Expanded endonasal approach: a fully endoscopic completely transnasal resection of a skull base arteriovenous malformation.
Vascular lesions with an intraosseus nidus involving the skull base are uncommon and challenging [Gianoli GJ, Amedee RG Vascular malformation of the sphenoid sinus. Ear Nose Throat J. 70:373-375;(1991), Malik GM, Mahmood A, Mehta BA Dural arteriovenous malformation of the skull base with intraosseous vascular nidus. Report of two cases. J. Neurosurg 81:620-623;(1994)]. We present a pediatric patient, with a life-threatening arteriovenous malformation (AVM) of the sphenoid sinus, clivus, and ventral skull base, who failed routine multimodality management of AVMs. An entirely transsphenoidal fully endoscopic resection was used to resect this ventral cranial base AVM with an intraosseus nidus located in the clivus. A 4-year-old female presented with recurrent, life-threatening hemorrhages from a clival and ventral skull base AVM of the entire clivus and ventral skull base. The patient had been temporized from the age of 2-4 years with multiple internal and external carotid arterial particulate and alcohol embolizations, including both external and internal carotid artery embolizations, intracranial ligation of the right internal carotid artery, and gamma knife irradiation. Despite these multiple interventions, the patient had persistent, life-threatening hemorrhages from arterial recanalization and recruitment requiring intubation, tracheostomy, and nasopharyngeal packing. The patient underwent a three-stage surgical intervention to resect the AVM. An open subfrontal approach, as the first procedure, provided minimal access to the feeding vessels and was therefore aborted. A two-stage image-guided fully endoscopic approach via a sublabial midface approach without external incisions was performed. Postoperative angiography revealed minimal residual shunting in the pharynx and cavernous sinus. The patient has been free of significant hemorrhages over the past three years. Technological advances in endoscopic surgery and image guidance are now allowing for purely endoscopic surgical treatment of previously unresectable lesions with acceptable morbidity. We report the successful and safe resection of a ventral cranial base AVM via a fully endoscopic approach. This paper reports the first AVM with a purely intraosseus nidus of the ventral skull base and demonstrates the ability to deal with complex ventral skull base lesions using a fully endoscopic transsphenoidal technique.


Estimated percentile curves of occluding pairs in an adult population.
The purpose of this study was to provide the estimated percentile curves of occlusal pairs (OPs) in adults for use in oral health instruction. Data on number of present teeth (PT) were obtained from the National Report on the Survey of Dental Disease in 2005. Data from 1,535 males and 2,248 females, ranging in age from 18 to 82, were analysed. The mean number of OPs were used from our previous report. In that report we can be estimated OPs from the number of PT. In this study, PT data were replaced by the mean number of OPs. Estimated percentile curves in males, the 10th percentile of OPs was 13.7 at 60 years of age and 12.5 at 70. The 50th percentile of OPs was 12.2 at 50 years of age, 11.2 at 60 and 5.4 at 70. The 90th percentile of OPs was 7.6 at 50 years of age, 2.1 at 60 and 0 at 70. There appeared to be a cut off point after 10 OPs where the 50th to 97th percentile curves of OPs dropped off rapidly. In females, the 10th percentile of OPs was 13.4 at 60 years of age and 12.7 at 70. The 50th percentile of OPs was 11.5 at 50 years of age, 10.2 at 60 and 6.2 at 70. The 90th percentile of OPs was 7.1 at 50 years of age, 2.4 at 60 and 0 at 70. In females again the percentile curves dropped off after 10 OPs. Our results suggest that the loss of OPs may be accelerated when the number of OPs drops below 10. This means that patients need at least 10 OPs in order to maintain a stable oral status. The estimated percentile curves given in this study should be made available for use in adult oral health education.


Building clinical capacity for fetal alcohol spectrum disorder diagnoses in western and northern Canada.
Fetal alcohol syndrome and fetal alcohol spectrum disorder are common problems. In response to this problem the Canada Northwest FASD Research Network was established in 2005 by the Canada Northwest FASD Ministerial Partnership. This study was conducted to determine the FASD clinical activity in Canada Northwest. The Network identified all clinical programs via Internet sites, provincial postings and professional word of mouth references that purported to do FASD assessments regularly using a multidisciplinary assessment team. Each of these programs was sent a questionnaire asking about clinical capacity, aggregate diagnostic results, team composition, time of clinical assessment and cost of assessment. Of the 27 programs identified to receive the questionnaire 15 programs responded. These programs were determined to have evaluated about 85% of the patients evaluated by all the programs. The total 7 jurisdictional capacity for FASD diagnosis was 816 evaluations in 2005 and projected to be 975 in 2006. Selection methods for appointing patients for assessment seemed excellent as 23% of those assessed were found to have FAS or pFAS and another 44% had other forms of FASD. The most common professionals to participate in the team evaluations were Paediatricians, Clinical Psychologists, Speech and Language Pathologists and Occupational Therapists. Clinics are developing in western and northern Canada to diagnose patients with FASD. Comparing the experiences of these clinics can help to determine the continued need to increase diagnostic capacity, standardize diagnostic approaches to assure consistency of approach and diagnosis across the sites and appropriately staff and fund the programs. Key words: FASD; diagnosis; Canada; clinics.


Use and diversion of medical marijuana among adults admitted to inpatient psychiatry.
Marijuana use is associated with anxiety, depressive, psychotic, neurocognitive, and substance use disorders. Many US states are legalizing marijuana for medical uses. To determine the prevalence of medical marijuana use and diversion among psychiatric inpatients in Colorado. Some 623 participants (54.6% male) responded to an anonymous 15-item discharge survey that assessed age, gender, marijuana use, possession of a medical marijuana card, diversion of medical marijuana, perceived substance use problems, and effects of marijuana use. Univariate statistics were used to characterize participants and their responses. Chi-square tests assessed factors associated with medical marijuana registration. Of the total number of respondents, 282 (47.6%) reported using marijuana in the last 12 months and 60 (15.1%) reported having a marijuana card. In comparison to survey respondents who denied having a medical marijuana card, those respondents with a medical marijuana card were more likely to have initiated use before the age of 25, to be male, to have used marijuana in the last 12 months, and to have used at least 20 days in the past month. 133 (24.1%) respondents reported that someone with a medical marijuana card had shared or sold medical marijuana to them; 24 (41.4%) of respondents with a medical marijuana card reported ever having shared or sold their medical marijuana. Medical marijuana use is much more prevalent among adults hospitalized with a psychiatric emergency than in the general population; diversion is common. Further studies which correlate amount, dose, duration, and strain of use with particular psychiatric disorders are needed.


Diversity and Genetic Structure of Theileria annulata in Pakistan and Other Endemic Sites.
Theileria annulata is a tick-borne protozoan parasite responsible for bovine theileriosis, a disease that impacts cattle population in many developing countries. Development and deployment of effective control strategies, based on vaccine or therapy, should consider the extent of diversity of the parasite and its population structure in different endemic areas. In this study, we examined T. annulata in Pakistan and carried out a comparative analysis with similar data garneted in other areas, to provide further information on the level of parasite diversity and parasite genetic structure in different endemic areas. The present study examined a set of 10 microsatellites/minisatellites and analyzed the genetic structure of T. annulata in cattle breeds from Pakistan (Indian sub-continent) and compared these with those in Oman (Middle East), Tunisia (Africa), and Turkey (Europe). A high level of genetic diversity was observed among T. annulata detected in cattle from Pakistan, comparable to that in Oman, Tunisia, and Turkey. The genotypes of T. annulata in these four countries form genetically distinct groups that are geographically sub-structured. The T. annulata population in Oman overlapped with that in the Indian Subcontinent (Pakistan) and that in Africa (Tunisia). The T. annulata parasite in Pakistan is highly diverse, and genetically differentiated. This pattern accords well and complements that seen among T. annulata representing the global endemic site. The parasite population in the Arabian Peninsula overlapped with that in the Indian-Subcontinent (India) and that in Africa (Tunisia), which shared some genotypes with that in the Near East and Europe (Turkey). This suggests some level of parasite gene flow, indicative of limited movement between neighboring countries.


Scottish country dance: benefits to functional ability in older women.
The effects of long-term participation in Scottish country dance on body composition, functional ability, and balance in healthy older females were examined. Participants were grouped into dancers and physically active nondancers (ages 60-70 and 70-80 for both groups). Physical activity, body composition (body-mass index, skinfold thickness, waist-to-hip ratio), functional ability (6-min walk distance, 6-m walk time, 8-ft up-and-go time, lower body flexibility, shoulder flexibility), and static balance were measured. Younger dancers and physically active nondancers had similar 6-min walk distance, 6-m walk time, and 8-ft up-and-go time results; however, while older dancers performed similarly to younger dancers, older physically active nondancers performed poorer than their younger counterparts (p < .05). Body composition and static balance were the same for all groups. Regular physical activity can maintain body composition and postural stability with advancing age; however, Scottish country dance can delay the effects of aging on locomotion-related functional abilities.


Mean platelet volume and other platelet volume indices in patients with stable coronary artery disease and acute myocardial infarction: A case control study.
Coronary artery disease is mainly caused by atherosclerosis and its complications. Platelets and their activity have an important role in initiation of atherosclerotic lesions and coronary thrombus formation. Larger platelets are enzymatically and metabolically more active and have a higher potential thrombotic ability as compared with smaller platelets. To study the changes in platelet volume indices and platelet count in acute myocardial infarction, stable coronary artery disease and compare them with controls to assess their usefulness in predicting coronary events. This was a comparative study of 128 subjects; 39 patients with acute myocardial infarction (AMI), 24 patients with stable coronary artery disease (SCAD) and 65 controls. Venous sample were drawn from AMI subjects on admission (within 4 hours of chest pain) and collected in standardized EDTA sample tubes. Platelet count and volume indices were assayed within 30 minutes of blood collection, using Sysmex KX21-N autoanalyzer. Venous samples were also drawn from SCAD on who were admitted for angiography and subject attending routine checkups. The mean platelet volume was significantly higher in patients with AMI (9.65 ± 0.96) as compared to SCAD (9.37 ± 0.88) and controls (9.21 ± 0.58). The best cut-off values for MPV when predicting AMI and SCAD in patients were 9.25 fl (sensitivity 56.4%; specificity 45.9%) and 9.15 fl (sensitivity 54.2%; specificity 42.23%), respectively. Measurements of MPV may be of some benefit in detecting those patients at higher risk for an AMI and CAD.


Deletion of antigen-specific immature thymocytes by dendritic cells requires LFA-1/ICAM interactions.
An in vitro assay was used for assessing the participation of various cell surface molecules and the efficacy of various cell types in the deletion of Ag-specific immature thymocytes. Thymocytes from mice expressing a transgenic TCR specific for the male Ag presented by the H-2Db class I MHC molecule were used as a target for deletion. In H-2d transgenic mice, cells bearing the transgenic TCR are not subjected to thymic selection as a consequence of the absence of the restricting H-2Db molecule but, nevertheless, express this TCR on the vast majority of immature CD4+8+ thymocytes. In this report we show that CD4+8+ thymocytes from H-2d TCR-transgenic mice are preferentially killed upon in vitro culture with male APC; DC were particularly effective in mediating in vitro deletion when compared with either B cells or T cells. Deletion of CD4+8+ thymocytes by DC was H-2b restricted and could be inhibited by mAb to either LFA-1 alpha or CD8. Partial inhibition was observed with mAb to ICAM-1, whereas mAb to CD4 and LFA-1 beta were without effect. These results are the first direct evidence of LFA-1 involvement in negative selection and provide further direct support for the participation of CD8/class I MHC interactions in this process. Like the requirements for deletion, activation of mature male-specific CD4-8+ T cells from female H-2b TCR-transgenic mice was also largely dependent on Ag presentation by DC and required both LFA-1/ICAM and CD8/class I MHC interactions; these results support the view that activation and deletion may represent maturation stage-dependent consequences of T cells encountering the same APC. Finally, our results also support the hypothesis that negative selection (deletion) does not require previous positive selection because deletion was observed under conditions where positive selection had not occurred.


[Preoperative evaluation--an update].
The length of hospital stay shortens increasingly. Even more patients are assessed and clarified in the outpatient clinic. Preoperative evaluation is performed ambulatory or by the general practitioner himself. There is still a trend towards fewer and more selective examinations. The value of evaluation algorithms is subject of debate nowadays. Moreover, these extensive and abstract procedures are not always helpful in clinical practice. The perioperative cardioprotection with beta-blockers and statins and the continuation of antiplatelet therapy seems to have significance in reduction of perioperative cardiac events. However, a careful physical examination and a precise medical history of the patient have not become less important today. They contribute significantly to an efficient preoperative evaluation.


A national register for long-term oxygen therapy in chronic hypoxia: preliminary results.
A national register of patients undergoing long-term oxygen therapy in Sweden was started in 1987. Of the population of 8.4 million, 560 patients (267 males) were registered as undergoing domiciliary oxygen treatment on January 1, 1987. These registered patients, aged between 2-86 yrs (mean age 65 yrs), constituted some 90% of all patients receiving domiciliary oxygen therapy because of chronic hypoxaemia on that date. The chronic respiratory diseases leading to hypoxaemia (more than one diagnosis could be registered for each patient) were: chronic obstructive pulmonary disease (COPD) (393), sequelae from pulmonary tuberculosis (92), thoracic deformity (97), interstitial fibrosis (44), benign pleural disease (22) and others (84). Concentrators were used by 253 patients and high-pressure compressed gas cylinders by 307. Arterial blood gas analyses were registered for 547 patients. The mean arterial oxygen tension (PaO2) when breathing air was 6.5 +/- 1.1 kPa and 9.0 +/- 1.4 kPa when breathing oxygen. The mean arterial carbon dioxide tension (PaCO2) without oxygen was 6.5 +/- 1.5 kPa and 6.8 +/- 1.5 kPa with oxygen. The register forms a data-base which can be used for the evaluation of different home oxygen systems, regional differences in the access to treatment and treatment performance and decisions relating to health care economics.


BMP-regulated exosomes from Drosophila male reproductive glands reprogram female behavior.
Male reproductive glands secrete signals into seminal fluid to facilitate reproductive success. In Drosophila melanogaster, these signals are generated by a variety of seminal peptides, many produced by the accessory glands (AGs). One epithelial cell type in the adult male AGs, the secondary cell (SC), grows selectively in response to bone morphogenetic protein (BMP) signaling. This signaling is involved in blocking the rapid remating of mated females, which contributes to the reproductive advantage of the first male to mate. In this paper, we show that SCs secrete exosomes, membrane-bound vesicles generated inside late endosomal multivesicular bodies (MVBs). After mating, exosomes fuse with sperm (as also seen in vitro for human prostate-derived exosomes and sperm) and interact with female reproductive tract epithelia. Exosome release was required to inhibit female remating behavior, suggesting that exosomes are downstream effectors of BMP signaling. Indeed, when BMP signaling was reduced in SCs, vesicles were still formed in MVBs but not secreted as exosomes. These results demonstrate a new function for the MVB-exosome pathway in the reproductive tract that appears to be conserved across evolution.


Dynamics and stability of the metal binding domains of the Menkes ATPase and their interaction with metallochaperone HAH1.
Human copper-ATPases ATP7A and ATP7B are essential for intracellular copper homeostasis. The main roles of the Menkes protein, ATP7A, are the delivery of copper to the secretory pathway and the export of excess copper from the enterocytes. The N-terminal domain of membrane protein ATP7A consists of six repetitive sequences of 60-70 amino acids (Mnk1-Mnk6) that fold into individual metal binding domains (MBDs) and bind a single copper ion in the reduced Cu(I) form via two cysteine residues. The structure of each individual MBD is known from nuclear magnetic resonance experiments. Here, we were interested in the stability and dynamics of each isolated MBD in their apo and holo forms and their interactions with the soluble metallochaperone HAH1 that delivers copper to ATP7A. Using molecular dynamics simulations of the MBDs under different conditions, we show that some MBDs (Mnk1 and Mnk5) present large root-mean-square deviations from initial structures or large root-mean-square fluctuations, and great care has to be taken in setting up the simulations. We propose that the first MBD, Mnk1, probably important in the transfer of copper between the metallochaperone and ATPase, could be stabilized by interactions with other MBDs, including a domain located in the loop between Mnk1 and Mnk2. An important result of this work is the apparent direct correlation between the difference in the fluctuations of the metal binding site loop in its apo and holo forms and the measured affinity of the MBD for copper. This difference decreases from Mnk1 to Mnk6, Mnk4, and Mnk2 in this order. The study of the exposure to the solvent of the metal and the residues of the metal binding loop of the MBDs also shows different behavior for each MBD. In particular, copper in serine-rich domain Mnk3 and largely fluctuating domain Mnk5 appears to be more solvent-exposed than in the other MBDs. In the second part of this work, we investigated the importance of electrostatics in the MBD-chaperone interactions using different docking programs. Mnk1 and Mnk4 present a large electrostatic dipole moment and large stabilizing interaction energies with HAH1. Finally, we propose a model structure of ATP7A from Mnk6 (E561) to P1413 based on the crystal structure of LpCopA and docking simulations.


HIV, ethnicity and travel: HIV infection in Vietnamese Australians associated with injecting drug use.
The movement of people with their constructed identities including ethnicity has always been one of the determinants of the human immunodeficiency virus (HIV) pandemic. An example of the contributions of travel and ethnicity to experiences of HIV can be seen in the Vietnamese community in Australia. This paper seeks to describe the contributions of ethnicity and travel to the Australian HIV epidemic with particular reference to the evolving epidemic within the Vietnamese Australian community. We reviewed the available data on the HIV epidemic in Australia with reference to overseas acquisition, ethnicity, the epidemic in the Vietnamese community and the determinants of the current patterns of transmission within this community. Available data suggests that 20-25% of HIV infections notified in Australia are acquired overseas. This proportion is higher in some specific categories such as heterosexually acquired infections. Notification rates are no higher in Vietnamese Australians than in the general Australian population apart from infections associated with injecting drug use (IDU) notified in the state of Victoria. The reasons for this increased rate of notification include increased vulnerability to blood borne virus infection in Australia and the additional, unique risk of frequent travel to Vietnam, a country where IDU carries a high risk of HIV infection. Australia has succeeded in stabilising the HIV epidemic partly through successful interventions to limit the spread of infection among IDUs. There is now early evidence that HIV transmission may be increasing amongst Vietnamese Australian IDUs. Timely responses that help Vietnamese Australian IDUs reduce their accumulation of risk are likely to be important in determining the level of harm associated with IDU throughout Australia.


Inappropriate expression of blood group antigens in hepatic allografts.
We examined the expression of blood group antigens of the ABO, Lewis and Kell antigen systems using monoclonal antibodies and immunohistochemical study on 42 liver allograft specimens from 33 patients who underwent liver transplantation between 1986 and 1991 to learn whether altered blood group antigen expression might have a bearing on the immunopathogenesis of transplant rejection. Specimens were obtained at intervals of 0 days to 3 yr after transplant; they yielded the following histological diagnoses; time zero (n = 4), acute rejection (n = 4), pure cholestasis (n = 4), biliary obstruction (n = 4), early chronic rejection (n = 4), end-stage chronic rejection (n = 15) and miscellaneous late posttransplant biopsies (n = 7). Aberrant expression of blood group antigens was observed in 5 of 15 patients with chronic rejection. Two transplants into the same group O patient showed aberrant expression of AB antigens on hepatocytes, with a canalicular pattern, in group O-transplanted livers. In all three cases in which a group O liver was transplanted into a group A recipient and histological signs of chronic rejection were present, antibody staining showed acquisition of recipient blood phenotype by the donor liver bile ducts, endothelium or both. Aberrant expression of ABO antigens was seen only in chronic rejection. In seven cases we noted canalicular staining of periportal hepatocytes with the Lewis antibodies, normally confined to ducts and ductules. This was associated with severe cholestasis in six of the seven cases and may have represented early ductular metaplasia. These changes in carbohydrate cell surface phenotype may play a role in regulation of hepatic allograft susceptibility to immune-mediated damage.


A single-center real-life study on the use of medical cannabis in patients with dystonia.
While cannabis-based medicine is being commonly used in patients with movement disorders, there is a scarcity of publications regarding the effect of cannabis on dystonia. We aimed to describe medical cannabis use in patients with dystonia and related pain. We employed a structured interview to obtain data on the cannabis treatment regimen, perception of effectiveness and side effect profile. Eligible participants were patients diagnosed with dystonia from the movement disorders unit at the Tel-Aviv Medical Center who had used licensed medical cannabis between January 2019 and January 2021. Twenty-three subjects were interviewed (11 women, mean age 52.7). The most common way of administration was smoking (n = 11). Following an average of 2.5 ± 2.9 years of use, those with widespread dystonia (generalized, hemi and multifocal, n = 11) self-reported on a numeric rating scale an average 63% (range 0%-100%) reduction in symptoms of dystonia, while those with more focal dystonia patterns reported a significantly lower treatment effect of 32%. Participants reported a positive impact in related pain and quality of life, with an average rating of 3.8 out of 5 (SD = 1.2, median = 4) and 3.6 out of 5 (SD = 1.15, median = 4), respectively. Most common side effects were dry mouth (65%), sedation (43%), dizziness (39%) and psychiatric disorders (26%). Three patients (13%) discontinued therapy. A subset of dystonia patients who use medical cannabis under clinical observation reported significant subjective improvement during 30 months of use in average. Further prospective randomized controlled trials are required to examine the effectiveness of cannabis in dystonia.


Assessing disparities in adult vaccination using multimodal approaches in primary care offices: methodology.
Racial disparities in invasive pneumococcal disease and pneumococcal polysaccharide vaccination (PPV) persist despite significant progress. One reason may be that minority patients receive primary care at practices with fewer resources, less efficient office systems, and different priorities. The purposes of this paper are: (1) to describe the recruitment of a diverse array of primary care practices in Pittsburgh, Pennsylvania serving white and minority patient populations, and the multimodal data collection process that included surveys of key office personnel, observations of practice operations and medical record reviews for determining PPV vaccination rates; and (2) to report the results of the sampling strategy. During 2005, 18 practices participated in the study, six with a predominantly minority patient population, nine with a predominantly white patient population, and three with a racial distribution similar to that of this locality. Eight were solo practices and 10 were multiprovider practices; they included federally qualified health centers, privately owned practices and faculty and University of Pittsburgh Medical Center community practices. Providers represented several racial and ethnic groups, as did office staffs. PPV rates determined from 2,314 patients' medical records averaged 60.3 +/- 22.6% and ranged from 11% to 97%. Recruitment of practices with attention to location, patient demographics, and provider types results in a diverse sample of practices and patients. Multimodal data collection from these practices should provide a rich data source for examining the complex interplay of factors affecting immunization disparities among older adults.


Refractive index sensing utilizing a cw photonic crystal nanolaser and its array configuration.
We demonstrate refractive index sensing using photonic crystal point shift nanolasers. These lasers operate continuously at room temperature by photopumping in a liquid, and exhibit a 50-dB peak intensity over the background level and a spectral linewidth of < 26 pm, the resolution limit of the present experiment. The lasing wavelength shifts by soaking in different liquids; the wavelength to index sensitivity was 350 nm/RIU, the highest value recorded to date for nanocavity- based sensors. An index resolution of 9.0 x 10(-5) was thus confirmed, leading to an expectation of a resolution of < 10(-6). We propose and demonstrate a spectrometer-free sensor based on nanolasers in an array configuration. These will be disposable sensors with very simple optical I/O. They are anticipated to be integrated with biochips and used for label-free single molecule detection.


A novel pineal night-specific ATPase encoded by the Wilson disease gene.
We have identified a pineal night-specific ATPase (PINA), a novel splice variant of the ATP7B gene disrupted in Wilson disease (WD). PINA expression exhibits a dramatic diurnal rhythm in both pineal gland and retina with 100-fold greater expression at night than at day. PINA is expressed in pinealocytes and a subset of photoreceptors in adult rats and is transiently expressed in the retinal pigment epithelium and the ciliary body during retinal development. Nocturnal pineal expression of PINA is under the control of a suprachiasmatic nucleus clock mediated by superior cervical ganglion innervation of the pineal. In vitro, PINA expression in pineal cells can be stimulated by agents activating the cAMP signal transduction pathway. PINA is able to restore copper transport activity in Saccharomyces cerevisiae deficient in the homologous copper-transporting ATPase CCC2, suggesting that this protein may function as a copper transporter in rat pinealocytes. These studies suggest a potential role of rhythmic copper metabolism in pineal and/or retina circadian function.


The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosis.
Destruction of the pulmonary epithelium is a major feature of lung diseases caused by the mould pathogen Aspergillus fumigatus. Although it is widely postulated that tissue invasion is governed by fungal proteases, A. fumigatus mutants lacking individual or multiple enzymes remain fully invasive, suggesting a concomitant requirement for other pathogenic activities during host invasion. In this study we discovered, and exploited, a novel, tissue non-invasive, phenotype in A. fumigatus mutants lacking the pH-responsive transcription factor PacC. Our study revealed a novel mode of epithelial entry, occurring in a cell wall-dependent manner prior to protease production, and via the Dectin-1 β-glucan receptor. ΔpacC mutants are defective in both contact-mediated epithelial entry and protease expression, and significantly attenuated for pathogenicity in leukopenic mice. We combined murine infection modelling, in vivo transcriptomics, and in vitro infections of human alveolar epithelia, to delineate two major, and sequentially acting, PacC-dependent processes impacting epithelial integrity in vitro and tissue invasion in the whole animal. We demonstrate that A. fumigatus spores and germlings are internalised by epithelial cells in a contact-, actin-, cell wall- and Dectin-1 dependent manner and ΔpacC mutants, which aberrantly remodel the cell wall during germinative growth, are unable to gain entry into epithelial cells, both in vitro and in vivo. We further show that PacC acts as a global transcriptional regulator of secreted molecules during growth in the leukopenic mammalian lung, and profile the full cohort of secreted gene products expressed during invasive infection. Our study reveals a combinatorial mode of tissue entry dependent upon sequential, and mechanistically distinct, perturbations of the pulmonary epithelium and demonstrates, for the first time a protective role for Dectin-1 blockade in epithelial defences. Infecting ΔpacC mutants are hypersensitive to cell wall-active antifungal agents highlighting the value of PacC signalling as a target for antifungal therapy.


Relative influence of meteorological conditions and aerosols on the lifetime of mesoscale convective systems.
Using collocated measurements from geostationary and polar-orbital satellites over tropical continents, we provide a large-scale statistical assessment of the relative influence of aerosols and meteorological conditions on the lifetime of mesoscale convective systems (MCSs). Our results show that MCSs' lifetime increases by 3-24 h when vertical wind shear (VWS) and convective available potential energy (CAPE) are moderate to high and ambient aerosol optical depth (AOD) increases by 1 SD (1σ). However, this influence is not as strong as that of CAPE, relative humidity, and VWS, which increase MCSs' lifetime by 3-30 h, 3-27 h, and 3-30 h per 1σ of these variables and explain up to 36%, 45%, and 34%, respectively, of the variance of the MCSs' lifetime. AOD explains up to 24% of the total variance of MCSs' lifetime during the decay phase. This result is physically consistent with that of the variation of the MCSs' ice water content (IWC) with aerosols, which accounts for 35% and 27% of the total variance of the IWC in convective cores and anvil, respectively, during the decay phase. The effect of aerosols on MCSs' lifetime varies between different continents. AOD appears to explain up to 20-22% of the total variance of MCSs' lifetime over equatorial South America compared with 8% over equatorial Africa. Aerosols over the Indian Ocean can explain 20% of total variance of MCSs' lifetime over South Asia because such MCSs form and develop over the ocean. These regional differences of aerosol impacts may be linked to different meteorological conditions.


Network inference with hidden units.
We derive learning rules for finding the connections between units in stochastic dynamical networks from the recorded history of a "visible'' subset of the units. We consider two models. In both of them, the visible units are binary and stochastic. In one model the "hidden'' units are continuous-valued, with sigmoidal activation functions, and in the other they are binary and stochastic like the visible ones. We derive exact learning rules for both cases. For the stochastic case, performing the exact calculation requires, in general, repeated summations over an number of configurations that grows exponentially with the size of the system and the data length, which is not feasible for large systems. We derive a mean field theory, based on a factorized ansatz for the distribution of hidden-unit states, which offers an attractive alternative for large systems. We present the results of some numerical calculations that illustrate key features of the two models and, for the stochastic case, the exact and approximate calculations.


Investigations on the oligosaccharide units of an A myeloma globulin.
The carbohydrate content of an A myeloma globulin was investigated. The carbohydrate content was found to be unchanged when the protein was isolated from the patient over a period of 18 months. The various polymeric forms of the protein contained similar proportions of carbohydrate. The A myeloma globulin contained approx. 2 residues of 6-deoxy-l-galactose (l-fucose), 14-15 of d-mannose, 12-13 of d-galactose, 12-13 of 2-acetamido-2-deoxy-d-glucose (N-acetyl-d-glucosamine), 6 of 2-acetamido-2-deoxy-d-galactose (N-acetyl-d-galactosamine) and 5 of N-acetylneuraminic acid (sialic acid), and these were distributed between six oligosaccharide units all of which were present on the heavy polypeptide chains. The oligosaccharide units showed two kinds of heterogeneity, which have been termed central and peripheral. Central heterogeneity was shown by the presence of three completely different core units, which had the following compositions: (1) 3 residues of d-galactose and 3 of 2-acetamido-2-deoxy-d-galactose, joined to protein by an O-glycosidic linkage between acetamidohexose and serine; (2) 3 residues of d-mannose, 2 of d-galactose and 3 of 2-acetamido-2-deoxy-d-glucose, joined to protein by an N-glycosidic linkage between acetamidohexose and aspartic acid; (3) 4 residues of d-mannose and 3 of 2-acetamido-2-deoxy-d-glucose with a linkage similar to that in (2). The core oligosaccharide units showed peripheral heterogeneity in the attachment of 6-deoxy-l-galactose, 2-acetamido-2-deoxy-d-glucose and N-acetylneuraminic acid. Tentative structures are proposed for these various types of oligosaccharide unit. Glycopeptides were isolated in which the sialic acid content exceeded that of d-galactose. Explanations are given for the electrophoretic mobility and staining characteristics of the various glycopeptides.


Synalpheus pinkfloydi sp. nov., a new pistol shrimp from the tropical eastern Pacific (Decapoda: Alpheidae).
A new, conspicuously coloured species of the alpheid genus Synalpheus Spence Bate, 1888, is described based on material collected on the Pacific coast of Panama. Synalpheus pinkfloydi sp. nov. is closely related to the western Atlantic S. antillensis Coutière, 1909, the two taxa being transisthmian, cryptic sister species. Both species are characterised by the distal areas of their major and minor chelae coloured in an intense, almost glowing pink-red. The morphological differences between S. pinkfloydi sp. nov. and S. antillensis Coutière, 1909 are subtle, being limited to the slightly different proportions of the merus of both chelipeds, distodorsal armature of the major cheliped merus, relative length of the antennal scaphocerite, and body size. However, they are genetically different with a 10.2% sequence divergence in COI. Based on molecular clock estimates, these transisthmian taxa diverged around 6.8-7.8 mya, i.e. well before the final closure of the Isthmus of Panama 2.5-3 mya.


ATP hydrolysis is required for cohesin's association with chromosomes.
A multi-subunit protein complex called cohesin is involved in holding sister chromatids together after DNA replication. Cohesin contains four core subunits: Smc1, Smc3, Scc1, and Scc3. Biochemical studies suggest that Smc1 and Smc3 each form 50 nm-long antiparallel coiled coils (arms) and bind to each other to form V-shaped heterodimers with globular ABC-like ATPases (created by the juxtaposition of N- and C-terminal domains) at their apices. These Smc "heads" are connected by Scc1, creating a tripartite proteinaceous ring. To investigate the role of Smc1 and Smc3's ATPase domains, we engineered smc1 and smc3 mutations predicted to abolish either ATP binding or hydrolysis. All mutations abolished Smc protein function. The binding of ATP to Smc1, but not Smc3, was essential for Scc1's association with Smc1/3 heterodimers. In contrast, mutations predicted to prevent hydrolysis of ATP bound to either head abolished cohesin's association with chromatin but not Scc1's ability to connect Smc1's head with that of Smc3. Inactivation of the Scc2/4 complex had a similar if not identical effect; namely, the production of tripartite cohesin rings that cannot associate with chromosomes. Cohesin complexes whose heads have been connected by Scc1 must hydrolyze ATP in order to associate stably with chromosomes. If chromosomal association is mediated by the topological entrapment of DNA inside cohesin's ring, then ATP hydrolysis may be responsible for creating a gate through which DNA can enter. We suggest that ATP hydrolysis drives the temporary disconnection of Scc1 from Smc heads that are needed for DNA entrapment and that this process is promoted by Scc2/4.


Type 2 diabetes-associated carotid plaque burden is increased in patients with retinopathy compared to those without retinopathy.
Cardiovascular disease (CVD) is the leading cause of mortality among subjects with type 2 diabetes (T2D), and diabetic retinopathy (DR) has been associated with an increased risk for CVD. The present study was designed to test the concept that T2D patients with DR, but without previous cardiovascular (CV) events and with normal renal function, have an increased atherosclerotic burden compared with patients without DR. A cross-sectional study was performed using patients with normal renal function (estimated glomerular filtration rate (eGFR) >60 ml/min) and without previous CV events. A total of 312 patients (men, 51%; mean age, 57 yrs; age range 40-75 yrs) were included in the study; 153 (49%) of the patients had DR. B-mode carotid ultrasound imaging was performed for all of the study subjects to measure the carotid intima-media thickness (cIMT) and carotid plaques in the common carotid artery (CCA), bifurcation and internal carotid artery (ICA). The percentage of carotid plaques in T2D patients with DR was higher than in T2D patients without DR (68% vs. 52.2%, p = 0.0045), and patients with DR had a higher prevalence of ≥2 carotid plaques (44.4% vs. 21.4%; p < 0.0001). No differences were observed in the cIMT measured at different carotid regions between the patients with or without DR. Using multivariate logistic regression (adjustment for major risk factors for atherosclerosis), DR was independently associated with mean-internal cIMT (p = 0.0176), with the presence of carotid plaques (p = 0.0366) and with carotid plaque burden (≥2 plaques; p < 0.0001). The present study shows that DR in T2D patients without CVD and with normal renal function is associated with a higher atherosclerotic burden (presence and number of plaques) in the carotid arteries. These patients may be at a higher risk for future CV events; therefore, an ultrasound examination of the carotid arteries should be considered in patients with DR for more careful and individualised CV assessment and follow-up.


Spatial-phase code-division multiple-access system with multiplexed Fourier holography switching for reconfigurable optical interconnection.
A new, to our knowledge, space-variant optical interconnection system based on a spatial-phase code-division multiple-access technique with multiplexed Fourier holography is described. In this technique a signal beam is spread over wide spatial frequencies by an M-sequence pseudorandom phase code. At a receiver side a selected signal beam is properly decoded, and at the same time its spatial pattern is shaped with a Fourier hologram, which is recorded by light that is encoded with the same M-sequence phase mask as the desired signal beam and by light whose spatial beam pattern is shaped to a signal routing pattern. Using the multiplexed holography, we can simultaneously route multisignal flows into individually specified receiver elements. The routing pattern can also be varied by means of switching the encoding phase code or replacing the hologram. We demonstrated a proof-of-principle experiment with a doubly multiplexed hologram that enables simultaneous routing of two signal beams. Using a numerical model, we showed that the proposed scheme can manage more than 250 routing patterns for one signal flow with one multiplexed hologram at a signal-to-noise ratio of ~5.


Cutting edge: GPR35/CXCR8 is the receptor of the mucosal chemokine CXCL17.
Chemokines are chemotactic cytokines that direct the traffic of leukocytes and other cells in the body. Chemokines bind to G protein-coupled receptors expressed on target cells to initiate signaling cascades and induce chemotaxis. Although the cognate receptors of most chemokines have been identified, the receptor for the mucosal chemokine CXCL17 is undefined. In this article, we show that GPR35 is the receptor of CXCL17. GPR35 is expressed in mucosal tissues, in CXCL17-responsive monocytes, and in the THP-1 monocytoid cell line. Transfection of GPR35 into Ba/F3 cells rendered them responsive to CXCL17, as measured by calcium-mobilization assays. Furthermore, GPR35 expression is downregulated in the lungs of Cxcl17(-/-) mice, which exhibit defects in macrophage recruitment to the lungs. We conclude that GPR35 is a novel chemokine receptor and suggest that it should be named CXCR8.


Synthesis and conformational properties of several maximally substituted hexa(spirotetrahydrofuranyl)cyclohexanes. Assessment Of the pronounced bias of the all-trans D(3)(d)()-symmetric isomer for total equatorial oxygen occupancy
The poly(spirotetrahydrofuranyl)cyclohexanes 1-4 were prepared in a series of steps that began with oxidative demercuration of pentaspirocyclic chloromercurials. Once the resulting alcohols were transformed into their ketones, it proved possible to cap this center with the Normant reagent and introduce the final heterocyclic ring. The cyclohexanones and the title compounds showed a strong tendency to project their C-O bonds equatorially to the maximum extent possible. The reluctance of these systems to participate in chair-to-chair conformational equilibration was made apparent during measurements to assess their coordination capability toward alkali metal ions. Although 3 was superior to its isomers, this polyether was overshadowed by 15 to an extent in excess of two powers of 10 in their relative capability to coordinate lithium cations. The synthesis, conformation, and low-level dynamic character of 5 are also detailed. To set the stereochemistry of the sixth spirotetrahydrofuranyl ring properly in this case, it was necessary to implement a novel strategy involving late-stage introduction of the oxygen atom. This protocol required intermediate formation of homoallylic alcohol 39, the epoxidation of which proceeded principally in the desired direction. X-ray crystallographic analysis of 5 established that the chair conformation which is adopted has all six C-O bonds projected equatorially. The total inability of 5 to bind to Li(+), Na(+), and K(+) denotes the existence of a substantial barrier to ring inversion. DNMR studies undertaken to assess the magnitude of this barrier demonstrated no change in high-field (1)H and (13)C line shapes up to 573K in 1, 3-([D(3)]methoxy)benzene. Consequently, 5 may qualify as the cyclohexane having the highest chair-chair conformational inversion barrier to the present time.


Pulsed indirect photoacoustic spectroscopy: application to remote detection of condensed phases.
The technique of pulsed indirect photoacoustic spectroscopy is applied to the examination of free liquid surfaces, and the prospects are assessed for remote detection and identification of chemical species in a field environment. A CO(2) laser (tunable within the 9-11-microm region) provides pulsed excitation for a variety of sample types; the resulting photoacoustic pulses are detected at ranges of the order of a few centimeters. The phenomenon is investigated as a function of parameters such as temperature, sample depth, laser-pulse energy, pulse length, and beam diameter. The results are in good agreement with a theoretical model that assumes the mechanism to be expansion of air resulting from heat conduction from the laser-heated surface of the sample under investigation. Signal and noise processing issues are discussed briefly, and the possible extension of the technique to ranges of the order of 10 m is assessed.


Androgen-glucocorticoid interactions in the era of novel prostate cancer therapy.
Great strides have been made in the treatment of castration-resistant prostate cancer (CRPC) with the development of new antiandrogens (enzalutamide) and more potent androgen synthesis inhibitors (abiraterone) that have both improved patient outcomes. These new drugs have also helped unravel the complex biology of androgen-androgen receptor driven prostate cancer and brought into prominence various mechanisms triggering the development of drug resistance and tumour cell survival despite use of androgen deprivation therapy (ADT). The complex role of glucocorticoids in the treatment, management and progression of patients with CRPC is integral to these advances. Historically, glucocorticoid treatment has resulted in both subjective and objective responses in patients with advanced-stage prostate cancer. With the use of these new therapeutic agents, however, unexpected glucocorticoid-related mechanisms that can cause iatrogenic stimulation of prostate cancer growth have emerged, which might contribute to drug resistance and disease progression despite optimal ADT. For example, the upregulation of glucocorticoid receptors (GRs) during enzalutamide therapy results in glucocorticoid-GR-mediated regulation of androgen target genes, leading to escape from enzalutamide blockade. Thus, understanding the biological role of glucocorticoids in patients with prostate cancer is of major importance in the era of new and evolving antiandrogen therapies.


Effect of Gallotannins Derived from Sedum takesimense on Tomato Bacterial Wilt.
In the process of searching antibacterial agents from plants, we discovered that the methanol extract of Sedum takesimense showed potent antibacterial activity against Ralstonia solanacearum in vitro and in vivo. Eight antibacterial gallotannins were isolated from the aerial parts of S. takesimense and identified as gallic acid, methyl gallate, 4,6-di-O-galloylarbutin, 2,6-di-O-galloylarbutin, 2,4,6-tri-O-galloyl-glucose, 1,3,4,6-tetra-O-galloyl-β-glucose, 1,2,4,6-tetra-O-galloyl-β-glucose, and 1,2,3,6-tetra-O-galloyl-β-glucose based on electrospray ionization mass spectrometry and proton nuclear magnetic resonance spectroscopy. These gallotannins displayed broad-spectrum activity against various plant-pathogenic bacteria, and the strongest in vitro antibacterial activities of these gallotannins were against R. solanacearum minimum inhibitory concentration = 0.02 to 0.10 g/liter). Among these gallotannins, methyl gallate and 1,2,3,6-tetra-O-galloyl-β-glucose showed the strongest activities. In addition, synergistic or partial synergistic effects were observed in most combinations between major antibacterial compounds. The wettable powder formulation of the S. takesimense crude extract effectively reduced the development of tomato bacterial wilt caused by R. solanacearum under greenhouse conditions for 14 days after infection. This is the first report on the isolation of antibacterial compounds from S. takesimense. These results suggest that the extract from S. takesimense or the isolated gallotannins could be used as natural bactericides for the control of tomato bacterial wilt.


Complete Genome Sequences of Four Extensively Drug-Resistant Pseudomonas aeruginosa Strains, Isolated from Adults with Ventilator-Associated Pneumonia at a Tertiary Referral Hospital in Mexico City.
Four extensively drug-resistant Pseudomonas aeruginosa strains, isolated from patients with pneumonia, were sequenced using PacBio RS-II single-molecule real-time (SMRT) technology. Genome sequence analysis identified great variability among mobile genetic elements, as well as some previously undescribed genomic islands and new variants of class 1 integrons (In1402, In1403, In1404, and In1408).


Empirical medical therapy in idiopathic male infertility: Promise or panacea?
Male factors account for 20%-50% of cases of infertility and in 25% of cases, the etiology of male infertility is unknown. Effective treatments are well-established for hypogonadotropic hypogonadism, male accessory gland infection, retrograde ejaculation, and positive antisperm antibody. However, the appropriate treatment for idiopathic male infertility is unclear. Empirical medical treatment (EMT) has been used in men with idiopathic infertility and can be divided into two categories based on the mode of action: hormonal treatment and antioxidant supplementation. Hormonal medications consist of gonadotropins, androgens, estrogen receptor blockers, and aromatase inhibitors. Antioxidants such as vitamins, zinc, and carnitines have also been widely used to reduce oxidative stress-induced spermatozoa damage. Although scientifically acceptable evidence of EMT is limited because of the lack of large, randomized, controlled studies, recent systematic reviews with meta-analyses have shown that the administration of gonadotropins, anti-estrogens, and oral antioxidants results in a significant increase in the live birth rate compared with control treatments. Therefore, all physicians who treat infertility should bear in mind that EMT can improve semen parameters and subsequent fertility potential through natural intercourse.


Non-acylated ghrelin counteracts the metabolic but not the neuroendocrine response to acylated ghrelin in humans.
Ghrelin possesses strong GH-releasing activity but also other endocrine activities including stimulation of PRL and ACTH secretion, modulation of insulin secretion and glucose metabolism. It is assumed that the GH secretagogue (GHS) receptor (GHS-R) 1a mediates ghrelin actins provided its acylation in Serine 3; in fact, acylated ghrelin only is able to exert endocrine activities. Acylated ghrelin (AG) is present in serum at a 2.5 fold lower concentration than unacylated ghrelin (UAG). UAG, however, is not biologically inactive; it shares with AG some non-endocrine actions like cardiovascular effects, modulation of cell proliferation and even some influence on adipogenesis. Thus, these actions are likely to be mediated by GHS-R subtypes able to bind ghrelin independently of its acylation. In order to further clarify whether UAG is really devoid of any endocrine action, we studied the interaction of the combined administration of AG and UAG (1.0 microg/kg i.v.) in 6 normal young volunteers (age [mean +/- SE]: 25.4 +/- 1.2 yr; BMI: 22.3 +/- 1.0 kg/m2). As expected, AG induced marked increase (p < 0.01) in circulating GH, PRL, ACTH and cortisol levels. AG administration was also followed by a decrease in insulin levels (-285.4 +/- 64.8 mU*min/l; p < 0.05) and an increase in plasma glucose levels (1068.4 +/- 390.4 mg*min/dl; p < 0.01). UAG alone did not induce any change in these parameters. UAG also failed to modify the GH, PRL, ACTH and cortisol responses to AG. However, when UAG was co-administered together with AG, no significant change in insulin (-0.5 +/- 40.9 mU*min/l) and glucose levels (455.9 +/- 88.3 mg*min/dl) was recorded anymore, indicating that the insulin and glucose response to AG has been abolished by UAG. In conclusion, non-acylated ghrelin does not affect the GH, PRL, and ACTH response to acylated ghrelin but is able to antagonize the effects of acylated ghrelin on insulin secretion and glucose levels. These findings indicate that unacylated ghrelin is metabolically active and is likely to counterbalance the influence of acylated ghrelin on insulin secretion and glucose metabolism. As GHS-R1a is not bound by unacylated ghrelin, these findings suggest that GHS receptor subtypes mediate the metabolic actions of both acylated and unacylated ghrelin.


Oxidative phosphorylation is essential for felid sperm function, but is substantially lower in cheetah (Acinonyx jubatus) compared to domestic cat (Felis catus) ejaculate.
Compared with the normospermic domestic cat, sperm metabolic function is compromised in the teratospermic cat and cheetah, but the pathway(s) involved in this deficiency are unknown. Glycolysis is essential for sperm motility, yet it appears to function normally in spermatozoa of either species regardless of structural morphology. We conducted a comparative study to further understand the mechanisms of energy production in felid spermatozoa, with the hypothesis that oxidative phosphorylation is required for normal sperm function and is impaired in teratospermic ejaculates. Electroejaculates from both species were stained with MitoTracker to quantify mitochondrial membrane potential (MMP) or were incubated to assess changes in sperm function (motility, acrosomal integrity, and lactate production) after mitochondrial inhibition with myxothiazol. Sperm midpiece dimensions also were quantified. Sperm mitochondrial fluorescence (directly proportional to MMP) was ~95% lower in the cheetah compared with the normospermic and teratospermic cat, despite the cheetah having a 10% longer midpiece. In both species, MMP was increased 5-fold in spermatozoa with retained cytoplasm compared with structurally normal cells. Inhibition of oxidative phosphorylation impaired sperm function in both species, but a 100-fold higher inhibitor concentration was required in the cat compared with the cheetah. Collectively, findings revealed that oxidative phosphorylation was required for sperm function in the domestic cat and cheetah. This pathway of energy production appeared markedly less active in the cheetah, indicating a species-specific vulnerability to mitochondrial dysfunction. The unexpected, cross-species linkage between retained cytoplasmic droplets and elevated MMP may reflect increased concentrations of metabolic enzymes or substrates in these structures.


[Science in Argentina. Where do we go from here?].
The Minister of Science described in detail his plans for the creation of closer bonds between science and society in a long term attempt to improve general quality of life. This will be accomplished by strengthening the infrastructure with diversification of the finances, multidisciplinary interrelations aiming to increase production, with special attention for social demands. One of these objectives includes the creation of research-private/public company interrelations, stressing the importance of multidisciplinary projects. Publication of results in high impact journals will always be a priority stressing the importance of basic research as a source of breakthroughs or technological inventions. The Minister also referred to the awarding of grants for scientific projects, the relation between research and production and the promotion of technological innovations. He defined three technological platforms, which are nanotechnology, biotechnology and communication. He also identified four problem/opportunity sectors, such as public health, energy sources, agro-industry and social development. Interaction between these has already led to an area of biotechnology applied to public health which will grow through translational medicine. He finally discussed the problem of patents and their importance in promoting successful business-research partnerships.


Human brain glucose metabolism may evolve during activation: findings from a modified FDG PET paradigm.
In human brain, short-term physiological stimulation results in dramatic and proportional increase in blood flow and metabolic rate of glucose but minimal change in oxygen utilization, however, with continuing stimulation, we have observed that blood flow response diminishes and oxygen utilization increases. Given the temporal limitation of conventional methods to measure glucose metabolism in the human brain, we modified [(18)F]fluorodeoxyglucose (FDG) PET paradigm to evaluate the short-term and long-term effects of visual stimulation on human brain glucose metabolism. In the present study, seven healthy volunteers each underwent three dynamic FDG PET studies: at rest and after 1 min and 15 min of visual stimulation (using reversing black-white checkerboard) which continued for only 5 min after FDG injection. We found that increase in FDG uptake in the visual cortex was attenuated by 28% when preceded by 15 min of continuous visual stimulation (p<0.001). This decline in metabolism occurred in the absence of any behavior changes in task performance. The similarity in behavior of blood flow and glucose metabolism over time supports the hypothesis that, in activated brain, blood flow is modulated by changes in cytosolic free NADH/NAD(+) ratio related to increased glycolysis. Furthermore, the observed decline in glucose metabolism may reflect a shift from glycolytic to oxidative glucose metabolism with continued activation.


Sexual differences and estrous cycle in methamphetamine-induced dopamine and serotonin depletions in the striatum of mice.
Four consecutive doses (10 mg/kg) of methamphetamine, s.c., produced a substantial striatal dopamine depletion in both sexes of BALB/c and C57BL/6J mice. Male C57BL/6J mice exhibited greater dopamine depletions in the striatum compared to female C57BL/6J mice. In contrast, male and female BALB/c mice demonstrated an equivalent magnitude of striatal dopamine depletion. Regardless of sex, C57BL/6J mice demonstrated approximately 1.4 to 2.2 times greater dopamine depletions in the striatum compared to BALB/c mice. Moreover, methamphetamine caused 4 times greater serotonin depletions in male as opposed to female BALB/c mice while sparing either sex of the C57BL/6J mice. Furthermore, female mice of both strains appeared to have the greatest basal dopamine levels during proestrus and the lowest basal dopamine levels during diestrus. Likewise, female mice of both strains exhibited the lowest dopamine depletions in the striatum when the dosing regimen of methamphetamine started at proestrus whereas the greatest dopamine depletions in the striatum occurred when the regimen started during diestrus. These results suggest that sex hormones and other modulating factors may play a role in methamphetamine-induced dopamine and serotonin neurotoxicity.


Comparison of pseudorabies virus China reference strain with emerging variants reveals independent virus evolution within specific geographic regions.
Pseudorabies virus (PRV) China reference strain Ea is genetically closely related to newly emerged variants; however, there is limited information about PRV Ea. Here, we compared PRV Ea with new variant strains by growth kinetics, genome sequencing, and protein expression analysis. Growth analysis showed that strain Ea forms smaller plaques than strain HNX. The full-length genome sequence of Ea revealed that it is clustered in the same subgroup as HNX. Ea and HNX strains exhibited similar extracellular virion protein polymorphisms, whereas strain Bartha expressed less VP26 and more GAPDH. In infected cells, strain Ea expressed high levels of IE180 protein, and Ea and HNX produced higher levels of UL21 protein than strain Bartha. These findings provide evidence that PRV China reference strain Ea is genetically closely related to the newly emerged variant strains, indicating that strain PRV China may have evolved independently leading to the emergence of a variant strain.


Improving Clinical Trial Efficiency: Thinking outside the Box.
Clinical trial design strategies have evolved over the past few years as a means to accelerate the drug development process so that the right therapies can be delivered to the right patients. Basket, umbrella, and adaptive enrichment strategies represent a class of novel designs for testing targeted therapeutics in oncology. Umbrella trials include a central infrastructure for screening and identification of patients, and focus on a single tumor type or histology with multiple subtrials, each testing a targeted therapy within a molecularly defined subset. Basket trial designs offer the possibility to include multiple molecularly defined subpopulations, often across histology or tumor types, but included in one cohesive design to evaluate the targeted therapy in question. Adaptive enrichment designs offer the potential to enrich for patients with a particular molecular feature that is predictive of benefit for the test treatment based on accumulating evidence from the trial. This review will aim to discuss the fundamentals of these design strategies, the underlying statistical framework, the logistical barriers of implementation, and, ultimately, the interpretation of the trial results. New statistical approaches, extensive multidisciplinary collaboration, and state of the art data capture technologies are needed to implement these strategies in practice. Logistical challenges to implementation arising from centralized assay testing, requirement of multiple specimens, multidisciplinary collaboration, and infrastructure requirements will also be discussed. This review will present these concepts in the context of the National Cancer Institute's precision medicine initiative trials: MATCH, ALCHEMIST, Lung MAP, as well as other trials such as FOCUS4.


Characterization of TUP1, a mediator of glucose repression in Saccharomyces cerevisiae.
The TUP1 and CYC8 (= SSN6) genes of Saccharomyces cerevisiae play a major role in glucose repression. Mutations in either TUP1 or CYC8 eliminate or reduce glucose repression of many repressible genes and induce other phenotypes, including flocculence, failure to sporulate, and sterility of MAT alpha cells. The TUP1 gene was isolated in a screen for genes that regulate mating type (V.L. MacKay, Methods Enzymol. 101:325-343, 1983). We found that a 3.5-kb restriction fragment was sufficient for complete complementation of tup1-100. The gene was further localized by insertional mutagenesis and RNA mapping. Sequence analysis of 2.9 kb of DNA including TUP1 revealed only one long open reading frame which predicts a protein of molecular weight 78,221. The predicted protein is rich in serine, threonine, and glutamine. In the carboxyl region there are six repeats of a pattern of about 43 amino acids. This same pattern of conserved residues is seen in the beta subunit of transducin and the yeast CDC4 gene product. Insertion and deletion mutants are viable, with the same range of phenotypes as for point mutants. Deletions of the 3' end of the coding region produced the same mutant phenotypes as did total deletions, suggesting that the C terminus is critical for TUP1 function. Strains with deletions in both the CYC8 and TUP1 genes are viable, with phenotypes similar to those of strains with a single deletion. A deletion mutation of TUP1 was able to suppress the snf1 mutation block on expression of the SUC2 gene encoding invertase.


Timeliness of Autism Spectrum Disorder Diagnosis and Use of Services Among U.S. Elementary School-Aged Children.
This study assessed the relationship of timeliness of autism spectrum disorder (ASD) diagnosis with current use of ASD-related services in a nationally representative sample of U.S. children. The Centers for Disease Control's (CDC's) Survey of Pathways to Diagnosis and Services was used to assess experiences of 722 children ages six to 11 with ASD. Bivariate and multivariate analyses were used to explore associations between age at ASD diagnosis and delay in ASD diagnosis and use of health services. Health services included current use of behavioral intervention (BI) therapy, school-based therapy, complementary and alternative medicine (CAM), and psychotropic medications. Mean age at ASD diagnosis was 4.4 years, and mean diagnostic delay was 2.2 years. In adjusted analysis, older age at diagnosis (≥4 versus <4) was associated with lower likelihood of current BI or school-based therapy use and higher likelihood of current psychotropic medication use. Analyses that treated age at diagnosis as a continuous variable found that likelihood of current psychotropic medication use increased with older age at diagnosis. A delay of two or more years between parents' first discussion of concerns with a provider and ASD diagnosis was associated with higher likelihood of current CAM use. Likelihood of current CAM use increased as delay in diagnosis became longer. Both older age at diagnosis and longer delay in diagnosis were associated with different health services utilization patterns among younger children with ASD. Prompt and early diagnosis may be associated with increased use of evidence-based therapies for ASD.


Kinetic Models of Cyclosporin A in Polar and Apolar Environments Reveal Multiple Congruent Conformational States.
The membrane permeability of cyclic peptides and peptidomimetics, which are generally larger and more complex than typical drug molecules, is likely strongly influenced by the conformational behavior of these compounds in polar and apolar environments. The size and complexity of peptides often limit their bioavailability, but there are known examples of peptide natural products such as cyclosporin A (CsA) that can cross cell membranes by passive diffusion. CsA is an undecapeptide with seven methylated backbone amides. Its crystal structure shows a "closed" twisted β-pleated sheet conformation with four intramolecular hydrogen bonds that is also observed in NMR measurements of CsA in chloroform. When binding to its target cyclophilin, on the other hand, CsA adopts an "open" conformation without intramolecular hydrogen bonds. In this study, we attempted to sample the complete conformational space of CsA in chloroform and in water by molecular dynamics simulations in order to better understand its conformational behavior in these two environments and to rationalize the good membrane permeability of CsA observed experimentally. From 10 μs molecular dynamics simulations in each solvent, Markov state models were constructed to characterize the metastable conformational states. The model in chloroform is compared to nuclear Overhauser effect NMR spectroscopy data reported in this study and taken from the literature. The conformational landscapes in the two solvents show significant overlap but also clearly distinct features.


Idiopathic polypoidal choroidal vasculopathy: case report.
In 1990, Yannuzzi et al. introduced a new clinical entity, "idiopathic polypoidal choroidal vasculopathy" (IPCV). There is now a more-widespread worldwide recognition of a higher prevalence and importance of this disorder. We report on the first case of IPCV in Taiwan to our knowledge. A 63-year-old Taiwanese man complained of blurred vision in his right eye for 6 months. The anterior segment and vitreous were normal except for mild nuclear cataract changes. Fundus examination of the right eye showed hard exudates and multiple reddish-orange subretinal lesions in the macula. Fundus examination of the left eye was normal. Fluorescein angiography of the right eye showed multiple hyperfluorescent granular lesions in the macula. Subretinal leakage and pooling of fluorescein were present in the temporal macula. Indocyanine green angiography disclosed multiple aneurysmal dilations at the terminals of branching vascular networks. A hyperfluorescent aneurysmal lesion corresponding to the reddish-orange nodule was observed ophthalmoscopically, and a granular lesion was observed in fluorescein angiography. Focal retinal photocoagulation was applied to the leakage area identified with fluorescein angiography. Two months after photocoagulation was performed, the subretinal fluid had gradually resolved, but a new leakage in the macula was demonstrated with fluorescein angiography. His best corrected vision in the right eye was stable.


Inaccessibility of reinforcement increases persistence and signaling behavior in the fox squirrel (Sciurus niger).
Under natural conditions, wild animals encounter situations where previously rewarded actions do not lead to reinforcement. In the laboratory, a surprising omission of reinforcement induces behavioral and emotional responses described as frustration. Frustration can lead to aggressive behaviors and to the persistence of noneffective responses, but it may also lead to new behavioral responses to a problem, a potential adaptation. We assessed the responses to inaccessible reinforcement in free-ranging fox squirrels (Sciurus niger). We trained squirrels to open a box to obtain food reinforcement, a piece of walnut. After 9 training trials, squirrels were tested in 1 of 4 conditions: a control condition with the expected reward, an alternative reinforcement (a piece of dried corn), an empty box, or a locked box. We measured the presence of signals suggesting arousal (e.g., tail flags and tail twitches) and found that squirrels performed fewer of these behaviors in the control condition and increased certain behaviors (tail flags, biting box) in the locked box condition, compared to other experimental conditions. When faced with nonreinforcement, that is, frustration, squirrels increased the number of interactions with the apparatus and spent more time interacting with the apparatus. This study of frustration responses in a free-ranging animal extends the conclusions of captive studies to the field and demonstrates that fox squirrels show short-term negatively valenced responses to the inaccessibility, omission, and change of reinforcement. (PsycINFO Database Record


Aging, Maturation and Growth of Sauropodomorph Dinosaurs as Deduced from Growth Curves Using Long Bone Histological Data: An Assessment of Methodological Constraints and Solutions.
Information on aging, maturation, and growth is important for understanding life histories of organisms. In extinct dinosaurs, such information can be derived from the histological growth record preserved in the mid-shaft cortex of long bones. Here, we construct growth models to estimate ages at death, ages at sexual maturity, ages at which individuals were fully-grown, and maximum growth rates from the growth record preserved in long bones of six sauropod dinosaur individuals (one indeterminate mamenchisaurid, two Apatosaurus sp., two indeterminate diplodocids, and one Camarasaurus sp.) and one basal sauropodomorph dinosaur individual (Plateosaurus engelhardti). Using these estimates, we establish allometries between body mass and each of these traits and compare these to extant taxa. Growth models considered for each dinosaur individual were the von Bertalanffy model, the Gompertz model, and the logistic model (LGM), all of which have inherently fixed inflection points, and the Chapman-Richards model in which the point is not fixed. We use the arithmetic mean of the age at the inflection point and of the age at which 90% of asymptotic mass is reached to assess respectively the age at sexual maturity or the age at onset of reproduction, because unambiguous indicators of maturity in Sauropodomorpha are lacking. According to an AIC-based model selection process, the LGM was the best model for our sauropodomorph sample. Allometries established are consistent with literature data on other Sauropodomorpha. All Sauropodomorpha reached full size within a time span similar to scaled-up modern mammalian megaherbivores and had similar maximum growth rates to scaled-up modern megaherbivores and ratites, but growth rates of Sauropodomorpha were lower than of an average mammal. Sauropodomorph ages at death probably were lower than that of average scaled-up ratites and megaherbivores. Sauropodomorpha were older at maturation than scaled-up ratites and average mammals, but younger than scaled-up megaherbivores.


Impact of a fitness intervention on medical students.
It is known that regular exercise improves both physical and mental health. This study sought to determine the impact of a fitness intervention on the levels of exercise and well-being of medical students. In 2011, the authors conducted a prospective cohort study involving medical students at the Johns Hopkins School of Medicine. Intervention students experienced a 7-week, team-based, fitness competition and recorded exercise data online. Incentives were given to teams reaching an average of 150 minutes per teammate per week, an exercise goal recommended by the US Department of Health and Human Services. Both groups completed baseline and follow-up surveys about physical activity and well-being, using validated scoring methods. A total of 100 students (71 in the intervention group and 29 in the control group) participated fully, recording their exercise behaviors and completing both the pre- and postsurveys. In the intervention group, the percentage of individuals successfully reaching the exercise target of 150 minutes per week varied by week from 30% to 61%. Intervention students showed a significant improvement in their International Physical Activity Questionnaire scores (1669.4 ± 154.9 vs 2013.6 ± 174.6; P = 0.02) and levels of irritability on the subsection of the Positive Affect and Negative Affect Scale score (2.2 ± 0.1 vs 2.0 ± 0.1, P = 0.03). By contrast, the control group did not show any improvements in any of these measures across the same time period (all P > 0.05). A well-orchestrated student-designed fitness intervention can effectively augment medical students' exercise practices and positively affect well-being.


Budget impact and sustainability of medical care management for persons with serious mental illnesses.
The authors assessed the 2-year outcomes, costs, and financial sustainability of a medical care management intervention for community mental health settings. A total of 407 psychiatric outpatients with serious mental illnesses were randomly assigned to usual care or to a medical care manager who provided care coordination and education. Two-year follow-up chart reviews and interviews assessed quality and outcomes of care, as well as costs from both the health system and managerial perspectives. Sustained improvements were observed in the intervention group in quality of primary care preventive services, quality of cardiometabolic care, and mental health-related quality of life. From a health system perspective, by year 2, the mean per-patient total costs for the intervention group were $932 (95% CI=-1,973 to 102) less than for the usual care group, with a 92.3% probability that the program was associated with lower costs than usual care. From the community mental health center perspective, the program would break even (i.e., revenues would cover setup costs) if 58% or more of clients had Medicaid or another form of insurance. Given that only 40.5% of clients in this study had Medicaid, the program was not sustainable after grant funding ended. The positive long-term outcomes and favorable cost profile provide evidence of the potential value of this model. However, the discrepancy between health system and managerial cost perspectives limited the program's financial sustainability. With anticipated insurance expansions under health reform, there is likely to be a stronger business case for safety net organizations considering implementing evidence-based interventions such as the one examined in this study.


Lower-extremity sclerotomes: clinical applications and the modified mayo block.
A sclerotome is an anatomical concept that defines an area of bone supplied by a single spinal nerve. Similar to the familiar dermatomes, sclerotomes provide an element of depth to the sensory innervation of the lower extremity based on the deep fascia as an embryologic boundary. Anatomical knowledge of sclerotomes can be used clinically in the diagnosis and treatment of pain and in the perioperative setting. Specifically, a modified version of the classic Mayo block is presented to highlight an active anatomical approach to peripheral nerve blockade.


Neutrophil Resolvin E1 Receptor Expression and Function in Type 2 Diabetes.
Unresolved inflammation is key in linking metabolic dysregulation and the immune system in type 2 diabetes. Successful regulation of acute inflammation requires biosynthesis of specialized proresolving lipid mediators, such as E-series resolvin (RvE) 1, and activation of cognate G protein-coupled receptors. RvE1 binds to leukotriene B4 (BLT-1) on neutrophils and to ERV-1/ChemR23 on monocyte/macrophages. We show novel actions of RvE1 and expression patterns of neutrophil receptors in type 2 diabetes. Neutrophils from healthy subjects express functional BLT-1, low levels of minimally functional ERV-1, and inversed coexpression when compared to neutrophils from type 2 diabetes subjects. Stimulation with TNF-α or LPS increased the expression of ERV-1 by healthy and diabetic neutrophils. RvE1 counteracted LPS and TNF-α induction of ERV-1 overexpression and endogenous diabetic overexpression, activating phagocytosis and resolution signals. Functional ERV-1 was determined by phosphorylation of the signaling protein ribosomal S6. Receptor-antagonism experiments revealed that the increase in phosphorylation of ribosomal S6 was mediated by BLT-1 in healthy subject neutrophils and by ERV-1 in diabetes. Metabololipidomics reveal a proinflammatory profile in diabetic serum. Cell phagocytosis is impaired in type 2 diabetes and requires RvE1 for activation. The dose of RvE1 required to activate resolution signals in type 2 diabetic neutrophils was significantly higher than in healthy controls. RvE1 rescues the dysregulation seen on neutrophil receptor profile and, following a therapeutic dosage, activates phagocytosis and resolution signals in type 2 diabetes. These findings reveal the importance of resolution receptors in health, disease, and dysregulation of inflammation in type 2 diabetes.


Upregulated sirtuin 1 by miRNA-34a is required for smooth muscle cell differentiation from pluripotent stem cells.
microRNA-34a (miR-34a) and sirtuin 1 (SirT1) have been extensively studied in tumour biology and longevity/aging, but little is known about their functional roles in smooth muscle cell (SMC) differentiation from pluripotent stem cells. Using well-established SMC differentiation models, we have demonstrated that miR-34a has an important role in SMC differentiation from murine and human embryonic stem cells. Surprisingly, deacetylase sirtuin 1 (SirT1), one of the top predicted targets, was positively regulated by miR-34a during SMC differentiation. Mechanistically, we demonstrated that miR-34a promoted differentiating stem cells' arrest at G0/G1 phase and observed a significantly decreased incorporation of miR-34a and SirT1 RNA into Ago2-RISC complex upon SMC differentiation. Importantly, we have identified SirT1 as a transcriptional activator in the regulation of SMC gene programme. Finally, our data showed that SirT1 modulated the enrichment of H3K9 tri-methylation around the SMC gene-promoter regions. Taken together, our data reveal a specific regulatory pathway that miR-34a positively regulates its target gene SirT1 in a cellular context-dependent and sequence-specific manner and suggest a functional role for this pathway in SMC differentiation from stem cells in vitro and in vivo.


Far field diffraction patterns of circular gratings.
The fine structure of the annular images of circular gratings is analyzed in terms of diffraction patterns of axicon pairs. Each image arises substantially from only one converging and one diverging axicon of equal deflection angle. Single main lobe, symmetric double main lobe, and various intermediate asymmetric double lobe structures are obtained depending on the phase relationship and strengths of the two axicon beams, which in turn depend on the design of the circular grating. Approximate expressions are derived for the intensity distribution and energy content of the symmetric single and double lobe images.


The cell adhesion protein P-selectin glycoprotein ligand-1 is a substrate for the aspartyl protease BACE1.
The aspartyl protease BACE1 cleaves the amyloid precursor protein and the sialyltransferase ST6Gal I and is important in the pathogenesis of Alzheimer's disease. The normal function of BACE1 and additional physiological substrates have not been identified. Here we show that BACE1 acts on the P-selectin glycoprotein ligand 1 (PSGL-1), which mediates leukocyte adhesion in inflammatory reactions. In human monocytic U937 and human embryonic kidney 293 cells expressing endogenous or transfected BACE1, PSGL-1 was cleaved by BACE1 to generate a soluble ectodomain and a C-terminal transmembrane fragment. No evidence of the cleavage fragment was seen in primary cells derived from mice deficient in BACE1. By using deletion constructs and enzymatic deglycosylation of the C-terminal PSGL-1 fragments, the cleavage site in PSGL-1 was mapped to the juxtamembrane region within the ectodomain. In an in vitro assay BACE1 catalyzed the formation of the PSGL-1 products seen in vivo. The cleavage occurred at a Leu-Ser peptide bond as identified by mass spectrometry using a synthetic peptide. We conclude that PSGL-1 is an additional substrate for BACE1.


Deciphering complex metabolite mixtures by unsupervised and supervised substructure discovery and semi-automated annotation from MS/MS spectra.
Complex metabolite mixtures are challenging to unravel. Mass spectrometry (MS) is a widely used and sensitive technique for obtaining structural information of complex mixtures. However, just knowing the molecular masses of the mixture's constituents is almost always insufficient for confident assignment of the associated chemical structures. Structural information can be augmented through MS fragmentation experiments whereby detected metabolites are fragmented, giving rise to MS/MS spectra. However, how can we maximize the structural information we gain from fragmentation spectra? We recently proposed a substructure-based strategy to enhance metabolite annotation for complex mixtures by considering metabolites as the sum of (bio)chemically relevant moieties that we can detect through mass spectrometry fragmentation approaches. Our MS2LDA tool allows us to discover - unsupervised - groups of mass fragments and/or neutral losses, termed Mass2Motifs, that often correspond to substructures. After manual annotation, these Mass2Motifs can be used in subsequent MS2LDA analyses of new datasets, thereby providing structural annotations for many molecules that are not present in spectral databases. Here, we describe how additional strategies, taking advantage of (i) combinatorial in silico matching of experimental mass features to substructures of candidate molecules, and (ii) automated machine learning classification of molecules, can facilitate semi-automated annotation of substructures. We show how our approach accelerates the Mass2Motif annotation process and therefore broadens the chemical space spanned by characterized motifs. Our machine learning model used to classify fragmentation spectra learns the relationships between fragment spectra and chemical features. Classification prediction on these features can be aggregated for all molecules that contribute to a particular Mass2Motif and guide Mass2Motif annotations. To make annotated Mass2Motifs available to the community, we also present MotifDB: an open database of Mass2Motifs that can be browsed and accessed programmatically through an Application Programming Interface (API). MotifDB is integrated within ms2lda.org, allowing users to efficiently search for characterized motifs in their own experiments. We expect that with an increasing number of Mass2Motif annotations available through a growing database, we can more quickly gain insight into the constituents of complex mixtures. This will allow prioritization towards novel or unexpected chemistries and faster recognition of known biochemical building blocks.


A dedicated scholarly research program in an adult and pediatric neurology residency program.
To describe and assess the effectiveness of a formal scholarly activity program for a highly integrated adult and pediatric neurology residency program. Starting in 2011, all graduating residents were required to complete at least one form of scholarly activity broadly defined to include peer-reviewed publications or presentations at scientific meetings of formally mentored projects. The scholarly activity program was administered by the associate residency training director and included an expanded journal club, guided mentorship, a required grand rounds platform presentation, and annual awards for the most scholarly and seminal research findings. We compared scholarly output and mentorship for residents graduating within a 5-year period following program initiation (2011-2015) and during the preceding 5-year preprogram baseline period (2005-2009). Participation in scholarship increased from the preprogram baseline (24 of 53 graduating residents, 45.3%) to the postprogram period (47 of 57 graduating residents, 82.1%, p < 0.0001). Total scholarly output more than doubled from 49 activities preprogram (0.92/resident) to 139 postprogram (2.44/resident, p = 0.0002). The proportions of resident participation increased for case reports (20.8% vs 66.7%, p < 0.0001) and clinical research (17.0% vs 38.6%, p = 0.012), but were similar for laboratory research and topical reviews. The mean activities per resident increased for published abstracts (0.15 ± 0.41 to 1.26 ± 1.41, p < 0.0001), manuscripts (0.75 ± 1.37 to 1.00 ± 1.40, p = 0.36), and book chapters (0.02 ± 0.14 to 0.18 ± 0.60, p = 0.07). Rates of resident participation as first authors increased from 30.2% to 71.9% (p < 0.0001). The number of individual faculty mentors increased from 36 (preprogram) to 44 (postprogram). Our multifaceted program, designed to enhance resident and faculty engagement in scholarship, was associated with increased academic output and an expanded mentorship pool. The program was particularly effective at encouraging presentations at scientific meetings. Longitudinal analysis will determine whether such a program portfolio inspires an increase in academic careers involving neuroscience-oriented research.


Microbial community shifts in the oxic-settling-anoxic process in response to changes to sludge interchange ratio.
This particular study set out to demonstrate alterations on the microbial community of the oxic-settling-anaerobic/anoxic (OSA) process treating real domestic wastewater by changing interchange ratios (IRs). The sludge yield of systems operated at different IRs (1/13, 1/17 and 1/20) to assess sludge reduction was used to analyze microbial community composition variations. The highest IR (1/13) resulted in the highest sludge reduction (52.1%), while the OSA systems with IR of 1/17 and 1/20 reduced sludge production by 37.4% and 35.5%, respectively, in comparison to conventional systems. 16S rRNA gene amplicon sequencing analysis showed that the bacterial communities were composed of similar phylogenetic groups, Proteobacteria, Acidobacteria, and Bacteroidetes being dominant. The relative abundances differed due to the applied IRs. The highest abundance of Actinobacteria was determined at the highest IR (1/13) and increasing of the HRT to 1/20 caused a significant reduction in Actinobacteria species and the lowest abundance (6%) was determined in the OSA systems. The abundant of Thiothrix species that are boosted in the OSA trials may have a vital role in OSA systems, where its abundance was below the detection limits in the seed sludge sample. Therefore, they could be used as bioindicators in the OSA system.


Metabolic effects of cachectin/tumor necrosis factor are modified by site of production. Cachectin/tumor necrosis factor-secreting tumor in skeletal muscle induces chronic cachexia, while implantation in brain induces predominantly acute anorexia.
We have developed a murine model of wasting by injecting intracerebrally cells which continuously secrete h-cachectin/TNF (CHO-TNF) to: (a) determine the effects of cachectin/TNF produced continuously in the central nervous system (CNS), and (b) compare the metabolic effects of cachectin/TNF-secreting tumor in the brain to the cachexia caused by CHO-TNF tumor in peripheral tissue (IM). Intracerebral CHO-TNF tumors produced increased serum h-cachectin/TNF levels with lethal hypophagia and weight loss (mean survival time of 11 d); these changes were not observed in association with nonsecretory control brain tumors. The metabolic consequences of intracerebral cachectin/TNF production were indistinguishable from acute, lethal starvation: whole-body lipid content was decreased significantly but protein was conserved. Although intramuscular cachectin/TNF-secreting tumors caused similar increases of serum h-cachectin/TNF levels, profound anorexia did not develop; wasting developed after a longer period of tumor burden (50 d) with classical signs of cachexia (i.e., anemia and depletion of both protein and lipid). These studies provide a reproducible animal model of site-specific cytokine production and suggest that, regardless of serum levels, cachectin/TNF produced locally in brain influences both the rate of development of wasting and its net metabolic effects.


Prevalence of Voice Disorders in Singers: Systematic Review and Meta-Analysis.
The study aimed to review the prevalence of self-reported voice disorders in singers. The study is a systematic review and meta-analysis. A systematic review of five major scientific databases was conducted. An extensive search strategy was used considering the rules of each database. Original articles were included only if they had data related to self-perception of dysphonia in the past. Furthermore, heterogeneity and its relative significance were assessed. There were 2371 articles identified; duplicates were deleted, screenings were conducted, and inclusion and exclusion criteria were applied. The final analysis was conducted on 11 studies. The most implemented instruments for the study were customized questionnaires. The findings about singing styles, voice use, and age were found to be different among subjects. The overall prevalence of self-reported dysphonia in singers was 46.09% (95% confidence interval: 38.16-54.12). The heterogeneity was considerable among the studied samples (I2 = 90.59%). Four groups were then established-students, teachers, classical, and nonclassical-and compared regarding overall prevalence (21.76% in students, and significantly higher and nondifferent in the other three groups, 55.15%, 40.53%, and 46.96%, respectively) and heterogeneity (low only for the students' studies). Although with low homogeneity, singers present a high prevalence of self-perceived dysphonia over their careers. Singing students were the group with a lower prevalence. On the other hand, traditional and popular music singers, as well as singing teachers, revealed significantly higher prevalence of self-perceived dysphonia. Overall, singers are likely to report voice disorders, no matter their singing style or skills. This highlights the need of a preventive approach to address voice disorders in traditional and untrained singers.


AA amyloidosis in the renal allograft: a report of two cases and review of the literature.
AA amyloidosis is a disorder characterized by the abnormal formation, accumulation and systemic deposition of fibrillary material that frequently involves the kidney. Recurrent AA amyloidosis in the renal allograft has been documented in patients with tuberculosis, familial Mediterranean fever, ankylosing spondylitis, chronic pyelonephritis and rheumatoid arthritis. De novo AA amyloidosis is rarely described. We report two cases of AA amyloidosis in the renal allograft. Our first case is a 47-year-old male with a history of ankylosing spondylitis who developed end-stage renal disease reportedly from tubulointerstitial nephritis from non-steroidal anti-inflammatory agent use. A biopsy was never performed. One year after transplantation, AA amyloidosis was identified in the femoral head and 8 years post-transplantation, AA amyloidosis was identified in the renal allograft. He was treated with colchicine and adalimumab and has stable renal function at 1 year-follow-up. Our second case is a 57-year-old male with a long history of intravenous drug use and hepatitis C infection who developed end-stage kidney disease due to AA amyloidosis. Our second patient's course was complicated by renal adenovirus, pulmonary aspergillosis and hepatitis C with AA amyloidosis subsequently being identified in the allograft 2.5 years post-transplantation. Renal allograft function remains stable 4-years post-transplantation. These reports describe clinical and pathologic features of two cases of AA amyloidosis presenting with proteinuria and focal involvement of the renal allograft.


Stacking sequences of black phosphorous allotropes and the corresponding few-layer phosphorenes.
Possible bulk black phosphorus (BP) allotropes are constructed based on single-layer BP with various stacking sequences. Our stacking algorithm shows that there are eight possible allotropes with two stacking layers in their unit cells possessing relatively high symmetries, and six of them are retained after structural relaxation using a van der Waals correction of optB88-vdW. The AF, AG, and AH bulk structures are presented for the first time. The structural relationship of these configurations has been explained via an interlayer slipping process. The total energy of the AF allotrope is closest to the most stable bulk BP structure (AB stacking) among all explored 2-layer stacked bulk structures. The calculated band structure of the AF allotrope using HSE06 shows a direct band gap of 0.48 eV with anisotropic electronic structures. We also presented six possible BP allotropes with three stacking layers in their unit cells. The newly reported AAF and ABC stacked structures show semiconducting and metallic features, respectively. After the bulk structures were explored, we further built the corresponding few-layer phosphorene structures and investigated their electronic properties. The results show that all the few-layer phosphorenes show semiconducting features. The AE, AAE, and AEA phosphorenes have indirect band gaps while the other explored phosphorenes possess direct band gaps located at the Γ point.


In vitro relationship between magnesium and insulin secretion.
Magnesium, the most abundant intracellular divalent cation, is known to play an essential role in many normal cell functions, such as fluid and electrolyte transport, enzyme activity, and cell proliferation. It is known that magnesium deficiency or hypomagnesaemia induces hyperinsulinism, while hypermagnesaemia inhibits insulin secretion: however, the mechanism controlling the intracellular level of free magnesium and its role in the insulin-secretory mechanism of pancreatic beta cells (RIN m5F cells) are still unclear. Using a fluorescent indicator (mag-fura-2) we have found that the influx of magnesium appears to be voltage-dependent and is sensitive to blockade of the voltage-dependent calcium channel in RIN m5F cells; the efflux of magnesium appears to be voltage- and cyclic AMP-independent. We have observed that depletion of extracellular magnesium potentiates insulin secretion from RIN m5F cells. This finding has led us to speculate on two possible mechanisms through which extracellular magnesium participates in the regulation of insulin secretion: (1) extracellular magnesium may regulate ATP-sensitive potassium channels, and (2) extracellular magnesium may act as a competitive inhibitor of the calcium influx mediated through the voltage-dependent calcium channel. In addition to the role of extracellular magnesium, the changes in the intracellular levels of free magnesium observed during secretagogue-stimulated insulin secretion may alter magnesium-sensitive enzymes or systems, which may play regulatory roles in signal transduction. Elucidation of the role of magnesium in the insulin secretory mechanism will be beneficial for understanding the insulin secretory mechanism of pancreatic beta cells and may be helpful in treating insulin-related abnormalities in patients with hypo- or hypermagnesaemia.