an animal model to study reinfection with SARS-CoV-2. Previous infection reduced the disease severity of reinfection with different SARS-CoV-2 variants.


The 5-HT(1A) receptor knockout mouse and anxiety.
The 5-HT(1A) receptor has been implicated in the modulation of anxiety processes, mainly via pharmacological experiments. The recent production, in three independent research groups, of 5-HT(1A) receptor knockout (R KO) mice in three different genetic backgrounds (C57BL/6J, 129/Sv, Swiss-Webster) led to the intriguing finding that all mice, independent from the genetic background strain from which the null mutants were made, showed an "anxious" phenotype compared to corresponding wild-type mice. The present paper reviews the behavioral findings in these three KO lines and focuses on new findings in the 129/Sv-KO mice. These mice were more anxious or stress-prone only under specific conditions (high stress) and not as broadly as suggested from the initial studies. The 5-HT(1A) R KO made in the Swiss-Webster background displays disturbances in the GABA(A)-benzodiazepine (BZ) receptor system in the brain, including downregulation of GABA(A) alpha1 and alpha2 subunits in the amygdala. In contrast, the GABA(A)-BZ receptor system seems to function normally in the 5-HT(1A) R KO in the 129/Sv background suggesting that changes in the GABA(A)-BZ receptor system may not be a prerequisite for anxiety but rather could have a modifying effect on this phenotype. It can be concluded that the constitutive absence of the 5-HT(1A) receptor gene and receptor leads to a more "anxious" mouse, dependent on the stress level but independent from the strain. Depending on the genetic background, this null mutation may be associated with changes in GABA(A)-ergic neurotransmission. It is as yet unclear which mechanisms are involved in this intriguing differentiation.


Computational Approach to Molecular Catalysis by 3d Transition Metals: Challenges and Opportunities.
Computational chemistry provides a versatile toolbox for studying mechanistic details of catalytic reactions and holds promise to deliver practical strategies to enable the rational in silico catalyst design. The versatile reactivity and nontrivial electronic structure effects, common for systems based on 3d transition metals, introduce additional complexity that may represent a particular challenge to the standard computational strategies. In this review, we discuss the challenges and capabilities of modern electronic structure methods for studying the reaction mechanisms promoted by 3d transition metal molecular catalysts. Particular focus will be placed on the ways of addressing the multiconfigurational problem in electronic structure calculations and the role of expert bias in the practical utilization of the available methods. The development of density functionals designed to address transition metals is also discussed. Special emphasis is placed on the methods that account for solvation effects and the multicomponent nature of practical catalytic systems. This is followed by an overview of recent computational studies addressing the mechanistic complexity of catalytic processes by molecular catalysts based on 3d metals. Cases that involve noninnocent ligands, multicomponent reaction systems, metal-ligand and metal-metal cooperativity, as well as modeling complex catalytic systems such as metal-organic frameworks are presented. Conventionally, computational studies on catalytic mechanisms are heavily dependent on the chemical intuition and expert input of the researcher. Recent developments in advanced automated methods for reaction path analysis hold promise for eliminating such human-bias from computational catalysis studies. A brief overview of these approaches is presented in the final section of the review. The paper is closed with general concluding remarks.


Intraoperative near infrared fluorescence imaging for the assessment of the gastric conduit.
Esophagectomy remains a mainstay of multi-modality therapy in the treatment of malignant disease, as well as selected benign conditions. Anastomotic and conduit complications remain the Achilles' heel of these operations, contributing to major morbidity and mortality. Adequate vascular perfusion of the gastric conduit is vital to avoid these complications, with surgeon observational assessment the mainstay of determining the vascular health of the gastric conduit. Rates of morbidity remain significant, and technologies aimed at better assessing relative tissue perfusion and ischemia are increasingly under investigation and utilized. One such technique is the use of intraoperative near-infrared fluorescence imaging to directly assess these parameters. The application of this technique has shown promise in perfusion assessment during esophagectomy, and potential reduction in anastomotic complications.


Conization and healthcare use: a population-based register study.
The aim of this study was to assess whether negative psychological consequences of conization reported in questionnaire studies translated into increased use of the healthcare services that could relieve such symptoms. This was a population-based register study comparing women undergoing conization with a control group of women with normal cytology results. Data were derived from Danish registers. Using the difference-in-differences method, we measured contacts with general practitioners (GPs), hospitals, psychiatrist/psychologists, and use of anxiolytic and antidepressant prescription drugs over 5 years 'before' and 'after' the conization in the study group, and in comparable periods in the control group. During the 'before' period, women who later had a conization had greater contact with GPs and hospitals, and slightly more contact with psychiatrist/psychologists, than control women. In both groups, healthcare use increased significantly from the 'before' to the 'after' period. For contacts with GPs and hospitals, the increase was significantly larger for the conization group than for the control group, but this could be attributed to the standard postconization follow-up process. In the 'before' period, women who later had a conization used fewer drugs than women of the control-group, but their drug use increased similarly over time. The conization event did not result in an increased use of the healthcare services that could relieve potential negative side effects. However, women who underwent a conization seemed to constitute a select group as they already used GPs and hospitals more frequently, and anxiolytic and antidepressant drugs less frequently, than other women in the years 'before' the conization event.


Increasing evidence of the role of gene flow in animal evolution: hybrid speciation in the yellow-rumped warbler complex.
In this issue of Molecular Ecology, Brelsford et al. (2011) present strong evidence for a case of hybrid speciation within the yellow-rumped warbler complex. Although homoploid hybrid speciation has now been documented in many animals (Mallet 2007), it seems rare in tetrapods (Mavárez & Linares 2008) and it has barely even been mentioned in birds (Price 2008). Brelsford and colleagues thus present the first detailed molecular evidence suggesting that hybrid speciation can occur in birds. Brelsford et al. (2011) posit that Audubon's warbler (Dendroica auduboni) constitutes a hybrid species originating from the admixture of two distinct parental lineages, represented today by myrtle warbler (D. coronata) and black-fronted warbler (D. nigrifrons). The authors present three major lines of molecular evidence suggesting that this is not simply a case of a hybrid swarm or limited introgression.


Somatostatin potentiation of insulin-induced glucose uptake in normal individuals.
The ability of somatostatin (SRIF) to enhance insulin-stimulated glucose uptake was evaluated during clamp studies in normal individuals and patients with non-insulin-dependent diabetes mellitus (NIDDM). The results demonstrated that glucose uptake at insulin levels of approximately 100 microU/ml was significantly greater (P less than 0.001) in normal individuals in response to insulin plus SRIF as compared with insulin alone. In contrast, SRIF did not enhance insulin-stimulated glucose uptake in patients with NIDDM. Measurements were also made of the relative ability of insulin as compared with insulin plus SRIF to suppress C-peptide and glucagon concentrations during the clamp studies. The results of these experiments showed that SRIF did not potentiate the ability of insulin to suppress C-peptide concentrations in normal subjects but did in patients with NIDDM. However, plasma glucagon levels were reduced to a greater degree when SRIF was added to insulin in both normal and diabetic individuals.


Posterior fast craniopharyngioma.
A case is presented of a craniopharyngioma arising in and wholly confined to the posterior fossa in a 23-year-old man. The origin of this unusually located tumour is discussed and comparison made with previously reported ectopic craniopharyngiomas. Most of the cases appeared to be either direct extensions from the suprasellar region or recurrences in locations distant from the original previously treated suprasellar tumours. We found no evidence in this case to relate the tumour to a sellar/suprasellar origin and the craniopharyngioma is believed to have originated in the posterior fossa.


New trends in asthma.
Asthma, better termed hyperactive airway disease, is characterized by increased responsiveness of the tracheo-bronchial tree to exogenous and endogenous stimuli. The hallmark of this illness is widespread inflammation and narrowing of the tracheo-bronchial tree. This is manifested clinically by paroxysms of dyspnea, wheezing and cough which generally occur together. Treatment involves addressing the bronchospastic as well as the inflammatory components of asthma.


Bovine intermediate pituitary alpha-amidation enzyme: preliminary characterization.
A secretory granule-associated enzymatic activity that converts mono-[125I]-D-Tyr-Val-Gly into mono-[125I]-D-Tyr-Val-NH2 has been studied. The activity is primarily soluble and shows optimal activity at pH 7 to pH 8. Amidation activity was stimulated 9-fold by addition of optimal amounts of copper (3 microM). In the presence of optimal copper, ascorbate stimulated the reaction 7-fold; none of the other reduced or oxidized cofactors tested was as effective. Taking into account the dependence of the reaction on ascorbate and molecular oxygen and the production of glyoxylate [2], it is suggested that the alpha-amidation enzyme is a monooxygenase. Lineweaver Burk plots with D-Tyr-Val-Gly as the varied substrate demonstrated Michelis-Menten type kinetics with the values of Km and Vmax increasing with the addition of ascorbate to the assay. A variety of peptides ending with a COOH-terminal Gly residue act as inhibitors of the reaction. Two synthetic peptides, gamma 2MSH and ACTH(1-14), with carboxyl termini similar to the presumed physiological substrates for the enzyme, act as competitive inhibitors with similar K1 values. It is likely that this secretory granule alpha-amidation activity is involved in the physiological biosynthetic alpha-amidation of a wide range of bioactive peptides.