Omega-3 Polyunsaturated Fatty Acids Mitigate Palmitate-Induced Impairments in Skeletal Muscle Cell Viability and Differentiation.
Accumulation of excess saturated free fatty acids such as palmitate (PAL) in skeletal muscle leads to reductions in mitochondrial integrity, cell viability and differentiation. Omega-3 polyunsaturated fatty acids (n-3 PUFAs) such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) counteract PAL-induced lipid accumulation. EPA and DHA, as well as the n-3 PUFA docosapentaenoic acid (DPA), may therefore mitigate PAL-induced lipotoxicity to promote skeletal muscle cell survival and differentiation. C2C12 myoblasts were treated with 50 μM EPA, DPA, or DHA in the absence or presence of 500 μM PAL for 16 h either prior to myoblast analysis or induction of differentiation. Myoblast viability and markers of apoptosis, endoplasmic reticulum (ER) stress and myotube differentiation capacity were investigated using fluorescence microscopy and immunoblotting. High-resolution respirometry was used to assess mitochondrial function and membrane integrity. PAL induced cell death via apoptosis and increased protein content of ER stress markers BiP and CHOP. EPA, DPA, and DHA co-treatment maintained cell viability, prevented PAL-induced apoptosis and attenuated PAL-induced increases in BiP, whereas only DPA prevented increases in CHOP. PAL subsequently reduced protein content of the differentiation marker myogenin and inhibited myotube formation, and all n-3 PUFAs promoted myotube formation in the presence of PAL. Furthermore, DPA prevented PAL-induced release of cytochrome c and maintained mitochondrial integrity. These findings demonstrate the n-3 PUFAs EPA, DPA and DHA elicit similar protective effects against PAL-induced impairments in muscle cell viability and differentiation. Mechanistically, the protective effects of DPA against PAL lipotoxicity are attributable in part to its ability to maintain mitochondrial respiratory capacity via mitigating PAL-induced loss of mitochondrial membrane integrity.


[Severe primary liver abscess and septic pulmonary embolism due to Klebsiella pneumoniae with hypermucoviscosity phenotype].
A 70-year-old man with diabetes mellitus seen for fever, right chest pain, and right-lung field consolidation on chest X-ray was found in thoracoabdominal computed tomography (CT) to have variable-sized nodules in both lung fields and multiple low-density hepatic areas. On physical examination, his pulse was 145 beats per minute and blood pressure 92/68mmHg, indicating a preshock state. Laboratory tests showed elevated WBC of 15,200/microL, serum-C-reactive protein (CRP) of 34.4 mg/dL, and a decreased platelet count of 16,000/microL. Suspecting liver abscesses complicated by a septic pulmonary embolism, we immediately conducted percutaneous transhepatic abscess drainage (PTAD). Liver abscess blood culture and drainage fluid grew the Klebsiella pneumoniae hypermucoviscosity phenotype, carrying the rmpA gene. Although the man had been in critical condition on admission, broad-spectrum antibiotics and PTAD treatment improved his clinical condition to where he could be discharged without problem.


Immunologic phenomena in the effusive form of feline infectious peritonitis.
The effusive form of feline infectious peritonitis (FIP) was reproduced by injecting 12- to 16-week-old kittens intraperitoneally with a cell-free inoculum derived from the tissues of infected cats. The kittens used for the study were either positive for FIP virus-reacting antibodies before inoculation or they were seronegative. Seropositive kittens were obtained from a cattery where the natural infection was enzootic, and seronegative kittens were obtained from a specific-pathogen-free cattery. Only about half the kittens that were seronegative before inoculation developed disease or serum antibodies to the tissue-derived virus. Seronegative kittens that developed disease showed no signs of illness until 8 to 10 days after inoculation, and they lived for 7 to 14 days after clinical signs appeared. The onset of clinical disease coincided with the appearance of serum antibodies. In contrast, all of the seropositive kittens became ill within 36 to 48 hours after inoculation, and died within 5 to 7 days. If seronegative kittens were treated with immune serum or immunoglobulin (Ig)G, they developed disease with the same frequency, acuteness, and severity as seropositive kittens. Foci of hepatitis and serositis in seropositive kittens contained viral antigen, IgG bound to antigen, and complement. Serum complement activity also decreased several days before death in seropositive kittens inoculated with tissue-derived FIP virus. The temporal relationship of clinical disease and the appearance of serum antibodies, the more acute and severe nature of the disease produced in seropositive kittens, and the presence of antibody and complement in the lesions indicated that effusive FIP is immunologically mediated.


Dual effect in the mechanism of action of anti-influenza virus amino acid analogues.
In comparative research studies to clarify the mechanism governing the effects of anti-influenza amino acid analogues, it was established that amino acid analogues exhibit definite chemotherapeutic properties. They possess dual activity capable of inhibiting multiplication and pulmonary consolidation induced by influenza virus infection.


Pesticides.
There is growing evidence about the influence of chemical exposures on specific molecular systems and mechanisms involved in cognitive and mental function. Evidence is also emerging about the negative impact of these chemical exposures on mental health, including depression, suicide, and other risks. Despite the growing appreciation of these factors, however, little attention has been paid to the ethical and social implications of their interactions. Drawing on recent work that argues for an environmental neuroethics approach that explicitly brings together ethics, environment, and conditions of the central nervous system, this article focuses on these critical issues for pesticides specifically.


Perception of movement and shape in Alzheimer's disease.
Alzheimer's disease is the most prevalent cause of abnormal cognitive decline in older adults and commonly affects visual function. Recent evidence suggests that Alzheimer's disease impairs the processing of visual motion, but these conclusions are based on conflicting results in the few cases studied, and the processing of complex motion images has not been investigated. In the present study of motion processing in Alzheimer's disease we assessed visual functions in 63 adults: 41 with mild to moderate Alzheimer's disease (mean age 72.3 years) and 22 without dementia (mean age 71.7 years). Processing of motion cues was tested with computer animation sequences known as random-dot cinematograms, which resemble the stimuli used to define motion processing deficits in primates with lesions of cortical area MT. Results showed that participants with Alzheimer's disease required significantly greater thresholds for perceiving shapes defined by motion cues compared with participants without dementia (P = 0.0005). There were no significant differences between the two groups (P < 0.05) in static visual acuity, static spatial contrast sensitivity and, surprisingly, dynamic visual acuity, which was normal, and motion direction discrimination, which was relatively spared. We conclude that mild to moderate Alzheimer's disease has significant effects on the perception of structure from motion with relative sparing of motion direction discrimination. We cannot exclude a contribution by retinal pathology, but retinal dysfunction alone cannot explain the pattern of defects we observed. The complex motion image processing deficit we identified is likely to have a cerebral basis and has the potential to affect navigation and the recognition of objects in relative motion, as encountered during walking and automobile driving.


[Genetic structure and the load of hereditary diseases in five populations of Arkhangel'skaia region].
A population and medical genetic investigation was performed in a number of raions in the Arkhangel' skaya oblast. Random inbreeding coefficients were 0.000358 and 0.000361 in the Vinogradovskii and Krasnoborskii raions. Malecot's local inbreeding coefficients were 0.000565 and 0.000472, respectively. The endogamy indices were 0.37 and 0.54, respectively. In the urban population, the loads of autosomal dominant, autosomal recessive, and X-linked pathology were 1.01 and 0.98 per 1000 individuals, and 0.29 per 1000 men; in the rural population, they were 1.22, 1.55, and 1.08, respectively. In the populations studied, the hereditary pathology spectrum is described.


Revisiting life strategy concepts in environmental microbial ecology.
Microorganisms are physiologically diverse, possessing disparate genomic features and mechanisms for adaptation (functional traits), which reflect on their associated life strategies and determine at least to some extent their prevalence and distribution in the environment. Unlike animals and plants, there is an unprecedented diversity and intractable metabolic versatility among bacteria, making classification or grouping these microorganisms based on their functional traits as has been done in animal and plant ecology challenging. Nevertheless, based on representative pure cultures, microbial traits distinguishing different life strategies had been proposed, and had been the focus of previous reviews. In the environment, however, the vast majority of naturally occurring microorganisms have yet to be isolated, restricting the association of life strategies to broad phylogenetic groups and/or physiological characteristics. Here, we reviewed the literature to determine how microbial life strategy concepts (i.e. copio- and oligotrophic strategists, and competitor-stress tolerator-ruderals framework) are applied in complex microbial communities. Because of the scarcity of direct empirical evidence elucidating the associated life strategies in complex communities, we rely heavily on observational studies determining the response of microorganisms to (a)biotic cues (e.g. resource availability) to infer microbial life strategies. Although our focus is on the life strategies of bacteria, parallels were drawn from the fungal community. Our literature search showed inconsistency in the community response of proposed copiotrophic- and oligotrophic-associated microorganisms (phyla level) to changing environmental conditions. This suggests that tracking microorganisms at finer phylogenetic and taxonomic resolution (e.g. family level or lower) may be more effective to capture changes in community response and/or that edaphic factors exert a stronger effect in community response. We discuss the limitations and provide recommendations for future research applying microbial life strategies in environmental studies.


Near perfect focusing through multimode fibres.
Holographic, multimode fibre (MMF) based endoscopes envision high-quality in-vivo imaging inside previously inaccessible structures of living organisms, amongst other perspective applications. Within these instruments, a digital micro-mirror device (DMD) is deployed in order to holographically synthesise light fields which, after traversing the multimode fibre, form foci at desired positions behind the distal fibre facet. When applied in various imaging modalities, the purity and sharpness of the achieved foci are determinant for the imaging performance. Here we present diffraction-limited foci, which contain in excess of 96% of optical power delivered by the fibre which, to the best of our knowledge, represents the highest value reported to date. Further, we quantitatively study the impact of various conditions of the experimental procedure including input polarisation settings, influence of ghost diffraction orders, light modulation regimes, bias of the calibration camera and the influence of noise.


Availability of primary care health personnel. The States speak out.
The adequacy of the supply of health personnel, and primary care personnel in particular, has been assessed at the aggregate national level and the disaggregate or regional/state perspective. While Federal programs have been successful in expanding the Nation's supply of health care practitioners and alleviating aggregate national shortages in some occupations and specialties, problems of geographic distribution remain. In an effort to obtain information on the adequacy of the supply of health care personnel within each state and jurisdiction, the chief executives were asked to assess their most pressing personnel supply concerns. The two occupations most often cited as being in short supply were primary care physicians and registered nurses. The state assessment of shortages of registered nurses is in concert with national assessments. In contrast, the supply of primary care physicians appears to be adequate if not in excess at the national level, implying that aggregate assessments may camouflage significant regional and state shortages. Disaggregate assessments are essential to derive an appropriate picture of national supply adequacy.


Parental income and mental disorders in children and adolescents: prospective register-based study.
Children with low-income parents have a higher risk of mental disorders, although it is unclear whether other parental characteristics or genetic confounding explain these associations and whether it is true for all mental disorders. In this registry-based study of all children in Norway (n = 1 354 393) aged 5-17 years from 2008 to 2016, we examined whether parental income was associated with childhood diagnoses of mental disorders identified through national registries from primary healthcare, hospitalizations and specialist outpatient services. There were substantial differences in mental disorders by parental income, except for eating disorders in girls. In the bottom 1% of parental income, 16.9% [95% confidence interval (CI): 15.6, 18.3] of boys had a mental disorder compared with 4.1% (95% CI: 3.3, 4.8) in the top 1%. Among girls, there were 14.2% (95% CI: 12.9, 15.5) in the lowest, compared with 3.2% (95% CI: 2.5, 3.9) in the highest parental-income percentile. Differences were mainly attributable to attention-deficit hyperactivity disorder in boys and anxiety and depression in girls. There were more mental disorders in children whose parents had mental disorders or low education, or lived in separate households. Still, parental income remained associated with children's mental disorders after accounting for parents' mental disorders and other factors, and associations were also present among adopted children. Mental disorders were 3- to 4-fold more prevalent in children with parents in the lowest compared with the highest income percentiles. Parents' own mental disorders, other socio-demographic factors and genetic confounding did not fully explain these associations.


Hepatic and renal betaine-homocysteine methyltransferase activity in pigs as affected by dietary intakes of sulfur amino acids, choline, and betaine.
In Exp. 1, young pigs were fed a basal diet containing .17% methionine (Met) (.14% digestible Met), and .48% cystine (.38% digestible cystine) for 14 d (34 to 48 d of age). Treatment additions were .25% DL-Met, .34% betaine, .30% choline, or .25% DL-Met and .34% betaine. Methionine, but not betaine or choline supplementation, increased (P < .05) weight gain and feed efficiency. Hepatic betaine-homocysteine methyltransferase (BHMT) activity was increased (P < .05) by betaine and choline supplementation but was not affected by Met deficiency. Renal BHMT activity was increased (P < .05) by Met deficiency and was further increased (P < .05) by betaine supplementation. In Exp. 2, 10-kg pigs were fed the basal diet from Exp. 1 supplemented with enough DL-Met to bring the total basal Met to .24% (.20% digestible Met). Treatment additions consisted of .20% DL-Met or .34% betaine, and diets were fed for 16 d (34 to 50 d of age). Feed efficiency increased (P < .05) in response to Met, but not to betaine, supplementation. Hepatic BHMT activity increased (P < .05) in response to betaine and Met, but no changes in renal BHMT activity occurred. Although statistically significant changes in hepatic and renal BHMT activity occurred in both experiments, the magnitude of the responses was probably not physiologically important. Therefore, in contrast to previous findings with rats and chicks, it does not seem that hepatic and renal BHMT activity in pigs is influenced substantially by Met deficiency, or by surfeit levels of choline or betaine.


The influence of pelvic adjustment on the posture of female university students.
[Purpose] The purpose of this study was to examine the effects of pelvic adjustment via the Gonstead Technique on the posture of female university students. [Subjects] The subjects of this study were 30 female university students who were equally and randomly allocated to a pelvic adjustment group, an experimental group, and a stretching group, a control group. [Methods] Using BackMapper, their trunk inclination (TIN), trunk imbalance (TIM), pelvic position (PPO), pelvic torsion (PTO), pelvic rotation (PRO), and position of their scapulae (PSA) were evaluated. [Results] The adjustment group obtained significant results in TIM, PPO, PTO, and PSA, while the control group obtained no significant results for any items. [Conclusion] Pelvic adjustment via the Gonstead Technique may be applied as a method to correct the posture of average adults.


Magnetic resonance imaging of vein wall thickness in patients with Behçet's syndrome.
Vascular disease is a serious complication of Behçet's syndrome (BS), occurring in up to 20% of subjects. Superficial thrombophlebitis, deep vein thrombosis, and arterial aneurysm formation are the most common manifestations. Venous thrombosis is thought to result from vessel wall inflammation. This work investigated the potential usefulness of high resolution magnetic resonance imaging (MRI) to identify inflammation in the venous walls in BS subjects. Seven healthy control (HC) subjects and five BS subjects were scanned with 3T MRI (Siemens Skyra). A standard MRI sequence was adapted for use in the venous system. Metronome guided breathing generated a regular respiratory variation of venous blood velocity. The vein wall imaging was triggered at an appropriate delay after the metronome. The popliteal vein was imaged. Vein wall images were ranked based on wall thickness and signal enhancement by two blinded, experienced observers. Popliteal vein rank scores were found to be significantly increased in BS versus HC subjects by the first observer (p(Observer 1)=0.025, p(Observer2)=0.07) and also averaging both observers (p=0.05). The repeated images of each subject gave a degree of variability in results, potentially from drifting response to metronome guidance over the 10 minute scan. MR imaging can detect increased vein wall thickness in BS subjects compared to healthy controls. Variable response to the metronome-guided breathing requires further development.


HPV DeepSeq: An Ultra-Fast Method of NGS Data Analysis and Visualization Using Automated Workflows and a Customized Papillomavirus Database in CLC Genomics Workbench.
Next-generation sequencing (NGS) has actualized the human papillomavirus (HPV) virome profiling for in-depth investigation of viral evolution and pathogenesis. However, viral computational analysis remains a bottleneck due to semantic discrepancies between computational tools and curated reference genomes. To address this, we developed and tested automated workflows for HPV taxonomic profiling and visualization using a customized papillomavirus database in the CLC Microbial Genomics Module. HPV genomes from Papilloma Virus Episteme were customized and incorporated into CLC "ready-to-use" workflows for stepwise data processing to include: (1) Taxonomic Analysis, (2) Estimate Alpha/Beta Diversities, and (3) Map Reads to Reference. Low-grade (n = 95) and high-grade (n = 60) Pap smears were tested with ensuing collective runtimes: Taxonomic Analysis (36 min); Alpha/Beta Diversities (5 s); Map Reads (45 min). Tabular output conversion to visualizations entailed 1-2 keystrokes. Biodiversity analysis between low- (LSIL) and high-grade squamous intraepithelial lesions (HSIL) revealed loss of species richness and gain of dominance by HPV-16 in HSIL. Integrating clinically relevant, taxonomized HPV reference genomes within automated workflows proved to be an ultra-fast method of virome profiling. The entire process named "HPV DeepSeq" provides a simple, accurate and practical means of NGS data analysis for a broad range of applications in viral research.


[Leaching of uranium containing phosphorites with heterotrophic microorganisms].
The problem of heterotrophic leaching of metals was tested with uranium-containing phosphorites under laboratory conditions with regard to possible technical applications. As leaching agents we used the acids of citric and lactic acid fermentation, carried out with a strain of Aspergillus niger in sulfite liquor under different conditions and with lactic acid bacteria in wheye. Up to 12% uranium were soluted with citric acid fermentation of Marocco phosphorite containing 153 ppm U in Aspergillus niger cultures within 27 days.


["Golden proportion" and its application to calculate dentition].
Within an evolutionary process, the nature has created the standard of aesthetics - a "gold proportion" on the basis of which, the parts of human body, to be more exact, teeth and denture correspond to each other and to own parts by the size, which is the ideal precondition for ideal appearance. The charming smile serves as the proof, that teeth in denture are located by a principle of "gold proportion". A "gold proportion" is the corner stone of beauty and it can be applied with success in stomatology. Proportion is the certain ratio between parts, and proportional means a proper correlation of parts among themselves. It is reputed, that knowledge about "gold proportion" Pythagor has got from products of the Egyptian and Babylon scientists. And this is true, proportions of cult constructions, bas-relieves, pyramids in Giza, home appliances and ornaments from Tutanhamon tomb testify, that under their creation the Egyptian masters were guided by a principle of "gold proportion". The facade of ancient Greek temple Parthenon is built by a principle of "gold proportion". During archeological digs of this temple the compasses which sculptors and architects of an ancient world used has been found. The "gold proportion" is mentioned in the work which has reached us "Beginning" the author is the scientist of antique epoch Euclid. In 1509, in Venice the book of Luka Pacholi the "Divine proportion" has been published, its illustration is attributed to Leonardo de Vinci. This work has been recognized as a "Hymn of a gold proportion". In 1885 the German researcher professor Zeising published his work - "Aesthetic researches". When Zeising has received numerical values of piece length, he saw that they coincided with figures of some numerical sequence, which was offered by the great Italian mathematician of Middle Ages Fibonacci (or Leonardo Pisano). In his composition the "Abacus Book" Leonardo Fibonacci showed aforesaid sequence of numbers, by means of which he has explained the formula of duplication of rabbits. The specified sequence scientists have seen in many phenomena of the Universe. In sequence of Fibonacci, in some division of its term on previous, it is received irrational number 1,61803398875.., known as Fibonacci. It is used in the reduced, approximated kind as F = 1,618 or phi = 0,618. Fibonacci also gave a proportional number (...3:2, 5:3, 8:5, 13:8...), where numerator represents the sum of two previous numbers in Fibonacci sequence. And in proportional number, similar to numerical sequence, the result of any ratio will be approximately the same - this an irrational number which asymptotically comes to 1,61803398875.... Two numbers of Fibonacci sequence F = 1,618 or phi = 0,618 or phi = 1:F... represent constants of the same sequence. It is actual in stomatology to use proportional way. For example, in orthopedic stomatology taking into consideration of ratio 5:3, promotes improvement of basic fabrics tension, prolongation of teeth functioning and denture, and also planning of correct redistribution of chewing function. In 60th -70th years the American stomatologists carried out many researches to define what kind of part "gold proportion" takes in this area of medicine. Orthodontist Robert Rikets has established that separate teeth correlate with each other by a principle of "gold proportion". In his works Roberts Rickets has specified, that mentioned data is necessary to be considered in aesthetic stomatology during teeth reconstruction. An eminent stomatologist from California, the surgeon, doctor Shtefan Markvart has found out that the height of lateral incisors corresponds to width of lateral incisors by a principle of "gold proportion". This fact has helped the decision of many aesthetic problems. Knowing the width, it is easy to calculate the height and vice versa. When the width is optimum, it cannot be broken with the purpose of closing of an interval. The principle of "gold proportion" in connection with a denture consists in the following: the central incisors are hardly longer than lateral incisors and are almost equal to the length of canines: and lateral incisors are slightly shorter and thinner than the central incisors. The width and the length of each tooth is measured by compasses and calculated for each patient individually. According to a "gold proportion" fore-teeth as a part of person appearance are in the certain ratio to each other. The basic key for the description and construction of four incisors and two canines considers the size (which is accepted as unit) of the width of a small piece of teeth arch or a lateral incisor. On the basis of a lateral incisor width, by the formula of "gold proportion", it is formed the width of the central incisors and canines, and also the length of a fore-part of a dental arch, and vise versa if the length of a fore-part of a dental arch is known, to achieve an ideal aesthetic condition by a principle of "gold proportion", it is defined the width of all fore-teeth. Teeth measurement is taken to define an initial condition, denture calculation, verification of the received results during teeth construction and estimation of the complete denture reconstruction.


Pax6 defines the di-mesencephalic boundary by repressing En1 and Pax2.
Transcriptional factors and signaling molecules are responsible for regionalization of the central nervous system. In the early stage of neural development, Pax6 is expressed in the prosencephalon, while En1 and Pax2 are expressed in the mesencephalon. Here, we misexpressed Pax6 in the mesencephalon to elucidate the mechanism of the di-mesencephalic boundary formation. Histological analysis, expression patterns of diencephalic marker genes, and fiber trajectory of the posterior commissure indicated that Pax6 misexpression caused a caudal shift of the di-mesencephalic boundary. Pax6 repressed En1, Pax2 and other tectum (mesencephalon)-related genes such as En2, Pax5, Pax7, but induced Tcf4, a diencephalon marker gene. To know how Pax6 represses En1 and Pax2, we ectopically expressed a dominant-active or negative form of Pax6. The dominant-active form of Pax6 showed a similar but more severe phenotype than Pax6, while the dominant-negative form showed an opposite phenotype, suggesting that Pax6 acts as a transcriptional activator. Thus Pax6 may repress tectum-related genes by activating an intervening repressor. The results of misexpression experiments, together with normal expression patterns of Pax6, En1 and Pax2, suggest that repressive interaction between Pax6 and En1/Pax2 defines the di-mesencephalic boundary.


A Brucella melitensis high-temperature-requirement A (htrA) deletion mutant is attenuated in goats and protects against abortion.
It has been previously demonstrated that a Brucella melitensis high-temperature-requirement A (htrA) deletion mutant is more susceptible to oxidative killing in vitro than the parental strain and is attenuated in mice. To evaluate the contribution of the B melitensis HtrA protease to virulence in ruminants, the capacity of the B melitensis htrA mutant RWP5 to produce abortion in goats was compared to that of the virulent parental strain 16M. Experimental infection with strain 16M caused abortion in eight of 12 pregnant nannies, while none of the 12 nannies inoculated with RWP5 aborted. Furthermore, intramuscular injection of fetuses in utero with RWP5 led to colonisation of the fetus with subsequent colonisation of the nanny, but no abortion was observed. Nannies vaccinated with RWP5 showed complete protection against abortion when challenged with 16M during the third trimester of pregnancy. However, these animals were not protected from colonisation by 16M. The results presented here clearly indicate that the B melitensis htrA gene product contributes to pathogenesis in goats, but the utility of B melitensis htrA mutants as vaccines in this host appears to be limited.


Falling hospital and postdischarge mortality following CABG in New South Wales from 2000 to 2013.
To describe changes in mortality among patients undergoing coronary artery bypass grafting (CABG) in New South Wales (NSW) Australia from 2000 to 2013. Patients undergoing CABG were identified from the NSW Admission Patient Data Collection (APDC) registry, linked to the NSW state-wide death registry database. Changes in all-cause mortality over time were observed following stratification of the study cohort into two year groups. We identified 54 767 patients undergoing CABG during the study period. The risk profile of patients increased over time with significant increases in age, comorbidities and concomitant valve surgery (all p < 0.0001). During a median follow-up period of 6 years, a total 12 161 (22.2%) of patients had died. Survival curves and adjusted analyses showed a steady fall in mortality rate: those operated on during 2012-2013 had 40 % lower mortality than those operated on during 2000-2001 (HR 0.61; 95% CI 0.53 to 0.69). This was contributed to both by a fall in mortality both in hospital (HR 0.48, 95% CI 0.37 to 0.62) and postdischarge (HR 0.73; 95% CI 0.61 to 0.86). We report a consistent reduction in medium-term mortality among a large unselected cohort of NSW patients undergoing CABG between 2000 and 2013. This fall is attributable both to an improvement in outcomes in hospital and in the postdischarge period.


Clinical research in aesthetic surgery.
Clinical research in aesthetic surgery cannot use traditional objective measures of surgical success. Present research designs and methods used in aesthetic surgery limit the ability to conduct meaningful clinical research. Outcomes research may be ideally suited for assessing patients in aesthetic surgery. A critical aspect of an outcomes-based approach is to select appropriate instruments for investigations. Widely accepted, standardized methods for assessing outcomes would allow for comparison of surgical techniques and provide a common basis for clinical investigations.


Mapping class I gene sequences in the major histocompatibility complex.
The class I glycoproteins of the major histocompatibility complex (MHC) are products of closely linked genes on the 17th chromosome of the mouse. Four highly homologous cell surface proteins, K, D, L, and Qa-2 (refs 1-4), belonging to this multigene family have been analysed. Several additional members, M, R, TL, and indicated by biochemical data and recent serological data suggest the family is larger. Recent reports using recombinant DNA technology suggest that the class I system is an extensive multigene family. In this report we describe results of experiments using restriction enzyme digestion of DNA from a group of standard, congeneic and MHC recombinant mouse strains followed by hybridization to H-2 cDNA probes. Particular bands of hybridization can be assigned to specific H-2 haplotypes and, therefore, to the MHC regions of the 17th chromosome. A number of the MHC polymorphic bands have been assigned to genetic map positions in the K and D-L regions of the H-2 complex. Additional class I gene sequences map outside the boundaries of the traditional H-2 complex (K, I, S, D), into the Qa-T1a region. At least one sequence is centromeric to the K locus. Further, amino terminal and carboxy terminal probes show different restriction patterns, suggesting that the exons corresponding to the different domains of the class I glycoproteins may have different evolutionary histories.


Mechanisms of physical activity limitation in chronic lung diseases.
In chronic lung diseases physical activity limitation is multifactorial involving respiratory, hemodynamic, and peripheral muscle abnormalities. The mechanisms of limitation discussed in this paper relate to (i) the imbalance between ventilatory capacity and demand, (ii) the imbalance between energy demand and supply to working respiratory and peripheral muscles, and (iii) the factors that induce peripheral muscle dysfunction. In practice, intolerable exertional symptoms (i.e., dyspnea) and/or leg discomfort are the main symptoms that limit physical performance in patients with chronic lung diseases. Furthermore, the reduced capacity for physical work and the adoption of a sedentary lifestyle, in an attempt to avoid breathlessness upon physical exertion, cause profound muscle deconditioning which in turn leads to disability and loss of functional independence. Accordingly, physical inactivity is an important component of worsening the patients' quality of life and contributes importantly to poor prognosis. Identifying the factors which prevent a patient with lung disease to easily carry out activities of daily living provides a unique as well as important perspective for the choice of the appropriate therapeutic strategy.


Multiplex Immunohistochemistry Staining for Paraffin-embedded Lung Cancer Tissue.
Lung cancer is the leading cause of malignant tumor-related morbidity and mortality all over the world, and the complex tumor microenvironment has been considered the leading cause of death in lung cancer patients. The complexity of the tumor microenvironment requires effective methods to understand cell-to-cell relationships in tumor tissues. The multiplex immunohistochemistry (mIHC) technique has become a key tool for inferring the relationship between the expression of proteins upstream and downstream of signaling pathways in tumor tissues and developing clinical diagnoses and treatment plans. mIHC is a multi-label immunofluorescence staining method based on Tyramine Signal Amplification (TSA) technology, which can simultaneously detect multiple target molecules on the same tissue section sample to achieve different protein co-expression and co-localization analysis. In this experimental protocol, paraffin-embedded tissue sections of lung squamous carcinoma of clinical origin were subjected to multiplex immunohistochemical staining. By optimizing the experimental protocol, multiplex immunohistochemical staining of labeled target cells and proteins was achieved, solving the problem of autofluorescence and channel crosstalk in lung tissues. In addition, multiplex immunohistochemical staining is widely used in the experimental validation of tumor-related, high-throughput sequencing, including single-cell sequencing, proteomics, and tissue space sequencing, providing intuitive and visual pathology validation results.


Expression of tumor necrosis factor-alpha and interferon-gamma mRNA in blood cells correlates with depression scores during an acute attack in patients with multiple sclerosis.
Depression is a common problem in multiple sclerosis (MS) and affects about 50% of MS patients. Since a dysregulation of cytokine levels has been implicated in the pathogenesis of MS and alterations in cytokine serum levels have been found in depressive illness, we examined the relationship between depressive symptoms, cytokine mRNA expression levels of Th1-type and Th2-type cytokines and neurological disability among early diagnosed MS patients in a prospective study. Sixteen patients with clinically or laboratory supported MS were assessed using the Beck Depression Inventory (BDI) and the Kurtzke Expanded Disability Status Scale (EDSS). Cytokine mRNA in whole blood was serially determined by a new quantitative polymerase chain reaction (PCR) method. BDI sum scores (2,9 fold) and the expression levels of tumor necrosis factor-alpha (TNF-alpha; 4 fold), interferon-gamma (IFN-gamma; 4,6 fold) and interleukin-10 (IL-10; 6,1 fold) mRNA were increased in MS patients during an acute attack compared to age and sex matched healthy controls. We detected a significant positive correlation between TNF-alpha (r=0.55) and interferon-gamma (r=0.54) mRNA expression and the BDI sum scores during an acute attack in MS patients. At follow-up after 3-6 months, only TNF-alpha mRNA expression was correlated with BDI sum scores (r=0.62 resp. r=0.31). No correlation of the BDI sum scores with Th2-type cytokine mRNA expression for interleukin-4 (IL-4) and interleukin-10 (IL-10) or with the extent of neurological disability was observed. The possible contribution of Th1-type cytokines to the development of depression in MS is discussed.


Guideline for the use of high-level disinfectants and sterilants for reprocessing of flexible gastrointestinal endoscopes.
Glutaraldehyde, 0.2% peracetic acid, 7.5% hydrogen peroxide, 0.08% peracetic acid/1% hydrogen peroxide, and 0.55% ortho-phthalaldehyde are cleared by the FDA for reprocessing flexible gastrointestinal endoscopes. Each product has advantages and disadvantages. All require adherence to published reprocessing protocols in order to maintain the integrity of equipment while providing the public with endoscopic instruments that are safe and effective. All chemicals must be handled with respect. Selection of a product must be weighed against the needs of a particular setting, taking into consideration factors such as compatibility, toxicity, environmental controls and cost.


Flower meristem maintenance by TILLERS ABSENT 1 is essential for ovule development in rice.
Plant development depends on the activity of pluripotent stem cells in meristems, such as the shoot apical meristem and the flower meristem. In Arabidopsis thaliana, WUSCHEL (WUS) is essential for stem cell homeostasis in meristems and integument differentiation in ovule development. In rice (Oryza sativa), the WUS ortholog TILLERS ABSENT 1 (TAB1) promotes stem cell fate in axillary meristem development, but its function is unrelated to shoot apical meristem maintenance in vegetative development. In this study, we examined the role of TAB1 in flower development. The ovule, which originates directly from the flower meristem, failed to differentiate in tab1 mutants, suggesting that TAB1 is required for ovule formation. Expression of a stem cell marker was completely absent in the flower meristem at the ovule initiation stage, indicating that TAB1 is essential for stem cell maintenance in the 'final' flower meristem. The ovule defect in tab1 was partially rescued by floral organ number 2 mutation, which causes overproliferation of stem cells. Collectively, it is likely that TAB1 promotes ovule formation by maintaining stem cells at a later stage of flower development.


RasGAP Promotes Autophagy and Thereby Suppresses Platelet-Derived Growth Factor Receptor-Mediated Signaling Events, Cellular Responses, and Pathology.
Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) make profound contributions to both physiology and pathology. While it is widely believed that direct (PDGF-mediated) activation is the primary mode of activating PDGFRs, the discovery that they can also be activated indirectly begs the question of the relevance of the indirect mode of activating PDGFRs. In the context of a blinding eye disease, indirect activation of PDGFRα results in persistent signaling, which suppresses the level of p53 and thereby promotes viability of cells that drive pathogenesis. Under the same conditions, PDGFRβ fails to undergo indirect activation. In this paper, we report that RasGAP (GTPase-activating protein of Ras) prevented indirect activation of PDGFRβ. RasGAP, which associates with PDGFRβ but not PDGFRα, reduced the level of mitochondrion-derived reactive oxygen species, which are required for enduring activation of PDGFRs. Furthermore, preventing PDGFRβ from associating with RasGAP allowed it to signal enduringly and drive pathogenesis of a blinding eye disease. These results indicate a previously unappreciated role of RasGAP in antagonizing indirect activation of PDGFRβ, define the underlying mechanism, and raise the possibility that PDGFRβ-mediated diseases involve indirect activation of PDGFRβ.


Functional magnetic resonance imaging during urodynamic testing identifies brain structures initiating micturition.
Normal voiding in neurologically intact patients is triggered by the release of tonic inhibition from suprapontine centers, allowing the pontine micturition center to trigger the voiding reflex. Supraspinal mechanisms of voluntary voiding in humans are just beginning to be described via functional neuroimaging. We further elucidated brain activity processes during voiding using functional magnetic resonance imaging in normal females to gain better understanding of normal voiding as well as changes that may occur in voiding dysfunction. We screened 13 healthy premenopausal female volunteers using baseline clinic urodynamics to document normal voiding parameters. We then recorded brain activity via functional magnetic resonance imaging and simultaneous urodynamics, including the pressure flow voiding phase. After motion correction of functional magnetic resonance images we performed activation and connectivity analyses in 10 subjects. Group analysis revealed consistent activation areas, including regions for motor control (cerebellum, thalamus, caudate, lentiform nucleus, red nucleus, supplementary motor area and post-central gyrus), emotion (anterior/posterior cingulate gyrus and insula), executive function (left superior frontal gyrus) and a focal region in the pons. Connectivity analysis demonstrated strong interconnectivity of the pontine micturition center with many short-range and long-range cortical clusters. Our study is one of the first reports of brain activation centers associated with micturition initiation in normal healthy females. Results show activation of a brain network consisting of regions for motor control, executive function and emotion processing. Further studies are planned to create and validate a model of brain activity during normal voiding in women.


Childhood Obesity-Related Mechanisms: MicroRNome and Transcriptome Changes in a Nested Case-Control Study.
Childhood obesity could contribute to adulthood obesity, leading to adverse health outcomes in adults. However, the mechanisms for how obesity is developed are still unclear. To determine the epigenome-wide and genome-wide expression changes related with childhood obesity, we compared microRNome and transcriptome levels as well as leptin protein levels in whole bloods of 12 obese and 24 normal children aged 6 years. miR-328-3p, miR-1301-3p, miR-4685-3p, and miR-6803-3p were negatively associated with all obesity indicators. The four miRNAs were also associated with 3948 mRNAs, and separate 475 mRNAs (185 among 3948 mRNAs) were associated with all obesity indicators. The 2533 mRNAs (64.2%) among the 3948 mRNAs and 286 mRNAs (60.2%) among the 475 mRNAs were confirmed as targets of the four miRNAs in public databases through miRWalk 2.0. Leptin protein was associated with miR-6803-3p negatively and all obesity indicators positively. Using DAVID bioinformatics resources 6.8, top three pathways for obesity-related gene set were metabolic pathways, pathways in cancer, and PI3K-Akt signaling pathway. The top three obesity-related disease classes were metabolic, cardiovascular, and chemdependency. Our results support that childhood obesity could be developed through miRNAs-related epigenetic mechanism and, further, these obesity-related epigenetic changes could control the pathways related with the development of various diseases.


Chronic intermittent high altitude exposure, occupation, and body mass index in workers of mining industry.
The obesity and overweight rates in population exposed to chronic intermittent exposure to high altitudes are not well studied. The aim of the retrospective study was to evaluate whether there are differences in body mass index in different occupation groups working in intermittent shifts at mining industry at high altitude: 3800-4500 meters above sea level. Our study demonstrated that obesity and overweight are common in workers of high altitude mining industry exposed to chronic intermittent hypoxia. The obesity rate was lowest among miners as compared to blue- and white-collar employees (9.5% vs. 15.6% and 14.7%, p=0.013). Obesity and overweight were associated with older age, higher rates of increased blood pressure (8.79% and 5.72% vs. 1.92%), cholesterol (45.8% and 45.6% vs. 32.8%) and glucose (4.3% and 1.26% vs. 0.57%) levels as compared to normal body mass index category (p<0.0001 for all). There were differences in patterns of cholesterol and glucose levels in men and women employees according to occupation type. In conclusion, obesity and overweight rates are prevalent and associated with increase in blood pressure, cholesterol, and glucose levels in workers of mining industry exposed to intermittent high-altitude hypoxia. Therefore, assessment and monitoring of body mass index seems to be essential in those who live and work at high altitudes to supply the correct nutrition, modify risk factors, and prevent related disorders.


Chlamydia pneumoniae infection in adults with chronic cough compared with healthy blood donors.
In a small uncontrolled study, persistent cough has recently been found to be associated with serological evidence of acute Chlamydia pneumoniae infection. In order to assess whether C. pneumoniae plays a role in chronic cough, the prevalence of C. pneumoniae infection in 201 adult patients with chronic cough was compared with the prevalence in 106 healthy blood donors without respiratory tract symptoms in the preceding 3 months. A microimmunofluorescence antibody test was used to determine C. pneumoniae antibodies in the immunoglobulin (Ig)M, IgG and IgA fractions. Further, nasopharyngeal aspirates from the 201 patients were examined for C. pneumoniae deoxyribonucleic acid by polymerase chain reaction (PCR). As judged by serology, nine patients (4%) and one control (1%) had acute C. pneumoniae infection, and 92 patients (46%) and 42 controls (40%) had previous or chronic C. pneumoniae infection. Of the nine patients with acute infection, three were C. pneumoniae PCR positive, and they all had an IgM antibody titre response. The remaining six patients had either an IgG antibody titre of > or =512 (five patients) or an IgA antibody titre of > or =512 (one patient). None of these six patients had detectable IgM antibodies. The mean cough period for the five IgG positive patients (10.8 weeks) was significantly longer than the mean cough period for the remaining patient population (6.4 weeks; p=0.004). It is concluded that Chlamydia pneumoniae infection was not statistically significantly more prevalent in patients with chronic cough than in healthy blood donors, and that Chlamydia pneumoniae appears to have a minor role in patients with chronic cough. Direct detection of Chlamydia pneumoniae by polymerase chain reaction on nasopharyngeal aspirates is highly correlated with detectable immunoglobulin M antibodies, but in the late stages of prolonged cough serological testing of immunoglobulin G and immunoglobulin A may be more beneficial for obtaining a microbiological diagnosis.


The relative responsiveness to inhaled leukotriene E4, methacholine and histamine in normal and asthmatic subjects.
The relative bronchoconstricting potencies of leukotriene E4 (LTE4) methacholine and histamine have been compared in asthmatic and normal subjects. LTE4 responsiveness in asthmatic subjects, as measured by the dose which produced a 35% fall in specific airways conductance (PD35), ranged from 0.06-24.4 nmol (geom mean 4.1 nmol, n = 20). This was significantly less than the PD35 in normal subjects (range 39.0-370 nmol, geom mean 105 nmol, n= 6; p less than 0.001). There was a correlation between LTE4 and methacholine responsiveness (r = 0.84, p less than 0.001) and between LTE4 and histamine responsiveness (r = 0.79, p less than 0.001). LTE4 was 73 times more potent than methacholine and 112 times more potent than histamine in asthmatic subjects. LTE4 was 20 times more potent than methacholine and 58 times more potent than histamine in normal subjects. LTE4 is a potent bronchoconstrictor agent, and LTE4 responsiveness correlates with both histamine and methacholine responsiveness.


Pediatric acute lymphoblastic leukemia.
The outcome for children with acute lymphoblastic leukemia (ALL) has improved dramatically with current therapy resulting in an event free survival exceeding 75% for most patients. However significant challenges remain including developing better methods to predict which patients can be cured with less toxic treatment and which ones will benefit from augmented therapy. In addition, 25% of patients fail therapy and novel treatments that are focused on undermining specifically the leukemic process are needed urgently. In Section I, Dr. Carroll reviews current approaches to risk classification and proposes a system that incorporates well-established clinical parameters, genetic lesions of the blast as well as early response parameters. He then provides an overview of emerging technologies in genomics and proteomics and how they might lead to more rational, biologically based classification systems. In Section II, Drs. Mary Relling and Stella Davies describe emerging findings that relate to host features that influence outcome, the role of inherited germline variation. They highlight technical breakthroughs in assessing germline differences among patients. Polymorphisms of drug metabolizing genes have been shown to influence toxicity and the best example is the gene thiopurine methyltransferase (TPMT) a key enzyme in the metabolism of 6-mercaptopurine. Polymorphisms are associated with decreased activity that is also associated with increased toxicity. The role of polymorphisms in other genes whose products play an important role in drug metabolism as well as cytokine genes are discussed. In Sections III and IV, Drs. James Downing and Cheryl Willman review their findings using gene expression profiling to classify ALL. Both authors outline challenges in applying this methodology to analysis of clinical samples. Dr. Willman describes her laboratory's examination of infant leukemia and precursor B-ALL where unsupervised approaches have led to the identification of inherent biologic groups not predicted by conventional morphologic, immunophenotypic and cytogenetic variables. Dr. Downing describes his results from a pediatric ALL expression database using over 327 diagnostic samples, with 80% of the dataset consisting of samples from patients treated on a single institutional protocol. Seven distinct leukemia subtypes were identified representing known leukemia subtypes including: BCR-ABL, E2A-PBX1, TEL-AML1, rearrangements in the MLL gene, hyperdiploid karyotype (i.e., > 50 chromosomes), and T-ALL as well as a new leukemia subtype. A subset of genes have been identified whose expression appears to be predictive of outcome but independent verification is needed before this type of analysis can be integrated into treatment assignment. Chemotherapeutic agents kill cancer cells by activating apoptosis, or programmed cell death. In Section V, Dr. John Reed describes major apoptotic pathways and the specific role of key proteins in this response. The expression level of some of these proteins, such as BCL2, BAX, and caspase 3, has been shown to be predictive of ultimate outcome in hematopoietic tumors. New therapeutic approaches that modulate the apoptotic pathway are now available and Dr. Reed highlights those that may be applicable to the treatment of childhood ALL.


Preventing the link between SES and high-risk behaviors: "value-added" education, drug use and delinquency in high-risk, urban schools.
We examined whether schools achieving better than expected educational outcomes for their students influence the risk of drug use and delinquency among urban, racial/ethnic minority youth. Adolescents (n = 2,621), who were primarily African American and Hispanic and enrolled in Chicago public schools (n = 61), completed surveys in 6th (aged 12) and 8th (aged 14) grades. Value-added education was derived from standardized residuals of regression equations predicting school-level academic achievement and attendance from students' sociodemographic profiles and defined as having higher academic achievement and attendance than that expected given the sociodemographic profile of the schools' student composition. Multilevel logistic regression estimated the effects of value-added education on students' drug use and delinquency. After considering initial risk behavior, value-added education was associated with lower incidence of alcohol, cigarette and marijuana use; stealing; and participating in a group-against-group fight. Significant beneficial effects of value-added education remained for cigarette and marijuana use, stealing and participating in a group-against-group fight after adjustment for individual- and school-level covariates. Alcohol use (past month and heavy episodic) showed marginally significant trends in the hypothesized direction after these adjustments. Inner-city schools may break the links between social disadvantage, drug use and delinquency. Identifying the processes related to value-added education in order to improve school environments is warranted given the high costs associated with individual-level interventions.


Similarities in the light chains of anti-gamma-globulins showing cross-idiotypic specificities.
A marked homogeneity of the light chains was observed in an analysis of 17 IgM proteins with anti-gamma-globulin activity. The V region subgroups of the light chains were determined by both sequence and antigenic analysis. The latter procedure permitted large scale screening for comparisons with control proteins. The two methods showed general agreement in the determination of Kappa III proteins; all proteins positive by antigenic analysis were also positive by sequence but exceptions were noted in the opposite direction. The anti-gamma-globulins showed by antigenic analysis a 92% incidence of VK III light chains as compared to an incidence of 27% among 81 control proteins without this activity. A similar selection was observed for an antigen (VK III b) which subdivided the kappa III proteins. The major Wa group of anti-gamma-globulins which had been delineated previously on the basis of cross-idiotypic specificity was primarily responsible for the special light-chain selection. All the proteins of this group contain VK III light chains and all were of the VK III b type. Current evidence indicates that additional light-chain similarities were involved in the cross-idiotypic specificity of the Wa group. It thus appears that for the anti-gamma-globulins various levels of selection of light chains are manifest ranging from a preponderance of kappa type, of kappa III subgroup, of kappa III b sub-subgroup and perhaps of still further subdivisions to account for the cross-idiotypic specificity.


Functional renal insufficiency during long-term therapy with captopril and enalapril in severe chronic heart failure.
Renal function was evaluated in 104 patients with severe chronic heart failure whom we treated with captopril or enalapril. Seventy patients showed no change or an improvement in renal function (group A), and 34 patients developed functional renal insufficiency (group B). Before converting-enzyme inhibition, group B patients received higher doses of furosemide (p less than 0.02) and had lower central venous pressures (p less than 0.05) than group A patients. After 1 to 3 months of converting-enzyme inhibition, an excessive reduction in left ventricular filling pressure (to less than 15 mm Hg) or mean arterial pressure (to less than 60 mm Hg) was noted in 28 of 34 (82%) patients in group B but in only 22 of 70 patients in group A (31%) (p less than 0.001). At the end of the study, drug-induced azotemia resolved after a reduction in the dosage of diuretics, despite unaltered treatment with captopril and enalapril. Hence, the deterioration of renal function after converting-enzyme inhibition in heart failure is not a toxic or immunologic reaction to therapy but results from specific hemodynamic events that can be ameliorated by sodium repletion.


Probing the fate of lowest-energy near-infrared metal-centered electronic excited states: CuCl(4)(2-) and IrBr(6)(2-).
Ultrafast transient absorption spectroscopy is used to investigate the radiationless relaxation dynamics of CuCl4(2-) and IrBr6(2-) complexes directly promoted into their lowest-energy excited metal-centered states upon near-infrared femtosecond excitation at 2000 nm. Both the excited CuCl4(2-) (2)E and IrBr6(2-) (2)Ug'(T2g) states undergo internal conversion to the ground electronic states, yet with significantly different lifetimes (55 fs and 360 ps, respectively) despite the fact that the (2)E and (2)Ug'(T2g) states are separated by the same energy gap (∼5000 cm(-1)) from the respective ground state. This difference likely arises from the predominance of the Jahn-Teller effect in a Cu(2+) ion and the spin-orbit coupling effect in an Ir(4+) ion. The approach documented in this work may be used for elucidating the role of low-energy metal-centered states in relaxation cascades of a number of coordination compounds, allowing for design of efficient light-triggered metal complexes.


Polypeptides of varicella-zoster virus (VZV) and immunological relationship of VZV and herpes simplex virus (HSV).
Varicella-zoster virus (VZV), labelled with [35S]methionine or [14C]glucosamine, was purified by centrifugation through sucrose gradients followed by equilibrium centrifugation in CsCl, and analysed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). At least 32 polypeptides ranging in mol. wt. from approx. 280 X 10(3) to 21.5 X 10(3) (280K to 21.5K) and six glycopeptides ranging in mol. wt. from approx. 115K to 45K (gp1 to gp6) were found in the virion. The immunological relationship of VZV and herpes simplex virus (HSV) was investigated. In neutralization (NT) tests, no cross-neutralization was observed between VZV and HSV-1 or -2. In fluorescent antibody staining, however, a cross-reaction was observed between VZV- and HSV-1-infected human embryonic lung (HEL) cells and heterologous antiserum. When cross-reactions were investigated by immunoprecipitation followed by SDS-PAGE, several cross-reacting polypeptides were discovered. Cross-reacting glycopeptides of 64K (gp3) and 55K (gp5) were isolated from VZV-infected cell lysates by affinity column chromatography to immobilized HSV-1 antibodies.


Somatoform disorders: time for a new approach in DSM-V.
DSM-III introduced somatoform disorders as a speculative diagnostic category for somatic symptoms "not explained by a general medical condition." Although retained and enlarged in DSM-IV, somatoform disorders have been the subject of continuing criticism by both professionals and patients. The extended period of preparation for DSM-V offers an important opportunity to reconsider the category of somatoform disorders. Exploration of the diverse aims of a diagnostic classification indicates that the authors must not only address the conceptual and practical problems associated with this category but also reconcile it with the parallel medical descriptive classification of functional symptoms and syndromes. The existing somatoform disorders categories require modification. The authors favor the radical option of the abolition of the categories. Diagnoses currently within somatoform disorders could be redistributed into other groupings, and the disorders currently defined solely by somatic symptoms could be placed on axis III as "functional somatic symptoms and syndromes." Greater use could be made of "psychological factors affecting medical condition" on axis I. The authors suggest supplementing the diagnosis of functional somatic symptoms with a multiaxial formulation. The authors promote a classification of somatic symptoms in DSM-V that is compatible with that used in general medicine and offers new opportunities both for research into the etiology and treatment of symptoms and for the greater integration of psychiatry into general medical practice.


Specific suppression of D-RNA G-quadruplex-protein interaction with an L-RNA aptamer.
G-quadruplexes (G4s) are nucleic acid structure motifs that are of significance in chemistry and biology. The function of G4s is often governed by their interaction with G4-binding proteins. Few categories of G4-specific tools have been developed to inhibit G4-protein interactions; however, until now there is no aptamer tool being developed to do so. Herein, we present a novel L-RNA aptamer that can generally bind to D-RNA G-quadruplex (rG4) structure, and interfere with rG4-protein interaction. Using hTERC rG4 as the target for in vitro selection, we report the shortest L-aptamer being developed so far, with only 25 nucleotides. Notably, this new aptamer, L-Apt.4-1c, adopts a stem-loop structure with the loop folding into an rG4 motif with two G-quartet, demonstrates preferential binding toward rG4s over non-G4s and DNA G-quadruplexes (dG4s), and suppresses hTERC rG4-nucleolin interactions. We also show that inhibition of rG4-protein interaction using L-RNA aptamer L-Apt.4-1c is comparable to or better than G4-specific ligands such as carboxypyridostatin and QUMA-1 respectively, highlighting that our approach and findings expand the current G4 toolbox, and open a new avenue for diverse applications.


The role of phenotype on ventilation and exercise capacity in patients affected by COPD: a retrospective study.
The idea of phenotype in chronic obstructive pulmonary disease (COPD) has evolved in the last decades, and the importance of peculiar treatment strategies has now been acknowledged. Although dyspnea and exercise limitation are hallmarks of COPD, this aspect has never been fully explored in literature in terms of disease phenotype. The aim of the present study was to explore the relevance of clinical COPD phenotypes on exercise ventilation and maximal capacity. In this observational cohort retrospective study we analyzed the data of 50 COPD patients who underwent cardiopulmonary exercise test, categorized as emphysematous (n=29), and non-emphysematous (n=21) according to a previously validated model. We found a significant difference in terms of VE/VCO2 slope (median values 32.4 vs 28.0, p=0.015) and VE/VCO2 ratio at nadir (median values 37 vs. 33, p=0.004), which resulted higher in emphysematous patients, who also presented lower PETCO2 values (median values 32.6 vs 35.6, p=0.008). In a subgroup of 31 tests which met the maximality criteria, emphysematous patients presented a significantly lower work rate at peak (median value 51 vs 72% predicted, p=0.016), and showed a lower peak oxygen consumption, although at the limit of significance (median values of 63 vs 85 % predicted, p=0.051). This study extends our knowledge about the characterization of the COPD phenotypical expression of disease, showing that patients affected by emphysema are more prone to ventilatory inefficiency during exercise, and that this is likely to be an important cause of their overall reduced exercise capacity.


Common dietary supplements for weight loss.
Over-the-counter dietary supplements to treat obesity appeal to many patients who desire a "magic bullet" for weight loss. Asking overweight patients about their use of weight-loss supplements and understanding the evidence for the efficacy, safety, and quality of these supplements are critical when counseling patients regarding weight loss. A schema for whether physicians should recommend, caution, or discourage use of a particular weight-loss supplement is presented in this article. More than 50 individual dietary supplements and more than 125 commercial combination products are available for weight loss. Currently, no weight-loss supplements meet criteria for recommended use. Although evidence of modest weight loss secondary to ephedra-caffeine ingestion exists, potentially serious adverse effects have led the U.S. Food and Drug Administration to ban the sale of these products. Chromium is a popular weight-loss supplement, but its efficacy and long-term safety are uncertain. Guar gum and chitosan appear to be ineffective; therefore, use of these products should be discouraged. Because of insufficient or conflicting evidence regarding the efficacy of conjugated linoleic acid, ginseng, glucomannan, green tea, hydroxycitric acid, L-carnitine, psyllium, pyruvate, and St. John's wort in weight loss, physicians should caution patients about the use of these supplements and closely monitor those who choose to use these products.


Trends in referrals from Croatian family practice in relation to specialists' consultations: a longitudinal observational study from 1995 to 2012.
Drug prescribing and referrals to specialists' consultations, as the indicators of the quality of health care, were not so often monitored in Croatian family medicine. The aims of the study are to determine the trends of referrals from family doctors to specialists' consultations and the trends of specialists' consultations utilization. The study is based on routinely collected data from the Croatian Health Service Yearbooks, 1995-2012. The results indicate that from 1995 to 2012 the visits to family doctors almost doubled, while the number of referrals increased by 60%. The referral rate only slightly increased, from 24 to 29% of the total number of visits. The number of specialists' consultations showed continuous growth and exceeded the number of referrals. The observed trends are not sufficient to explain referral patterns and utilization of specialists' services; therefore, a far more complex research is required.


Morally disengaged and unempathic: do cyberbullies fit these definitions? An exploratory study.
In recent years, the phenomenon of cyberbullying has been gaining scholars' growing interest under various aspects, including its overlap with face-to-face bullying. Nevertheless, its relationships with cognitive and affective empathy, proactive and reactive aggression, and moral disengagement, constructs that proved to be crucial in distinguishing aggressive subjects from their targets and nonaggressive peers in traditional bullying, still represent, to some extent, an unexplored domain. The main purpose of the present exploratory study was to investigate the associations between cyberbullying and the mentioned constructs among Italian adolescents. 819 high-school students (mean age 16.08) were administered a battery of standardized tools, along with Cyberties, a new instrument created to assess the prevalence of (and the type of involvement in) different forms of electronic assaults. Analyses of variance were conducted to compare four roles ("pure" bullies, "pure" victims, bully victims, and noninvolved subjects). Participants who identified themselves as cyberbullies or cyberbully victims showed significantly higher levels of overall moral disengagement and of both types of aggression. Cyberbullies also displayed a lack of affective empathy. Our findings are in line with the ones in extant literature about correlates of traditional and electronic forms of bullying. Implications for prevention strategies are discussed.


Speed dependency in α-motoneuron activity and locomotor modules in human locomotion: indirect evidence for phylogenetically conserved spinal circuits.
Coordinated locomotor muscle activity is generated by the spinal central pattern generators (CPGs). Vertebrate studies have demonstrated the following two characteristics of the speed control mechanisms of the spinal CPGs: (i) rostral segment activation is indispensable for achieving high-speed locomotion; and (ii) specific combinations between spinal interneuronal modules and motoneuron (MN) pools are sequentially activated with increasing speed. Here, to investigate whether similar control mechanisms exist in humans, we examined spinal neural activity during varied-speed locomotion by mapping the distribution of MN activity in the spinal cord and extracting locomotor modules, which generate basic MN activation patterns. The MN activation patterns and the locomotor modules were analysed from multi-muscle electromyographic recordings. The reconstructed MN activity patterns were divided into the following three patterns depending on the speed of locomotion: slow walking, fast walking and running. During these three activation patterns, the proportion of the activity in rostral segments to that in caudal segments increased as locomotion speed increased. Additionally, the different MN activation patterns were generated by distinct combinations of locomotor modules. These results are consistent with the speed control mechanisms observed in vertebrates, suggesting phylogenetically conserved spinal mechanisms of neural control of locomotion.


Simultaneous assay of dihydroxyacetone synthase and transketolase in a methylotrophic yeast grown in continuous culture. A cautionary note.
The methylotrophic yeast Candida boidinii CBS 5777 was grown in continuous culture under carbon limitation on glucose, glucose plus methanol, and methanol as carbon and energy sources. During adaptation from glucose to methanol there was a rapid rise in the specific activities of triokinase, fructose-1,6-bisphosphatase and dihydroxyacetone synthase, which are key enzymes of the xylulose phosphate cycle of formaldehyde fixation. The specific activity of classical transketolase fell during this adaptation. Extracts from carbon-limited C. boidinii contained an enzyme which catalysed oxidation of NADH when some preparations or ribose 5-phosphate were added, which was not a transketolase. This enzyme activity was dependent on an impurity in such ribose 5-phosphate preparations and can be confused with transketolase activity.


Refined solution structure and backbone dynamics of HIV-1 Nef.
The tendency of HIV-1 Nef to form aggregates in solution, particularly at pH values below 8, together with its large fraction of highly mobile residues seriously complicated determination of its three-dimensional structure, both for heteronuclear solution NMR (Grzesiek et al., 1996a, Nat Struct Biol 3:340-345) and for X-ray crystallography (Lee et al., 1996, Cell 85:931-942). Methods used to determine the Nef structure by NMR at pH 8 and 0.6 mM concentration are presented, together with a detailed description of Nef's secondary and tertiary structure. The described techniques have general applicability for the NMR structure determination of proteins that are aggregating and/or have limited stability at low pH values. Extensive chemical shift assignments are reported for backbone and side chain 1H, 13C, and 15N resonances of the HIV-1 Nef deletion mutants NEF delta 2-39, NEF delta 2-39, delta 159-173, and of NEF delta 2-39, delta 159-173 in complex with the SH3 domain of the Hck tyrosine protein kinase. Besides a type II polyproline helix, Nef's structure consists of three alpha-helices, a 3(10) helix, and a five-stranded anti-parallel beta-sheet. The analysis of 15N relaxation parameters of the backbone amide sites reveals that all the secondary structure elements are non-mobile on the picosecond to nanosecond and on the millisecond time scale. A large number of slowly exchanging amide protons provides evidence for the stability of the Nef core even on the time scale of hours. Significant internal motions on the ps to ns time scale are detected for residues 60 to 71 and for residues 149 to 180, which form solvent-exposed loops. The residues of the HIV-1 protease cleavage site (W57/L58) do not exhibit large amplitude motions on the sub-nanosecond time scale, and their side chains insert themselves into a hydrophobic crevice formed between the C-terminus of helix 1 and the N-terminus of helix 2. A refined structure has been determined based on additional constraints for side-chain and backbone dihedral angles derived from a large number of three-bond J-coupling and ROE data.


Cardiovascular effects of (-)-11-OH-delta 8-tetrahydrocannabinol-dimethylheptyl in rats.
The effects of the stereochemically pure psychoactive cannabinoid (-)-11-OH-delta 8-tetrahydrocannabinol-dimethylheptyl (HU-210) on blood pressure (BP) and heart rate (HR) were determined in rats. In pentobarbital-anesthetized animals, the compound produced dose-related, long-lasting hypotension and bradycardia at doses between 10 and 1,000 micrograms/kg. BP began to decrease immediately after drug administration, and in no case was an initial pressor response observed. Previous vagotomy (VX) or pretreatment with 6-hydroxydopamine (6-OHDA) did not affect hypotension. Bradycardia was inhibited by VX, but only 60 min after administration of HU-210; it was enhanced by 6-OHDA. The cannabinoid blocked reflex bradycardia induced by phenylephrine (PE). HU-210 also decreased BP and HR in conscious rats. Hypotension lasted 2 h, whereas bradycardia was still present 8 h after drug administration. HU-210 thus shares with delta 9-tetrahydrocannabinol (THC) the ability to decrease BP and HR, but is 5-10 times more potent than the natural compound. Its lack of an initial pressor effect, such as that described for THC, could be related to its specificity for the type-1 cannabinoid (CB1) receptor. Hypotension and bradycardia after HU-210 administration are not due to sympathetic withdrawal. Enhanced parasympathetic tone is involved in bradycardia only at a late stage of the response.


Insertion of a myc-tag within α-dystroglycan domains improves its biochemical and microscopic detection.
Epitope tags and fluorescent fusion proteins have become indispensable molecular tools for studies in the fields of biochemistry and cell biology. The knowledge collected on the subdomain organization of the two subunits of the adhesion complex dystroglycan (DG) enabled us to insert the 10 amino acids myc-tag at different locations along the α-subunit, in order to better visualize and investigate the DG complex in eukaryotic cells. We have generated two forms of DG polypeptides via the insertion of the myc-tag 1) within a flexible loop (between a.a. 170 and 171) that separates two autonomous subdomains, and 2) within the C-terminal domain in position 500. Their analysis showed that double-tagging (the β-subunit is linked to GFP) does not significantly interfere with the correct processing of the DG precursor (pre-DG) and confirmed that the α-DG N-terminal domain is processed in the cell before α-DG reaches its plasma membrane localization. In addition, myc insertion in position 500, right before the second Ig-like domain of α-DG, proved to be an efficient tool for the detection and pulling-down of glycosylated α-DG molecules targeted at the membrane. Further characterization of these and other myc-permissive site(s) will represent a valid support for the study of the maturation process of pre-DG and could result in the creation of a new class of intrinsic doubly-fluorescent DG molecules that would allow the monitoring of the two DG subunits, or of pre-DG, in cells without the need of antibodies.


Lot-to-Lot Variation.
Lot-to-lot variation affecting calibrators and reagents is a frequent challenge that limits the laboratory's ability to produce consistent results over time. This variation is not without clinical consequence and there are several well-documented examples of adverse clinical outcomes. It is important that laboratories have procedures in place for quantification of this inaccuracy, and for determining whether the amount of variation is acceptable for the release of patient results. Various approaches have been taken to the assessment of new lots, including the evaluation protocol published by the Clinical and Laboratory Standards Institute (CLSI). Internal quality control and external quality assurance material is often not commutable, and so the use of native patient samples is preferred. Published evaluation protocols differ significantly in ease of use and statistical rigour, and some may be underpowered to detect a clinically meaningful change between lots. Furthermore, current protocols (including the CLSI protocol) will not detect cumulative shifts between reagent lots. This shortcoming may at least partly be addressed by laboratories adopting moving patient averages or similar quality procedures. Collaboration and data-sharing between laboratories and manufacturers also has an important role to play in the detection of lot-to-lot variation. While the laboratory may take steps to evaluate and detect variation, the ideal is to reduce variation between lots at the point of manufacture. Using appropriate acceptance criteria based on medical need or biological variation requirements instead of some arbitrary percentage may go some steps toward achieving this.


Identification of human remains from the Second World War mass graves uncovered in Bosnia and Herzegovina.
To present the results obtained in the identification of human remains from World War II found in two mass graves in Ljubuški, Bosnia and Herzegovina. Samples from 10 skeletal remains were collected. Teeth and femoral fragments were collected from 9 skeletons and only a femoral fragment from 1 skeleton. DNA was isolated from bone and teeth samples using an optimized phenol/chloroform DNA extraction procedure. All samples required a pre-extraction decalcification with EDTA and additional post-extraction DNA purification using filter columns. Additionally, DNA from 12 reference samples (buccal swabs from potential living relatives) was extracted using the Qiagen DNA extraction method. QuantifilerTM Human DNA Quantification Kit was used for DNA quantification. PowerPlex ESI kit was used to simultaneously amplify 15 autosomal short tandem repeat (STR) loci, and PowerPlex Y23 was used to amplify 23 Y chromosomal STR loci. Matching probabilities were estimated using a standard statistical approach. A total of 10 samples were processed, 9 teeth and 1 femoral fragment. Nine of 10 samples were profiled using autosomal STR loci, which resulted in useful DNA profiles for 9 skeletal remains. A comparison of established victims' profiles against a reference sample database yielded 6 positive identifications. DNA analysis may efficiently contribute to the identification of remains even seven decades after the end of the World War II. The significant percentage of positively identified remains (60%), even when the number of the examined possible living relatives was relatively small (only 12), proved the importance of cooperation with the members of the local community, who helped to identify the closest missing persons' relatives and collect referent samples from them.


Puerarin inhibits iNOS, COX-2 and CRP expression via suppression of NF-κB activation in LPS-induced RAW264.7 macrophage cells.
Puerarin (7,4'-dihydroxy-8-C-glucosylisoflavone) is the most abundant isoflavone-C-glucoside extracted from Radix puerariae, and it has been used for various medicinal purposes in traditional oriental medicine for thousands of years. In the present study, the ability of the puerarin to modulate inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and C reactive protein (CRP) expression and induce changes in the nuclear factor κB (NF-κB) pathway in RAW264.7 macrophage cells was examined. The protein and mRNA levels of lipopolysaccharide (LPS)-induced iNOS, COX-2 and CRP were determined in RAW246.7 macrophage cells. Inhibitor κB (I-κB) phosphorylation and p65NF-κB expression in RAW246.7 macrophage cells were also detected under our experimental conditions. The results indicated that puerarin inhibited the expression of LPS-induced iNOS, COX-2 and CRP proteins and also suppressed their mRNAs from RT-PCR experiments in RAW264.7 cells. Subsequently, we determined that the inhibition of iNOS, COX-2 and CRP expression was due to a dose-dependent inhibition of phosphorylation and degradation of I-κB, which resulted in the reduction of p65NF-κB nuclear translocation. These data suggested that the effect of puerarin-mediated inhibition of LPS-induced iNOS, COX-2 and CRP expression is attributed to suppressed NF-κB activation at the transcriptional level.


Cognitive-behavioral and pharmacological interventions for premenstrual syndrome or premenstrual dysphoric disorder: a meta-analysis.
The current meta-analysis investigates the efficacy of psychotherapeutic interventions and psychopharmacotherapy for premenstrual syndrome (PMS) and premenstrual dysphoric disorder. Based on a multiple-phase literature search, controlled trials were selected according to a priori defined inclusion criteria. Data were extracted on the basis of a standardized coding scheme. The standardized weighted mean difference (random effects model) was used as effect size index. Dependent on outcome, 22 included studies obtained small to medium effect sizes for cognitive-behavioral interventions (range: d(+) = 0.24-0.70) and for serotonergic antidepressants (range: d(+) = 0.29-0.58), at post-assessment. Follow-ups were performed only in studies of cognitive-behavioral interventions (range: d(+) = 0.46-0.74). There was no evidence of a publication bias. For both cognitive-behavioral interventions and serotonergic antidepressants, efficacy in treatment of PMS was found to not be satisfactory. Future research should possibly focus more on a combination of both approaches.


Depressed smokers and stage of change: implications for treatment interventions.
Tobacco Dependence among smokers with psychiatric disorders has been under-addressed by the mental health, addictions, and tobacco control communities. This study examined depressed smokers' readiness to quit and the applicability of the Stages of Change framework to a psychiatric sample. Currently depressed smokers (N=322) were recruited from four outpatient psychiatric clinics. Participants averaged 16 cigarettes per day (S.D.=10) and 24 years (S.D.=13) of smoking. The majority (79%) reported intention to quit smoking with 24% ready to take action in the next 30 days. Individuals in the preparation stage reported more prior quit attempts, a greater commitment to abstinence, increased recognition of the cons of smoking, and greater use of the processes of change. Precontemplators were least likely to identify a goal related to their smoking behavior. Depressive symptom severity and history of recurrent depressive episodes were unrelated to readiness to quit. This study is one of the first to examine the smoking behaviors of currently depressed psychiatric outpatients. The level and longevity of their tobacco use underscore the need for cessation interventions. The consistency in hypothesized patterns among theoretical constructs of the Stages of Change model supports the transfer of stage-tailored interventions to this clinical population.


Mutations in an integration host factor-binding site: effect on lambda site-specific recombination and regulatory implications.
The manner in which integration host factor (IHF) regulates lambda site-specific recombination has been analyzed by examining the behavior of both wild-type and mutant DNAs in integrative and excisive recombination as well as in protein binding. While integrative recombination of an attP with two base changes in the H1 site required 8-fold more IHF than did wild type, binding to this site was lowered at least 500-fold, suggestive of cooperative interactions. A mutant attP with nine base changes did not integrate at all in vitro, with the defect being less severe in vivo. IHF inhibition of excisive recombination was relieved by both mutations in vitro and in vivo. These results imply that occupancy of the H1 site is critical for determining the direction of recombination. It is proposed that IHF inhibition of excision provides a monitor of the strength of the induction stimulus and the nutritional state of the cell; this would allow the prophage to excise selectively in conditions which favor successful completion of the lytic cycle.


DNA methylome analysis of acute lymphoblastic leukemia cells reveals stochastic de novo DNA methylation in CpG islands.
To identify regions of aberrant DNA methylation in acute lymphoblastic leukemia (ALL) cells of different subtypes on a genome-wide scale. Whole-genome bisulfite sequencing (WGBS) was used to determine the DNA methylation levels in cells from four pediatric ALL patients of different subtypes. The findings were confirmed by 450k DNA methylation arrays in a large patient set. Compared with mature B or T cells WGBS detected on average 82,000 differentially methylated regions per patient. Differentially methylated regions are enriched to CpG poor regions, active enhancers and transcriptional start sites. We also identified approximately 8000 CpG islands with variable intermediate DNA methylation that seems to occur as a result of stochastic de novo methylation. WGBS provides an unbiased view and novel insights into the DNA methylome of ALL cells.


Etiology of pneumonia in children in the absence of pneumococcal and antihaemophilus vaccines.
Childhood pneumonia represents an important pathology, a cause of morbidity and mortality worldwide. Our study aims to determine etiology of pneumonia in hospitalized children using several laboratory methods. We performed a prospective study that enrolled 560 children age 1 up to 18 years old all diagnosed with pneumonia by clinical and radiological features. We applied various laboratory methods (serologic, bacteriologic: bronchial aspirate, sputum, pleural effusion and blood culture) in order to identify a pathogen agent that caused pneumonia. Statistics used Statistical Package for Social Science. An etiology was established in 68.92% of all cases included in the study, as follows: in 33.93% viral etiology, in 25.13% we identified Streptococcus pneumoniae, in 20.2% Mycoplasma pneumoniae, Klebsiella pneumoniae in 8.29%, Staphylococcus aureus in 7.51%, Haemophilus influenzae in 4.92%. Mixed bacterial and viral infection was identified in 4.40% of all cases. A potential causative agent of childhood pneumonia was determined in most cases, S. pneumoniae being the main agent involved in community acquired childhood pneumonia in our country.


Differential effect of impaired renal function on the kinetics of clavulanic acid and amoxicillin.
Amoxicillin and clavulanic acid are prescribed as a fixed drug combination. The purpose of the present study was to assess the influence of various degrees of renal insufficiency (glomerular filtration rate [GFR], less than 5 to greater than 75 ml/min per 1.73 m2) on the pharmacokinetics of amoxicillin and clavulanic acid following oral (500 and 125 mg of amoxicillin and clavulanic acid, respectively) and intravenous (1,000 and 200 mg, respectively) dosing. The volume of distribution and the systemic availability were independent of the renal function, while the total body clearance and the renal and the nonrenal clearance of amoxicillin and clavulanic acid decreased with decreasing renal function. The decrease in the total body clearance was more pronounced for amoxicillin than for clavulanic acid. This explains the increase in the ratio of the area under the plasma concentration versus time curve of amoxicillin to that of clavulanic acid with decreasing glomerular filtration rate after oral dosing; for example for a GFR of 75 ml/min, the ratio of amoxicillin to clavulanic acid was 4.9 +/- 1.2; for a GFR of 35 to 75 ml/min, 5.3 +/- 2.4; for a GFR of 10 to 35 ml/min, 11.9 +/- 5.8; for a GFR of 5 to 10 ml/min, 13.4 +/- 9.1; and for patients on hemodialysis, 14.7 +/- 5.3. Dosage recommendations are suggested which prevent undue accumulations of amoxicillin while maintaining adequate concentrations of clavulanic acid.


Meta-analysis of prostate-specific antigen and digital rectal examination as screening tests for prostate carcinoma.
Physicians commonly screen for prostate cancer by using prostate-specific antigen (PSA) and digital rectal examination (DRE). The usefulness of these screening mechanisms is not well established, however. A meta-analysis of PSA and DRE to detect prostate carcinoma was conducted with a focus on sensitivity, specificity, and positive predictive value. A literature search of OVID database (1966 to November 1999) using the medical subject headings "prostate-specific antigen" and "mass screening," as well as "prostate carcinoma," was performed. Thirteen articles were selected for the meta-analysis in this study. Most studies included asymptomatic men older than 50 years from various countries. Pooled results were calculated from the individual reports for sensitivity, specificity, and positive predictive value for PSA and DRE based on biopsy result as the reference standard. The overall detection rate of prostate carcinoma was 1.8% based on a positive biopsy. Of the prostate carcinoma detected, 83.4% was localized. The pooled sensitivity, specificity, and positive predictive value for PSA were 72.1%, 93.2% and 25.1%, respectively; and for DRE were 53.2%, 83.6% and 17.8%, respectively. There were two major outcomes of this meta-analysis. One was the potential for detecting early-stage prostate cancer with these screening tests, because 83.4% of total cancers detected were localized. The second important outcome was that the overall sensitivity, specificity, and positive predictive value for PSA were higher than those for DRE when used as a screening tool to detect prostate cancer. When a patient has abnormal findings using PSA and DRE, the chance of cancer is 1 in 4 or 5. Conversely, when findings from PSA and DRE are normal, the chance of missing a cancer is about 10%.


GNTD: reconstructing spatial transcriptomes with graph-guided neural tensor decomposition informed by spatial and functional relations.
Spatially-resolved RNA profiling has now been widely used to understand cells' structural organizations and functional roles in tissues, yet it is challenging to reconstruct the whole spatial transcriptomes due to various inherent technical limitations in tissue section preparation and RNA capture and fixation in the application of the spatial RNA profiling technologies. Here, we introduce a graph-guided neural tensor decomposition (GNTD) model for reconstructing whole spatial transcriptomes in tissues. GNTD employs a hierarchical tensor structure and formulation to explicitly model the high-order spatial gene expression data with a hierarchical nonlinear decomposition in a three-layer neural network, enhanced by spatial relations among the capture spots and gene functional relations for accurate reconstruction from highly sparse spatial profiling data. Extensive experiments on 22 Visium spatial transcriptomics datasets and 3 high-resolution Stereo-seq datasets as well as simulation data demonstrate that GNTD consistently improves the imputation accuracy in cross-validations driven by nonlinear tensor decomposition and incorporation of spatial and functional information, and confirm that the imputed spatial transcriptomes provide a more complete gene expression landscape for downstream analyses of cell/spot clustering for tissue segmentation, and spatial gene expression clustering and visualizations.


Psychomimetic reactions after low-dose ketamine infusion. Comparison with neuroleptanaesthesia.
Low-dose ketamine anaesthesia was compared with neuroleptanaesthesia, in respect of immediate and longer-term psychomimetic reactions, in 40 female patients undergoing elective gynaecological surgery. Qualitatively, but not quantitatively, different psychomimetic reactions occurred in both groups. In the neurolept group the predominant complaint was of a dissociative nature, reported after 24 h by 11 (55%) of the patients, while in the ketamine group the predominant experience was dreaming, reported by eight (40%) of the patients after 24 h. Interviews after 3 months revealed a low frequency of psychomimetic reactions in both groups. However, 30% of all the patients (12) complained of impairment of intellectual function, and in seven patients this was severe enough to interfere with their ability to work.


Morphology and functional roles of synoviocytes in the joint.
The joint capsule exhibits a unique cellular lining in the luminal surface of the synovial membrane. The synovial intimal cells, termed synoviocytes, are believed to be responsible for the production of synovial fluid components, for absorption from the joint cavity, and for blood/synovial fluid exchanges, but their detailed structure and function as well as pathological changes remain unclear. Two types of synoviocytes, macrophagic cells (type A cells) and fibroblast-like cells (type B cells) have been identified. Type A synoviocytes are non-fixed cells that can phagocytose actively cell debris and wastes in the joint cavity, and possess an antigen-presenting ability. These type A cells, derived from blood-borne mononuclear cells, can be considered resident macrophages (tissue macrophages) like hepatic Kupffer cells. Type B synoviocytes are characterized by the rich existence of rough endoplasmic reticulum, and dendritic processes which form a regular network in the luminal surface of the synovial membrane. Their complex three-dimensional architecture was first revealed by our recent scanning electron microscopy of macerated samples. The type B cells, which are proper synoviocytes, are involved in production of specialized matrix constituents including hyaluronan, collagens and fibronectin for the intimal interstitium and synovial fluid. The proliferative potentials of type B cells in loco are much higher than type A cells, although the transformation of subintimal fibroblasts into type B cells can not be excluded. In some mammals, type B cells show features suggesting endocrine and sensory functions, but these are not recognized in other species. The synoviocytes, which form a discontinuous cell layer, develop both fragmented basement membranes around the cells and junctional apparatus such as desmosomes and gap junctions. For an exact understanding of the mechanism of arthritis, we need to establish the morphological background of synoviocytes as well as their functions under normal conditions.


Spectral phase transitions in optical parametric oscillators.
Driven nonlinear resonators provide a fertile ground for phenomena related to phase transitions far from equilibrium, which can open opportunities unattainable in their linear counterparts. Here, we show that optical parametric oscillators (OPOs) can undergo second-order phase transitions in the spectral domain between degenerate and non-degenerate regimes. This abrupt change in the spectral response follows a square-root dependence around the critical point, exhibiting high sensitivity to parameter variation akin to systems around an exceptional point. We experimentally demonstrate such a phase transition in a quadratic OPO. We show that the divergent susceptibility of the critical point is accompanied by spontaneous symmetry breaking and distinct phase noise properties in the two regimes, indicating the importance of a beyond nonlinear bifurcation interpretation. We also predict the occurrence of first-order spectral phase transitions in coupled OPOs. Our results on non-equilibrium spectral behaviors can be utilized for enhanced sensing, advanced computing, and quantum information processing.


Inhibition of RNA transportation induces glioma cell apoptosis via downregulation of RanGAP1 expression.
The prognosis of glioblastoma remains poor, even treatment with surgery, radiation, or chemotherapy. Therefore, it is still important to develop a new strategy for treatment of glioblastoma. Previous reports demonstrated that rRNA is produced at abnormally high levels in tumor cells. Nuclear export of all non-coding RNAs are known to depend on RanGTPase system. Hydrolyzation of RanGTP-RNA complex by RanGTPase activating protein 1 (RanGAP1) releases RNA from nucleus to cytoplasm. Therefore, inhibition of RNA transportation would be a useful strategy to affect cancer cell fate. In this study, 5-30 μM of oridonin, a natural diterpenoid compound isolated from the traditional Chinese medicine, Rabdosia rubescens, induced U87MG glioma cell apoptosis and RNA accumulation in nucleus at 12h-time point. Before U87MG cell apoptosis, the RanGAP1 protein amount decreased and RanGTP accumulated in nucleus as respectively determined by immunoprecipitation and immunofluorescence, suggesting that decrease of RanGAP1 may result in nuclear entrapment of RanGTP and RNA, and then induce U87MG cell death. In contrast, over-expression of the RanGAP1 protein reversed oridonin-induced U87MG cell apoptosis. Hence, we demonstrated that downregulation of the RanGAP1 protein level by oridonin may result in RNA accumulation in nucleus via nuclear entrapment of RanGTP which eventually led to the apoptosis of glioma cells.


Promiscuous hydroxylases for the functionalization of polycyclic tetramate macrolactams--conversion of ikarugamycin to butremycin.
Polycyclic tetramate macrolactams (PTMs) are a structurally, biomedically and biosynthetically intriguing class of bacterial metabolites. By combining parts of the machineries of different PTM biosynthetic pathways, we demonstrate for the first time the substrate promiscuity of a class of PTM tailoring enzymes, thereby facilitating the (bio)synthesis of butremycin.