Self-probed ptychography from semiconductor high-harmonic generation.
We demonstrate a method to image an object using a self-probing approach based on semiconductor high-harmonic generation. On the one hand, ptychography enables high-resolution imaging from the coherent light diffracted by an object. On the other hand, high-harmonic generation from crystals is emerging as a new source of extreme-ultraviolet ultrafast coherent light. We combine these two techniques by performing ptychography measurements with nanopatterned crystals serving as the object as well as the generation medium of the harmonics. We demonstrate that this strong field in situ approach can provide structural information about an object. With the future developments of crystal high harmonics as a compact short-wavelength light source, our demonstration can be an innovative approach for nanoscale imaging of photonic and electronic devices in research and industry.


Lidocaine enhances Galphai protein function.
Local anesthetics inhibit several G protein-coupled receptors by interaction with the Galphaq protein subunit. It is not known whether this effect on G protein function can be extrapolated to other classes of G proteins. The authors investigated interactions of lidocaine with the human adenosine 1 receptor (hA1R)-coupled signaling pathway. Activated A1Rs couple to adenylate cyclase via the pertussis toxin sensitive Galphai protein, thereby decreasing cyclic adenosine monophosphate formation. A1Rs are widely expressed and abundant in the spinal cord, brain, and heart. Interactions of LAs with the hA1R-coupled transduction cascade therefore might produce a broad range of clinically relevant effects. The function of hA1Rs stably expressed in Chinese hamster ovary cells was determined with assays of cyclic adenosine monophosphate, receptor binding, and guanosine diphosphate/guanosine triphosphate gamma35S exchange by using reconstituted defined G protein subunits. Involvement of phosphodiesterase and Galphai was characterized by using the phosphodiesterase inhibitor rolipram and pertussis toxin, respectively. Lidocaine (10-9-10-1 M) had no significant effects on agonist or antagonist binding to the hA1R or on receptor-G protein interactions. However, cyclic adenosine monophosphate levels were reduced significantly to 50% by the LAs, even in the absence of an A1R agonist or presence of an A1R antagonist. This effect was unaffected by rolipram (10 mum), but abolished completely by pretreatment with pertussis toxin, which inactivates the Galphai protein. Therefore, the main target site for LAs in this pathway is located upstream from adenylate cyclase. Lidocaine potentiates Galphai-coupled A1R signaling by reducing cyclic adenosine monophosphate production. The study suggests an interaction site for LAs in a Galphai-coupled signaling pathway, with the Galphai protein representing the prime candidate. Taken together with previous results showing inhibitory LA interactions on the Galphaq protein subunit, the data in the current study support the hypothesis that specific G protein subunits represent alternative sites of LA action.


An early hand-wrist atlas and its implications for secular change in bone age.
A little-known 'Skiagraphic Atlas' documenting radiographic changes in the bones of the hand and wrist was published by Poland in 1898. Nineteen skiagrams of British children, 1-17 years of age, were included. Tanner-Whitehouse-2 bone ages were assessed for 15 individuals in this series and differences from chronological ages were calculated. Mean differences for 20-bone, RUS and carpal bone ages are significantly larger than zero. Hence, these children were relatively advanced compared to present children. Comparisons of Poland's expected ages for onset and fusion of secondary ossification centres in the hand and wrist with more recent reference data indicated a secular increase in the rates of skeletal maturation of approximately 0.22-0.66 year/decade, with relatively greater change in expected ages of fusion.


Machine Learning Models Predict the Need of Amputation and/or Peripheral Artery Revascularization in Hypertensive Patients Within 7-Years Follow-Up.
Lower extremity amputation and requirement of peripheral artery revascularization are common outcomes of undiagnosed peripheral artery disease patients. In the current work, prediction models for the need of amputation or peripheral revascularization focused on hypertensive patients within seven years follow up are employed. We applied machine learning (ML) models using classifiers such as Extreme Gradient Boost (XGBoost), Random Forest (RF) and Adaptive Boost (AdaBoost), that will allow clinicians to identify the patients at risk of these two endpoints using simple clinical data. We used the non-interventional cohort of the getABI study in the primary care setting, selecting 4,191 hypertensive patients out of 6,474 patients with age over 65 years old and followed up for vascular events or death up to 7 years. During this follow up period, 150 patients underwent either amputation or peripheral revascularization or both. Accuracy, Specificity, Sensitivity and Area under the receiver operating characteristic curve (AUC) were estimated for each machine learning model. The results demonstrate Random Forest as the most accurate model for the prediction of the composite endpoint in hypertensive patients within 7 years follow-up, achieving 73.27 % accuracy.Clinical Relevance-This study assists clinicians to better predict and treat these serious outcomes, amputation and peripheral revascularization in hypertensive patients.


The Office Work and Stretch Training (OST) Study: An Individualized and Standardized Approach to Improve the Quality of Life in Office Workers.
In the context of workplace health promotion, physical activity programs have been shown to reduce musculoskeletal diseases and stress, and to improve the quality of life. The aim of this study was to examine the effects of using the "five-Business" stretch training device for office workers on their quality of life. A total of 313 office workers (173m/137f) participated voluntarily in this intervention-control study with an average age of 43.37 ± 11.24 (SD) years, 175.37 ± 9.35 cm in height and 75.76 ± 15.23 kg in weight, with an average BMI of 24.5 ± 3.81 kg/m2. The participants completed the stretch training twice a week for approximately 10 minutes for a duration of 12 weeks. The SF-36 questionnaire was used to evaluate the effectiveness of the intervention at baseline and after 12 weeks. Significantly improved outcomes in mental sum score (p = 0.008), physical functioning (p < 0.001), bodily pain (p = 0.01), vitality (p = 0.025), role limitations due to physical problems (p = 0.018) and mental health (p = 0.012) were shown after the stretching training. The results suggest that a 12-week stretching program for office desk workers is suitable to improve significantly their health-related quality of life.


The Role of Social Support in Concussion Disclosure in United States Air Force Academy Cadets: A Mixed Methods Approach.
United States Air Force Academy (USAFA) cadets are at risk for sustaining concussions; however, several factors inhibit disclosure. We aimed to better understand the role of social support in concussion disclosure. We used a mixed methods approach with an electronic survey and interviews. The survey used a 9-point Likert scale (1 = strongly disagree and 9 = strongly agree) to assess concussion disclosure. Survey items asked cadets whom they felt most comfortable disclosing a concussion or bell-ringer/ding to, how quickly they would seek medical attention for a suspected concussion or bell-ringer/ding, whether cadets would be generally supportive/unsupportive of another cadet disclosing a concussion to medical staff, and whether other cadets important to them would be generally supportive/unsupportive if they reported a concussion to medical staff. Two multivariate linear regressions, one for concussion and one for bell-ringers/dings, were calculated to determine whether cadet choice of the person they felt most comfortable disclosing a concussion or bell-ringer/ding to predicted whether they would immediately seek medical attention for either condition. Choice of person included Air Officer Commanding (AOC)/Academy Military Trainer (AMT), upper classmen, cadet who had recovered from a concussion, cadet, closest friend, teammate, and squadmate. Descriptive analyses assessed whether cadets felt supported or unsupported by other cadets and by those who were important to them concerning concussion reporting. Thirty-four semi-structured interviews were conducted with cadets to explore their views on concussion disclosure. Increased comfort with disclosing a suspected concussion to an AOC/AMT had higher agreement seeking medical attention (β = 0.28, P < .001). For every 1-point increase in being comfortable disclosing a potential bell-ringer/ding to an AOC/AMT (β = 0.272, P < .001), squadmate (β = 0.241, P = .002), and teammate (β = -0.182, P = .035), agreement for seeking medical attention immediately increased 0.27, increased 0.24, and decreased 0.18, respectively. Interviews indicated cadets would often speak to a peer before seeking medical attention. Our results suggested that cadets felt comfortable reporting a suspected concussion or bell-ringer/ding to various peers and that those peers would be supportive of that choice, indicating social support. Future interventions should include educating cadets that peers may come to them, especially if they are AOCs/AMTs or squadmates.


Long-term treatment effects in chronic myeloid leukemia.
We propose and analyze a simplified version of a partial differential equation (PDE) model for chronic myeloid leukemia (CML) derived from an agent-based model proposed by Roeder et al. This model describes the proliferation and differentiation of leukemic stem cells in the bone marrow and the effect of the drug Imatinib on these cells. We first simplify the PDE model by noting that most of the dynamics occurs in a subspace of the original 2D state space. Then we determine the dominant eigenvalue of the corresponding linearized system that controls the long-term behavior of solutions. We mathematically show a non-monotonous dependence of the dominant eigenvalue with respect to treatment dose, with the existence of a unique minimal negative eigenvalue. In terms of CML treatment, this shows that there is a unique dose that maximizes the decay rate of the CML tumor load over long time scales. Moreover this unique dose is lower than the dose that maximizes the initial tumor load decay. Numerical simulations of the full model confirm that this phenomenon is not an artifact of the simplification. Therefore, while optimal asymptotic dosage might not be the best one at short time scales, our results raise interesting perspectives in terms of strategies for achieving and improving long-term deep response.


Aeromicrobium lacus sp. nov., a novel actinobacterium isolated from a drinking-water reservoir.
An actinobacterial strain, designated CPCC 204604T, was isolated from a water sample collected from Sancha Lake, a drinking-water reservoir in Sichuan Province, south-west China. Cells were strictly aerobic, Gram-stain-positive, non-spore-forming and non-motile. Polyphasic taxonomic analysis indicated that galactose and ribose were detected as the diagnostic sugars in the whole-cell hydrolysates, and ll-diaminopimelic acid was the diagnostic diamino acid in cell-wall peptidoglycan. MK-9(H4) was the predominant menaquinone. The phospholipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and an unidentified phospholipid. The major fatty acids were C16 : 0, C16 : 0 2-OH, C18 : 1ω9c, C18 : 0 and 10-methyl C18:0. The genomic DNA G+C content was 71.2 mol%. In the phylogenetic tree based on 16S rRNA gene sequences, strain CPCC 204604T formed a distinct clade with Aeromicrobium ponti DSM 19178T within the lineage of the genus Aeromicrobium, with the highest 16S rRNA gene sequence similarity of 98.3 % to its closest neighbour. On the basis of the genotypic and phenotypic data, we conclude that strain CPCC 204604T represents a novel species of the genus Aeromicrobium, for which the name Aeromicrobiumlacus sp. nov. is proposed with strain CPCC 204604T (=NBRC 112936T=DSM 105424T) as the type strain.


Bioelectric signaling regulates size in zebrafish fins.
The scaling relationship between the size of an appendage or organ and that of the body as a whole is tightly regulated during animal development. If a structure grows at a different rate than the rest of the body, this process is termed allometric growth. The zebrafish another longfin (alf) mutant shows allometric growth resulting in proportionally enlarged fins and barbels. We took advantage of this mutant to study the regulation of size in vertebrates. Here, we show that alf mutants carry gain-of-function mutations in kcnk5b, a gene encoding a two-pore domain potassium (K(+)) channel. Electrophysiological analysis in Xenopus oocytes reveals that these mutations cause an increase in K(+) conductance of the channel and lead to hyperpolarization of the cell. Further, somatic transgenesis experiments indicate that kcnk5b acts locally within the mesenchyme of fins and barbels to specify appendage size. Finally, we show that the channel requires the ability to conduct K(+) ions to increase the size of these structures. Our results provide evidence for a role of bioelectric signaling through K(+) channels in the regulation of allometric scaling and coordination of growth in the zebrafish.


[Surgical treatment of external pancreatic fistulas].
An analysis was performed of surgical treatment of 27 patients presenting with fibrodegenerative form of chronic pancreatitis complicated by the presence of an external pancreatic fistula. A diagnostic algoryhthm has been developed permitting the determination of necessity to use surgical intervention prior to conservative treatment. Tactical approaches are set down to be used in a surgical treatment of the above patients.


Forgetting the once-seen face: estimating the strength of an eyewitness's memory representation.
The fidelity of an eyewitness's memory representation is an issue of paramount forensic concern. Psychological science has been unable to offer more than vague generalities concerning the relation of retention interval to memory trace strength for the once-seen face. A meta-analysis of 53 facial memory studies produced a highly reliable association (r=.18, d=0.37) between longer retention intervals and positive forgetting of once-seen faces, an effect equally strong for both face recognition and eyewitness identification studies. W. A. Wickelgren's (1974, 1975, 1977) theory of recognition memory provided statistically satisfactory fits to 11 different empirical forgetting functions. Applied to the results of field studies of eyewitness memory, the theory yields predictions relevant to fact finders' evaluations of eyewitness credibility. A plausible upper limit for witness initial memory strength corresponds to a probability of .67 of being correct on a fair six-person lineup. Furthermore, not only can the percentage of remaining memory strength be determined for any retention interval, but this strength estimate can be translated into an estimated probability of being correct on a fair lineup of a specified size.


Proopiomelanocortin-related peptides and methionine enkephalin in human follicular fluid: changes during the menstrual cycle.
Several studies indicate the presence of different pituitary hormones or neuropeptides in ovarian follicular fluid from various species. Recently our group showed that the ovarian follicular fluid of health women contains two of the endogenous opioid peptides, beta-endorphin and methionine enkephalin, in concentrations that are tenfold to twentyfold higher than in circulating plasma. The presence of immunoreactive beta-lipotropin was also shown. The aim of the present study was to evaluate whether adrenocorticotropic hormone, which in pituitary cells is synthesized from proopiomelanocortin such as beta-endorphin and beta-lipotropin, is also present in follicular fluid and the possible changes of proopiomelanocortin-related peptides during the menstrual cycle. Concentrations of beta-endorphin, methionine enkephalin, adrenocorticotropic hormone, and beta-lipotropin were measured in 60 healthy menstruating women at different periods of the menstrual cycle (20 during the follicular, 22 in the preovulatory days, and 18 during the luteal phase). Thirteen women participated in an in vitro fertilization program and thus received clomiphene citrate (100 mg/day from the fifth to the ninth day) plus 5000 IU human chorionic gonadotropin before starting the program. All samples were collected at laparoscopy under general anesthesia. In another eight patients fluid was collected from follicular cysts. Peptides were extracted on octadodecasilyl silica columns with 80% methanol in 0.5 mol/L acetic acid. The identity of follicular fluid beta-endorphin, methionine enkephalin, adrenocorticotropic hormone, and beta-lipotropin and standard peptides was demonstrated with high-pressure liquid chromatography. Peptide concentrations were measured in extracts by radioimmunoassays either directly by (methionine enkephalin and adrenocorticotropic hormone) or after (beta-endorphin and beta-lipotropin) gel filtration on Sephadex G-75. The concentrations of methionine enkephalin, adrenocorticotropic hormone, and beta-lipotropin were similar in the different periods of the cycle. Conversely, beta-endorphin concentrations were significantly higher in preovulatory days than in the other periods; no differences were evident between spontaneous and stimulated cycles. These results indicate that proopiomelanocortin-related peptides are present in the follicular fluid and that beta-endorphin concentrations change during the menstrual cycle, with the highest values occurring in the preovulatory follicle.


Clinical Assessment of the DiaSorin LIAISON SARS-CoV-2 Ag Chemiluminescence Immunoassay.
Due to the large volume of tests needed in a relatively short time for screening and diagnosing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, antigen immunoassays may provide a potential supplement to molecular testing. This study was aimed to assess the clinical preference of DiaSorin LIAISON SARS-CoV-2 Ag chemiluminescence immunoassay. An upper respiratory specimen was collected in a series of patients referred to the Laboratory Medicine service of Pederzoli Hospital (Peschiera del Garda, Verona, Italy) for screening or diagnosis of SARS-CoV-2 infection. Nasopharyngeal samples were assayed with DiaSorin LIAISON SARS-CoV-2 Ag test and Altona Diagnostics RealStar® SARS-CoV-2 RT-PCR Kit. The final study population consisted of 421 patients (median age, 48 years; 227 women), 301 (71.5%) with positive result of molecular testing, and 126 (29.9%) with cycle threshold (Ct) values of both E and S genes <29.5, thus reflecting higher infectivity. The area under the curve of DiaSorin LIAISON SARS-CoV-2 Ag test 0.82 (95% CI, 0.79-0.86) for sample positivity and 0.98 for higher sample infectivity (95% CI, 0.97 to 0.99). The optimal cut-off for sample positivity was 82 TCID50/mL (0.78 sensitivity, 0.73 specificity and 77% diagnostic accuracy), whilst that for identifying samples associated with a high infective risk was 106 TCID50/mL (0.94 sensitivity, 0.96 specificity and 95% diagnostic accuracy). The performance of this chemiluminescence immunoassay would not permit it to replace molecular testing for diagnosing SARS-CoV-2, but may enable rapid and efficient detection of subjects with high SARS-CoV-2 viral load, who are responsible for the largest proportion of infectious clusters.


Ribosomes: analysis by cesium sulfate gradient centrifugation.
Ribosomes from the cytoplasm of a mammalian tissue, HeLa cells, were partially purified and then concentrated into a zone by prolonged centrifugation in Cs(2)SO(4) solution (density, 1.43 grams per milliliter). Ribosomal preparations treated with bentonite prior to density-gradient centrifugation showed single peaks after centrifugation. Material recovered from the peaks represented aggregates of the original 74S particle.


Recovery of endemic dragonflies after removal of invasive alien trees.
Because dragonflies are very sensitive to alien trees, we assessed their response to large-scale restoration of riparian corridors. We compared three types of disturbance regime--alien invaded, cleared of alien vegetation, and natural vegetation (control)--and recorded data on 22 environmental variables. The most significant variables in determining dragonfly assemblages were percentage of bank cover and tree canopy cover, which indicates the importance of vegetation architecture for these dragonflies. This finding suggests that it is important to restore appropriate marginal vegetation and sunlight conditions. Recovery of dragonfly assemblages after the clearing of alien trees was substantial. Species richness and abundance at restored sites matched those at control sites. Dragonfly assemblage patterns reflected vegetation succession. Thus, initially eurytopic, widespread species were the main beneficiaries of the removal of alien trees, and stenotopic, endemic species appeared after indigenous vegetation recovered over time. Important indicator species were the two national endemics (Allocnemis leucosticta and Pseudagrion furcigerum), which, along with vegetation type, can be used to monitor return of overall integrity of riparian ecology and to make management decisions. Endemic species as a whole responded positively to restoration, which suggests that indigenous vegetation recovery has major benefits for irreplaceable and widespread generalist species.


Assessing Changes in Surgical Site Infections and Antibiotic Use among Caesarean Section and Herniorrhaphy Patients at a Regional Hospital in Sierra Leone Following Operational Research in 2021.
Surgical site infections (SSIs) are a major public health threat to the success of surgery. This study assessed changes in SSIs and use of antibiotics among caesarean section (CS) and herniorrhaphy patients at a regional hospital in Sierra Leone following operational research. This was a comparative before and after study using routine hospital data. The study included all the CS and herniorrhaphy patients who underwent surgery between two time periods. Of the seven recommendations made in the first study, only one concerning improving the hospital's records and information system was fully implemented. Three were partially implemented and three were not implemented. The study population in both studies showed similar socio-demographic characteristics. The use of postoperative antibiotics for herniorrhaphy in both studies remained the same, although a significant increase was found for both pre- and postoperative antibiotic use in the CS patients, 589/596 (98.8%) in 2023 and 417/599 (69.6%) in 2021 (p < 0.001). However, a significant decrease was observed in the overall incidence of SSIs, 22/777 (2.8%) in 2023 and 46/681 (6.7%) in 2021 (p < 0.001), and the incidence of SSIs among the CS patients, 15/596 (2.5%) in 2023 and 45/599 (7.5%) in 2021 (p < 0.001). The second study highlights the potential value of timely assessment of the implementation of recommendations following operational research.


A survey of knowledge, attitudes, and beliefs of medical students concerning antimicrobial use and resistance.
Physicians who are insufficiently prepared to make choices on antibiotic selection may use antibiotics inappropriately. We surveyed medical students' perceptions and attitudes about their training on antimicrobial use to identify gaps in medical education. Medical students at an urban medical school in the northeast were e-mailed a link to an online survey. The survey was online for 1 week, after which time the survey responses were downloaded and analyzed. Thirty percent of medical students responded to the survey (n = 304). The majority of third- and fourth-year medical students believe that antibiotics are overused in the hospital and in outpatient areas. Over three quarters of the students would like more education on antibiotic selection, and 83% wanted this education to be during the third year of medical school. The resources they used the most for antibiotic selection included other physicians and handheld programs such as Epocrates, but no clear resource emerged as the dominant preference. Medical students recognized the importance of judicious antibiotic use and would like greater instruction on how to choose antibiotics appropriately. Medical school curricula should be expanded in the third year of medical school to provide students with additional training timed with their clinical rotations.


Molecular characterization and detection of mutations associated with resistance to succinate dehydrogenase-inhibiting fungicides in Alternaria solani.
Early blight, caused by Alternaria solani, is an economically important foliar disease of potato in several production areas of the United States. Few potato cultivars possess resistance to early blight; therefore, the application of fungicides is the primary means of achieving disease control. Previous work in our laboratory reported resistance to the succinate dehydrogenase-inhibiting (SDHI) fungicide boscalid in this plant pathogen with a concomitant loss of disease control. Two phenotypes were detected, one in which A. solani isolates were moderately resistant to boscalid, the other in which isolates were highly resistant to the fungicide. Resistance in other fungal plant pathogens to SDHI fungicides is known to occur due to amino acid exchanges in the soluble subunit succinate dehydrogenase B (SdhB), C (SdhC), and D (SdhD) proteins. In this study, the AsSdhB, AsSdhC, and AsSdhD genes were analyzed and compared in sensitive (50% effective concentration [EC50] < 5 μg ml(-1)), moderately resistant (EC50 = 5.1 to 20 μg ml(-1)), highly resistant (EC50 = 20.1 to 100 μg ml(-1)), and very highly resistant (EC50 > 100 μg ml(-1)) A. solani isolates. In total, five mutations were detected, two in each of the AsSdhB and AsSdhD genes and one in the AsSdhC gene. The sequencing of AsSdhB elucidated point mutations cytosine (C) to thymine (T) at nucleotide 990 and adenine (A) to guanine (G) at nucleotide 991, leading to an exchange from histidine to tyrosine (H278Y) or arginine (H278R), respectively, at codon 278. The H278R exchange was detected in 4 of 10 A. solani isolates moderately resistant to boscalid, exhibiting EC50 values of 6 to 8 μg ml(-1). Further genetic analysis also confirmed this mutation in isolates with high and very high EC50 values for boscalid of 28 to 500 μg ml(-1). Subsequent sequencing of AsSdhC and AsSdhD genes confirmed the presence of additional mutations from A to G at nucleotide position 490 in AsSdhC and at nucleotide position 398 in the AsSdhD, conferring H134R and H133R exchanges in AsSdhC and AsSdhD, respectively. The H134R exchange in AsSdhC was observed in A. solani isolates with sensitive, moderate, highly resistant, and very highly resistant boscalid phenotypes, and the AsSdhD H133R exchange was observed in isolates with both moderate and very high EC50 value boscalid phenotypes. Detection and differentiation of point mutations in AsSdhB resulting in H278R and H278Y exchanges in the AsSdhB subunit were facilitated by the development of a mismatch amplification mutation assay. Detection of these two mutations in boscalid-resistant isolates, in addition to mutations in AsSdhC and AsSdhD resulting in an H134R and H133R exchange, respectively, was achieved by the development of a multiplex polymerase chain reaction to detect and differentiate the sensitive and resistant isolates based on the single-nucleotide polymorphisms present in all three genes. A single A. solani isolate with resistance to boscalid did not contain any of the above-mentioned exchanges but did contain a substitution of aspartate to glutamic acid at amino acid position 123 (D123E) in the AsSdhD subunit. Among A. solani isolates possessing resistance to boscalid, point mutations in AsSdhB were more frequently detected than mutations in genes coding for any other subunit.


Cut, via CrebA, transcriptionally regulates the COPII secretory pathway to direct dendrite development in Drosophila.
Dendrite development is crucial in the formation of functional neural networks. Recent studies have provided insights into the involvement of secretory transport in dendritogenesis, raising the question of how the secretory pathway is controlled to direct dendritic elaboration. Here, we identify a functional link between transcriptional regulatory programs and the COPII secretory machinery in driving dendrite morphogenesis in Drosophila dendritic arborization (da) sensory neurons. MARCM analyses and gain-of-function studies reveal cell-autonomous requirements for the COPII coat protein Sec31 in mediating da neuron dendritic homeostasis. We demonstrate that the homeodomain protein Cut transcriptionally regulates Sec31 in addition to other components of COPII secretory transport, to promote dendrite elaboration, accompanied by increased satellite secretory endoplasmic reticulum (ER) and Golgi outposts primarily localized to dendritic branch points. We further establish a novel functional role for the transcription factor CrebA in regulating dendrite development and show that Cut initiates a gene expression cascade through CrebA that coordinately affects the COPII machinery to mediate dendritic morphology.


Angiopoietin-2 is expressed in oral Kaposi's sarcoma.
Kaposi's sarcoma (KS) persists today as a highly prevalent vascular cancer, often found in HIV patients. Studies have shown that angiopoietin 2 (Ang2), a pro-angiogenic protein, is involved in the pathogenesis of this tumor. However, expression of this protein has not been investigated in oral KS lesions. Thus, we aimed to investigate the expression of Ang2 in samples of oral KS. Immunohistochemistry was used to evaluate Ang2 expression in 14 oral KS cases, with degrees of expression being analyzed in a semi-quantitative manner. In addition, clinical information such as age, gender, race, tumor location, size, color, and appearance, as well as HIV status, was collected and included in the analysis. All patients were white males, mostly HIV-positive, with a mean age of 40 years. Clinically, the lesions were dark red/blue/purple masses, ranging from 1 to 2.5 cm in diameter, found in various locations such as the tongue, palate, and gingiva. Expression of Ang2 was noted in 72% (10/14) of the samples. Of these, 10% showed weak expression, 60% moderate, and 30% strong expression. Our results indicate that Ang2 is expressed in oral KS and, consistent with results from previous studies, show that Ang2 may contribute to the pathogenesis of this lesion.


Chronic middle ear effusions. Immunochemical and bacteriological investigations.
Middle ear effusions from 100 patients (ages 6 months to 10 years) with serous otitis media were examined. The IgA, IgG, and lysozyme were demonstrated at a higher level in the effusions than the corresponding sera, indicating local production. The mucoid type contained higher level of immunoglobulins and lysozyme compared to serous type effusions. Bacteria were found in 77 percent of the effusions by means of a smear, and 52 percent yielded positive bacterial culture. The incidence of positive culture in effusions of the patients less than 6 years of age was 60 percent, while the group older than 6 years old was 32%, and the group over 8 was only 22 percent. Bacterial recovery rate was inversely related to the dramatic increase with age of IgA and IgG and lysozyme levels in effusions.


Advances in cardiac imaging.
Silent myocardial ischemia is the most important target of a variety of imaging modalities. The ultimate technology would include anatomic, functional, and myocardial perfusion information. Ultrafast computer tomography, as this time, is the best modality for studying the coronary arteries. Positron Emission Tomography remains the gold standard for studies of myocardial blood flow and myocardial utilization.


Longitudinal Memory Profiles in Behavioral-Variant Frontotemporal Dementia and Alzheimer's Disease.
Alzheimer's disease (AD) and behavioral-variant of frontotemporal dementia (bvFTD) can present with an overlapping neuropsychological profile, which often hinders their clinical differentiation. To compare changes over time in memory, general cognition tasks, and functional scales between bvFTD and AD. Consecutive cases diagnosed with probable bvFTD (n = 22) and typical AD (n = 31) with at least two clinical visits were selected. Of these, 13 (9 AD, 4 bvFTD) underwent Pittsburgh compound B PET scan, which supported the clinical diagnosis in all cases. Mixed-model regressions were used to estimate the differential rate of decline on selected tasks between cohorts. Analyses demonstrated that, despite equivalent baseline performance, bvFTD patients experienced a more rapid functional deterioration and a steeper decline in global cognition than AD patients. At baseline, both groups were impaired on executive function and memory tasks compared to controls, but these deficits were more marked in the bvFTD group. In addition, performance on these domains continued to decline more rapidly in this group. Neither the initial neuropsychological assessment nor projected performances can reliably distinguish the totality of bvFTD and AD individuals. Nevertheless, annual rates of progression on cognitive tasks provide valuable information and will potentially help establish the impact of future therapeutic treatments in these dementia syndromes.


Clinical findings among sexually abused children with and without developmental disabilities.
Although children with developmental disabilities have a heightened risk for sexual abuse compared to those without disabilities, little is known about effects of sexual abuse on this population. Some researchers suggest that the effects are similar, and victims appear to share the same range of variability as people without disabilities. Others suggest that the effects may be complicated by limited coping resources and disability-related issues. Documented effects of sexual abuse on individuals with developmental disabilities was discussed. Clinical findings among a group of 43 children with and 43 children without developmental disabilities referred for treatment of child sexual abuse were also presented.


Global age-dependent differences in gene expression in response to calvarial injury.
Children less than 2 years of age are capable of healing large calvarial defects, whereas adults have been found to lack this endogenous ability. In this study, we used microarray analysis to compare genomewide expression patterns during active regeneration after injury with calvaria in skeletally immature and mature mice. Parietal bone defects were created in 6-day-old (juvenile) and 60-day-old (adult) mice using a 4-mm trephine bit (n = 20 mice per age group). The calvarial disc was removed, leaving the underlying dura mater intact. Two weeks after injury, the region of regeneration with the underlying dura mater was harvested, and RNA was extracted for microarray analysis. The 25 most differentially upregulated genes in juvenile regenerates compared with adults were listed, as well as selected bone-related genes. In addition, QRT-PCR confirmation of specific genes was performed for validation. Juvenile regenerates expressed significantly greater amounts of BMP-2, -4, -7, as well as FGF-2 and its receptor FGFR-1. Various other growth factors were also noted to be upregulated, including IGF-2 and Ptn. This corresponded with the increased expression of markers for osteogenic differentiation of Sparc and Oc. Markers of osteoclast activity, Acp5, Ctsk, and Mmp2, were noted to be greater in juvenile regenerates compared with adults. The observation of Mmp14 upregulation, however, highlights the importance of balanced osteoclast-mediated bone resorption for ultimate healing. The 2 most differentially regulated genes, transthyretin (Ttr) and prostaglandin D2 synthase (Ptgds), highlight the potential role of retinoic acid signaling and the prostaglandin axis on skeletal regeneration. These findings underscore the multitude of biomolecular mechanisms at play, allowing juvenile calvaria to heal after injury. The identification of various growth factors and cytokines involved also suggests novel therapeutic strategies for tissue-engineering purposes.


A second case of diphyllobothriasis in Malaysia.
Diphyllobothriasis was first reported in Malaysia in 2002. We are reporting a second case of diphyllobothriasis in Malaysia. The patient was a 37-year-old Chinese male seen at the outpatient clinic with a complaint of passing strands of white color flat worm in his stool. He had no other abdominal complaints. Laboratory and physical examinations were normal. Diphyllobothrium latum was confirmed by examination of the gravid proglottids passed out and the typical operculated eggs expelled from the ruptured proglottids. The patient had a history of eating raw fish. He was treated with a single dose of praziquantel.


Inhibition of tumor promotion in mouse skin by 1 alpha,25-dihydroxyvitamin D3.
1 alpha,25-Dihydroxyvitamin D3 [1 alpha,25(OH)2D3], a hormonally active form of vitamin D3, was found to inhibit the promotional phase of 7,12-dimethylbenz[a]anthracene-induced skin carcinogenesis in female Sencar mice. Topical application of 1 alpha,25-(OH)2D3 once a week at a dose of 1 micrograms or less, a tolerable dose from hypercalcemia, dose dependently inhibited tumor promotion by 12-O-tetradecanoylphorbol-13-acetate (TPA). When 1 micrograms of 1 alpha,25(OH)2D3 was applied 30 min before 5 micrograms of TPA, the times required for 50 and 100% tumor incidence were delayed about 2.5 and 7 weeks, respectively, and the number of tumors per mouse was decreased by 25-30%. This inhibitory effect was more pronounced when examined by a two-stage promotion protocol, in which a single application of 5 micrograms of TPA (Stage I) was followed by repeated applications of 5 micrograms of mezerein once a week for 19 weeks (Stage II). When 1 alpha,25(OH)2D3 at 1 micrograms was applied at Stage I + II or Stage II, tumor formation was markedly suppressed, resulting in decrease of about 70-80% in the incidence and 87-90% in the number of tumors per mouse. Application of 1 alpha,25(OH)2D3 at Stage I only did not inhibit tumor formation, indicating that 1 alpha,25(OH)2D3 specifically inhibited Stage II promotion. These results are in good agreement with the previous and present findings that 1 alpha,25(OH)2D3 inhibited induction of epidermal ornithine decarboxylase by TPA and mezerein. The possibility that 1 alpha,25(OH)2D3 suppressed tumor promotion by killing initiated cells rather than inhibiting promotion was ruled out by an experiment in which TPA was applied to the 1 alpha,25(OH)2D3 alone-treated animals.


Engineering attenuated virus vaccines by controlling replication fidelity.
Long-lasting protection against viral infection is best achieved by vaccination with attenuated viruses. Obtaining stably attenuated vaccine strains has traditionally been an empirical process, which greatly restricts the number of effective vaccines for viral diseases. Here we describe a rational approach for engineering stably attenuated viruses that can serve as safe and effective vaccines. Our approach exploits the observation that restricting viral population diversity by increasing replication fidelity greatly reduces viral tissue tropism and pathogenicity. We show that poliovirus variants with reduced genetic diversity elicit a protective immune response in an animal model of infection. Indeed, these novel vaccine candidates are comparable in efficacy to the currently available Sabin type 1 vaccine strain, but have the added advantage of being more stable, as their increased replication fidelity prevents reversion to the pathogenic wild-type phenotype. We propose that restricting viral quasispecies diversity provides a general approach for the rational design of stable, attenuated vaccines for a wide variety of viruses.


Hsp28stl: a P-element insertion mutation that alters the expression of a heat shock gene in Drosophila melanogaster.
We have identified and cloned a mutant allele of the small heat shock gene Hsp28 of Drosophila melanogaster. This allele, which we have called Hsp28stl, produces small amounts of a single aberrantly large, heat-inducible transcript in heat-shocked flies, while a normal-sized Hsp28 transcript is present only in fertile females. No Hsp28 transcript at all is detected in mutant prepupae, a stage when wildtype flies show high levels of Hsp28 RNA. We have cloned the Hsp28stl allele, and have found that a 1.3-kb defective P-element is present 5' to Hsp28 in the mutant line. The site of P-element insertion lies between the Hsp28 "TATA box" sequence and the Hsp28 RNA cap site; in contrast to previously described P-element insertions, the element at Hsp28stl is flanked by a two base pair duplication of the insertional target sequence. The results suggest that this insert may separate elements regulating heat-inducible and developmental expression of Hsp28, leading to the different patterns of transcription observed.


Common genetic variation in the autoimmune regulator (AIRE) locus is associated with autoimmune Addison's disease in Sweden.
Autoimmune Addison's disease (AAD) is the predominating cause of primary adrenal failure. Despite its high heritability, the rarity of disease has long made candidate-gene studies the only feasible methodology for genetic studies. Here we conducted a comprehensive reinvestigation of suggested AAD risk loci and more than 1800 candidate genes with associated regulatory elements in 479 patients with AAD and 2394 controls. Our analysis enabled us to replicate many risk variants, but several other previously suggested risk variants failed confirmation. By exploring the full set of 1800 candidate genes, we further identified common variation in the autoimmune regulator (AIRE) as a novel risk locus associated to sporadic AAD in our study. Our findings not only confirm that multiple loci are associated with disease risk, but also show to what extent the multiple risk loci jointly associate to AAD. In total, risk loci discovered to date only explain about 7% of variance in liability to AAD in our study population.


Non-reproductive effects of sex steroids: their immunoregulatory role.
In this special issue of Current Topics in Medicinal Chemistry, the reader will find reviewed some of the hottest topics in the field of the non-reproductive effects of sex steroids. Cabrera-Muñoz et al., show that progesterone participates in the regulation of human brain tumors growth. The contribution of Martocchia suggests that sex steroid receptor modulating drugs provide new therapeutic approaches to autoimmune diseases. The role of sex steroid participation in the differentiation of stem cells to neurones is discussed by I. Velasco. Pérez-Torres and collaborators demonstrate that sex steroids play an important role in the appearance and development of renal diseases and the metabolic syndrome, the new epidemics of our century. Paris and Frye hypthetize that gestational stress, have effects on cognitive performance and/or neuronal integrity in the fetus, and that exposure to variable stress during gestation can perturb cognitive performance, concomitant with dendrite development in hippocampus and diencephalon. Muñoz-Cruz et al. thoroughly review the growing body of evidence that shows reciprocal relationship between sex steroids and the immune system, and conclude that understanding the mechanisms of action of sex steroids on immune cells is important for further progress in the development of novel therapies for chronic diseases associated to immune dysregulation. Besides, the effects of sexual steroids on pancreatic function and diabetes are reviewed by Morimoto et al. Yanes et al. review some of the contradictions raised in the context of the recently proposed critical period hypothesis, which takes into account the frame-time after cessation of ovarian function. Finally, another vey intetresting aspect of the non-reproductive effects of sex-steroids, is the related to some cognition-related aspects, which is reviewed by Picazo et al.


Natural killer cell activity in families of patients with systemic lupus erythematosus: demonstration of a killing defect in patients.
Natural killer (NK) cell cytotoxic activity and cell frequency, expressed as a percentage of total lymphocytes, have been determined in peripheral blood mononuclear cells from first-degree relatives of patients with systemic lupus erythematosus (SLE), the patients themselves, a group of rheumatoid arthritis (RA) patients and controls. Low levels of killing activity relative to controls were found in some members of all groups with the extent of depression falling into two ranges. Moderate reductions were seen in female (3/31, 10%) and male (4/14, 29%) relatives of SLE patients, female (12/60, 20%) and male (3/4, 75%) SLE patients and female RA patients (6/17, 35%). A more profound depression of killing activity was confined to other female SLE patients (15/60, 25%). There were strong correlations in all groups between killing activity and percentage of NK cells, but analysis of the ratio of these parameters and studies with purified preparations of NK cells suggest that the reduced activity in SLE frequently involves a defect in the killing capacity of the individual cells in addition to the reduced levels of NK cells. Azathioprine (AZA), which was used in treatment of 12 SLE patients, was invariably associated with low values of killing activity. It appears to substantially reduce the percentage of NK and B cells in an action unconnected with the NK cell abnormalities associated with SLE. The finding of low killing activity in relatives and a correlation between their activity and that of their patients support the view that NK cell deficiency is a genetic determinant of SLE. NK cells in SLE may produce insufficient levels of cytokines required for the regulation of IgG production.


Rocky Mountain spotted fever in Mexico: past, present, and future.
Rocky Mountain spotted fever, a tick-borne zoonosis caused by Rickettsia rickettsii, is among the most lethal of all infectious diseases in the Americas. In Mexico, the disease was first described during the early 1940s by scientists who carefully documented specific environmental determinants responsible for devastating outbreaks in several communities in the states of Sinaloa, Sonora, Durango, and Coahuila. These investigators also described the pivotal roles of domesticated dogs and Rhipicephalus sanguineus sensu lato (brown dog ticks) as drivers of epidemic levels of Rocky Mountain spotted fever. After several decades of quiescence, the disease re-emerged in Sonora and Baja California during the early 21st century, driven by the same environmental circumstances that perpetuated outbreaks in Mexico during the 1940s. This Review explores the history of Rocky Mountain spotted fever in Mexico, current epidemiology, and the multiple clinical, economic, and social challenges that must be considered in the control and prevention of this life-threatening illness.


Regulation of the effector stages of experimental autoimmune encephalomyelitis via neuroantigen-specific tolerance induction. III. A role for anergy/deletion.
Our previous work has shown that specific peripheral immune tolerance induced by the intravenous administration of ECDI-fixed, antigen-coupled syngeneic splenocytes is an extremely efficient method for prevention and treatment of chronic relapsing experimental autoimmune encephalomyelitis (R-EAE) in susceptible SJL/J mice. The current study examined the mechanisms by which unresponsiveness is induced in primed encephalitogenic T cells. The results indicate that the inhibition of MBP-specific T cells by the i.v. injection of MBP-coupled splenocytes is not due to the induction of antigen-specific regulatory T cells, but rather to the induction of anergy/deletion of the effector cells. This conclusion is supported by the findings that spleen or lymph node cells isolated from MBP-tolerant mice fail to inhibit the adoptive transfer of R-EAE in cotransfer assays, and that tolerance is not inhibited by prior thymectomy or prior treatment with cyclophosphamide or anti-CD8 monoclonal antibody. In contrast, we demonstrate that splenocytes from MBP-tolerized, asymptomatic mice have a significantly reduced ability to serially transfer R-EAE to naive secondary recipients following antigen re-activation in vitro, in the first several weeks following tolerization, but that the ability to serially transfer R-EAE returns to sham tolerant control levels within 1-2 months. We also demonstrate a significantly reduced precursor frequency of MBP-specific, IL-2-producing T cells in the MBP-tolerant within three days of treatment. Collectively, the data most closely support a model wherein inhibition of MBP-specific encephalitogenic CD4+ effector T cells by i.v. injected MBP-coupled splenocytes is due to the direct induction of anergy/deletion from which they can recover over time.


RNA-binding proteins with prion-like domains in ALS and FTLD-U.
Amyotrophic lateral sclerosis (ALS, also known as Lou Gehrig's disease) is a debilitating, and universally fatal, neurodegenerative disease that devastates upper and lower motor neurons. The causes of ALS are poorly understood. A central role for RNA-binding proteins and RNA metabolism in ALS has recently emerged. The RNA-binding proteins, TDP-43 and FUS, are principal components of cytoplasmic inclusions found in motor neurons of ALS patients and mutations in TDP-43 and FUS are linked to familial and sporadic ALS. Pathology and genetics also connect TDP-43 and FUS with frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U). It was unknown whether mechanisms of FUS aggregation and toxicity were similar or different to those of TDP-43. To address this issue, we have employed yeast models and pure protein biochemistry to define mechanisms underlying TDP-43 and FUS aggregation and toxicity, and to identify genetic modifiers relevant for human disease. We have identified prion-like domains in FUS and TDP-43 and provide evidence that these domains are required for aggregation. Our studies have defined key similarities as well as important differences between the two proteins. Collectively, however, our findings lead us to suggest that FUS and TDP-43, though similar RNA-binding proteins, likely aggregate and confer disease phenotypes via distinct mechanisms.


TRAF6 regulates the effects of polarized maturation of tolerability: Marrow-derived dendritic cells on collagen-induced arthritis in mice.
The study aimed to investigate the relationship between tumor necrosis factor receptor-associated factor 6 (TRAF6) and a differentially mature dendritic cell (mDC) in collagen-induced arthritis (CIA) mice and to determine whether or not TRAF6 regulates the activation of an immature dendritic cell (iDC) and inhibits iDC maturation to induce immune tolerance. The mouse bone marrow stem cells were induced with recombinant granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and recombinant interleukin-4 (rmIL-4) to differentiate immature dendritic cells (DCs), which were divided into four groups with different maturation states: rmGM-CSF, rmIL-4; TNF-α; LPS; and FK506 group. The levels of the cell surfaces of CD80, CD86, and MHI-II were analyzed by flow cytometry to prove DCs at different levels of maturity. The expression of IL-12 in DCs at different maturation states was detected by enzyme-linked immunosorbent assay (ELISA). The expression of TRAF6 mRNA and protein in each group of DCs was detected by a reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis. The results revealed that the differentiation of bone marrow cells into iDCs was significantly induced by cytokines (rmGM-CSF, IL-4). CD80, CD86, MHC-II were expressed in the four groups, and the difference between them was statistically significant (P<0.05). A higher degree of DC differentiation led to a gradual increase of IL-12 secretion in the four groups. The difference was statistically significant (P<0.05) for this secretion (group D, 10,620.73±276.73 pg/ml). The expression levels of TRAF6 mRNA were significantly higher in group D than those in the other three groups (P<0.01). Although there was no significant difference in the expression levels of TRAF6 mRNA between groups B and C, the expression levels of TRAF6 mRNA between groups B and C were higher than those of the control group. The TRAF6 protein expression was higher in group D than that in the other three groups (P<0.01), and the difference was statistically significant. There was a statistically significant difference in the TRAF6 protein expression between group A and groups B and C, but the expression in group C was higher than that in group B (P<0.01). In conclusion, the expression of co-stimulatory molecules gradually increased in the DCs of different maturation states, and the expression of IL-12, TRAF6 mRNA, and TRAF6 protein positively correlated with the degree of DC maturation. TRAF6 is important in iDC polarity and maturation.


N-Heterocyclic carbenes and their precursors in functionalised porous materials.
Though N-heterocyclic carbenes (NHCs) have emerged as diverse and powerful discrete functional molecules in pharmaceutics, nanotechnology, and catalysis over decades, the heterogenization of NHCs and their precursors for broader applications in porous materials, like metal-organic frameworks (MOFs), porous coordination polymers (PCPs), covalent-organic frameworks (COFs), porous organic polymers (POPs), and porous organometallic cages (POMCs) was not extensively studied until the last ten years. By de novo or post-synthetic modification (PSM) methods, myriads of NHCs and their precursors containing building blocks were designed and integrated into MOFs, PCPs, COFs, POPs and POMCs to form various structures and porosities. Functionalisation with NHCs and their precursors significantly expands the scope of the potential applications of porous materials by tuning the pore surface chemical/physical properties, providing active sites for binding guest molecules and substrates and realizing recyclability. In this review, we summarise and discuss the recent progress on the synthetic methods, structural features, and promising applications of NHCs and their precursors in functionalised porous materials. At the end, a brief perspective on the encouraging future prospects and challenges in this contemporary field is presented. This review will serve as a guide for researchers to design and synthesize more novel porous materials functionalised with NHCs and their precursors.


Adolescent, Parent, and Provider Perspectives of a Social-Media-Based Support Tool for Parents of Teens in Treatment for Substance Use.
Adolescent substance use disorder contributes to negative psychosocial and physical health outcomes. Parents best support their child's treatment and recovery when they are equipped with knowledge and skills; however, barriers hinder parental engagement. Social-media-based groups present an opportunity for parents who may not otherwise seek support. Qualitative data were collected via focus groups to examine the opinions of adolescents, parents, and treatment providers on the potential utility and possible concerns of a Facebook-based parent support group; input constituted the development phase for an online group (PURPOSE). Fifty-eight participants, recruited through adolescent treatment programs, took part in six focus groups: two parent ( n = 18), two adolescent ( n = 21), and two provider ( n = 19) groups. Groups were audio recorded and transcribed, and themes were extracted. Three main themes emerged: (a) traditional support groups and treatment services, (b) PURPOSE intervention content and delivery, and (c) issues related to PURPOSE implementation. Traditional treatment/support groups can be helpful, but participation in them is limited. Emotional support offered by peers is valuable, but parents also need and seek out psychoeducation, parenting skills, and self-care support. PURPOSE could offer easy access to skills and real-time peer support.


The effect of promoting intrauterine attachment in primiparas on postdelivery attachment.
Twenty-two women participated in a modified replicate study which examined the effects of prenatal maternal attachment behaviors during first pregnancies. A pretest reflected the degree of maternal-fetal attachment behaviors and attitudes already formed. The women in the experimental group received three interventions aimed at directing the mothers' attentions toward their fetuses. The remaining women received routine prenatal care only. All mothers were observed once in the first few days postpartum to assess maternal attachment behaviors. Data analysis did not substantiate the findings of the earlier study. Factors identified in the added pretest may have accounted for this difference.


The effects of Baduanjin exercise on the subjective memory complaint of older adults: A randomized controlled trial.
This study aimed to explore the efficacy of Baduanjin exercise on promoting memory function, executive function and general self-efficacy, decreasing the level of subjective memory complaints of older adults.In this randomized controlled trial, 80 patients were randomly allocated in a 1:1 ratio to 12-week Baduanjin exercise group or 12-week control group. Subjective memory complaint questionnaire, Auditory verbal learning test, Trail Making Test and General Self-Efficacy Scale was used to assess the subjective memory complaint level, memory function, executive function and general self-efficacy level at three times (baseline, after intervention and follow up at 3 months). One-way repeated measures analysis of variance was used to compare the outcome variables of the two groups.There were no significant differences between the Baduanjin exercise and the control group at baseline in demographic, SMCQ, MoCA, and GDS-15. Compared to participants in the control group, participants in the Baduanjin group had a significantly improvement in memory function (F = 46.93, P < .00), executive function (F = 317.83, P < .00) and general self-efficacy (F = 38.72, P < .00) at the end of 12-week intervention period and after 3months follow-up period (P < .01). At the same time, participants in the Baduanjin group had a significantly greater decrease in subjective memory complaint scores at the end of 12-week intervention period and after 3months follow-up period (F = 24.53, P < 0.00).Baduanjin exercise appears to be a feasible and acceptable intervention to improve subjective memory complaint among older adults.


Involvement of purinergic nerves in the NANC inhibitory junction potentials in pigeon oesophageal smooth muscle.
1. Electrical field stimulation (EFS) (0.5 ms in train of 2-32 Hz for 300 ms) in smooth muscle of pigeon oesophagus, in the presence of atropine (1 microm) and guanethidine (1 microm), elicited an inhibitory response consisting of a transient hyperpolarization (inhibitory junction potential, IJP) associated with muscle relaxation. 2. Sodium nitroprusside (SNP, 100 microm) induced hyperpolarization correlated to mechanical relaxation. 3. The nitric oxide (NO) synthase inhibitor N(omega)-nitro-l-arginine (from 0.1 to 100 microm) caused a concentration-dependent reduction of electromechanical response to EFS indicating a role for NO in this response. 4. Apamin (1 microm) reduced both IJP and relaxation to EFS but was without effect on the response to SNP indicating a role for purines, which are also blocked by apamin. 5. Adenosine, AMP, ADP and ATP (all from 1 microm to 1 mm) application caused transient hyperpolarization and muscular relaxation with the following order of potency: adenosine > AMP > ADP > ATP. 6. Inhibitory responses evoked by purines are TTX (1 microm) insensitive but they were inhibited by apamin. This indicates that a purine component for the non-adrenergic non-cholinergic (NANC) response exists but the purine receptor site is not located on the neurone. 7. Overall these results suggest that NANC inhibitory response elicited by EFS presents two different components apamin-sensitive, probably purines-mediated and apamin-insensitive probably NO-mediated as apamin only partially block the response to EFS.


Critical physiotherapy: a ten-year retrospective.
Critical physiotherapy has been a rapidly expanding field over the last decade and could now justifiably be called a professional sub-discipline. In this paper we define three different but somewhat interconnected critical positions that have emerged over the last decade that share a critique of physiotherapy's historical approach to health and illness, while also diverging in the possibilities for new forms of practice and thinking. These three positions broadly align with three distinctive philosophies: approaches that emphasize lived experience, social theory, and a range of philosophies increasingly referred to as the "posts". In this paper we discuss the origins of these approaches, exploring the ways they critique contemporary physiotherapy thinking and practice. We offer an overview of the key principles of each approach and, for each in turn, suggest readings from key authors. We conclude each section by discussing the limits of these various approaches, but also indicate ways in which they might inform future thinking and practice. We end the paper by arguing that the various approaches that now fall under the rubric of critical physiotherapy represent some of the most exciting and opportune ways we might (re)think the future for the physiotherapy profession and the physical therapies more generally.


Patients as Rights Holders.
Autonomy and consent have been central values in Western moral and political thought for centuries. One way of understanding the bioethical models that started to develop, especially in the 1970s, is that they were about the fusion of a long-standing professional ethics with the core values underpinning modern political institutions. That there was a need for this kind of fusion is difficult to dispute, especially since the provision of health care has in most developed countries become an ever more important concern of our political institutions, with governments playing a significant role in regulating and facilitating the provision of health care and in many countries even largely organizing it. There is, nevertheless, still room for dispute about how best to achieve this fusion and how to best think about autonomy and consent in a biomedical context. The simplest model we can have is probably about how being a person is largely about having the capacity of autonomous choice and that the main mode through which we exercise autonomy is by providing informed consent. Yet, liberal democracy's core idea that human beings have a high and equal value is also found in other accounts of the person. The human-rights framework provides an alternative model for thinking about personhood and about patient care. The human-rights approach is grounded, not in an account of autonomy (although it has something to say about autonomy), but in an account of the moral and political personhood that people possess merely by being human beings. In this approach, values like dignity and integrity, both highly relevant in a bioethical context, are identified as distinct values rather than being derived from and therefore reduced to respect for autonomous choice. The human-rights approach can supplement the problematic notion of autonomy that has been central to bioethics by placing this notion in a broader, strongly pluralistic framework.


Molecular engineering of a high quantum yield NIR-II molecular fluorophore with aggregation-induced emission (AIE) characteristics for in vivo imaging.
NIR-II biological imaging (1000-1700 nm) has shown promise for deep tissue penetration, high spatial resolution, and low background noise. Among all the NIR-II probes, organic probes particularly attract huge attention due to their excellent stability and biocompatibility, which have the most potential for clinical translation. However, most previously reported organic NIR-II fluorescent agents often suffer from low quantum yields in aqueous solution. Herein, we developed a novel D-π-A-π-D-type NIR II chromophore XA1 with AIE characteristics based on a new design strategy for NIR-II AIE fluorophores. Owing to their intrinsic aggregation-induced emission enhancement nature, the formulated XA1 NPs show a high fluorescence quantum yield up to 14.8%, which is higher than those of most previously reported organic NIR-II fluorophores. Based on the XA1 NPs, noninvasive imaging of limb and cerebral vessels is achieved with a high signal-to-background ratio and deep penetration. Furthermore, the XA1 NPs can be used as good contrast agents for high resolution imaging of blood vessels of tumors and precise detection of tumors based on the EPR mechanism. Collectively, our work demonstrated a novel strategy for designing and manufacturing NIR-II fluorophores with AIE characteristics and proved that XA1 NPs are highly promising NIR-II probes for biomedical imaging under physiological and pathological conditions.


Association Between Lower Trapezius Isometric Strength and Y-Balance Test Upper Quarter Performance in College Volleyball Players.
Knowing the possible association between the isometric strength of the shoulder rotators, scapular muscles, and the Y-balance test upper quarter (YBT-UQ) performance could help identify which indicators of shoulder stability should be considered in this field test. This study aimed to determine whether the isometric strength of the shoulder rotators and scapular muscles is associated with the YBT-UQ performance of the dominant upper limb in amateur volleyball players. Cross-sectional study. A convenience sample of 22 male and 18 female volleyball players (≥12 h of training/week) between 18 and 26 years of age. The isometric strength of the middle trapezius, lower trapezius, serratus anterior, internal, and external rotator muscles was assessed with a handheld dynamometer. Participants performed the YBT-UQ in the superolateral, medial, and inferolateral directions. The absolute isometric peak force (in Newtons) was normalized to body weight (in Newtons per kilogram) for each muscle test. For each YBT-UQ direction, the distance (in centimeters) was normalized for upper limb length (in percentage). A backward multiple linear regression analysis was used to determine the associations between variables. The analysis revealed that the isometric strength of the lower trapezius (β = 26.82; 95% confidence interval, 21.24-32.40) is associated with inferolateral YBT-UQ performance (adjusted R2 = .706; P < .001). This factor explains 70% of the variability of the YBT-UQ in the inferolateral direction. Lower trapezius isometric strength is associated with inferolateral YBT-UQ performance of the dominant upper limb in amateur volleyball players. These findings could help in the development of more specific training programs and rehabilitation goals according to the performance of the athletes in the test.


Experimental comparison of relative RT-qPCR quantification approaches for gene expression studies in poplar.
RT-qPCR is a powerful tool for analysing gene expression. It depends on measuring the increase in fluorescence emitted by a DNA-specific dye during the PCR reaction. For relative quantification, where the expression of a target gene is measured in relation to one or multiple reference genes, various mathematical approaches are published. The results of relative quantification can be considerably influenced by the chosen method. We quantified gene expression of superoxide dismutase (SOD) and ascorbate peroxidase (APX) in the roots of two black poplar clones, 58-861 and Poli, which were subjected to drought stress. After proving the chosen reference genes actin (ACT), elongation factor 1 (EF1) and ubiquitin (UBQ) to be constantly expressed in the different watering regimes, we applied different approaches for relative quantification to the same raw fluorescence data. The results obtained using the comparative Cq method, LinRegPCR, qBase software and the Pfaffl model showed a good correlation, whereas calculation according to the Liu and Saint method produced highly variable results. However, it has been shown that the most reliable approach for calculation of the amplification efficiency is using the mean increase in fluorescence during PCR in each individual reaction. Accordingly, we could improve the quality of our results by applying the mean amplification efficiencies for each amplicon to the Liu and Saint method. As we could show that gene expression results can vary depending on the approach used for quantification, we recommend to carefully evaluate different quantification approaches before using them in studies analysing gene expression.


Effects of short-term isokinetic training with reciprocal knee extensors agonist and antagonist muscle actions: a controlled and randomized trial.
Previous studies have shown that preloading an antagonist muscle may increase the acute agonist neuromuscular performance. In addition, studies have suggested that very short-term resistance exercise (RE) programs may also be useful to increase strength and muscular performance. To evaluate the effects of three days of RE using a reciprocal action method on the muscular performance of healthy men and to compare these effects with those of a traditional RE group. Thirty-three men (21.1 ± 2.3 years) were randomly assigned to one of three groups: 1) reciprocal (REC; knee flexion immediately followed by a knee extension exercise); 2) traditional (TRA; non-preload; a concentric knee extension exercise); and 3) control (CON; no exercise). The REC and TRA subjects performed four sets of 10 repetitions at 60º/s with one minute of rest. The pre- and post-RE tests included two sets of four maximal concentric repetitions at 60º/s and 180º/s. A 3x2 ANOVA with Tukey post-hoc was used to analyze the differences in peak torque (PT), rating of acceleration development (RAD) and time to peak torque (TIMEtorque). A significant PT increase was found for REC and TRA (p<0.05) at 60º/s and for REC at 180º/s (p<0.05). There was a decrease in the RAD for REC and TRA (p<0.05), and TIMEtorque showed a significant decrease for REC. The inter-group analysis revealed that REC is more effective than TRA for PT gains at both velocities (p<0.05). It is recommended that REC offers benefits for the clinical practice of professionals involved in neuromuscular rehabilitation.


Controlling reactivity by remote protonation of a basic side group in a bifunctional photoacid.
6-Hydroxy-2-naphthoic acid and its sulfonate derivatives belong to a family of bifunctional photoacids where the -OH group acts as a proton donor and the -COO(-) group acts as a proton acceptor. Upon electronic excitation, the -OH group becomes more acidic and the -COO(-) group turns more basic. Change in the ionization state of one functional group causes a change (switch) in the reactivity of the other functional group. Using picosecond time-resolved and steady state spectroscopy, we find clear evidence for an ultrafast reactivity switch caused by a diffusional proton transfer through the water solvent between the two functional groups with no evidence of a concerted proton transfer.


Identification of regulatory networks in HSCs and their immediate progeny via integrated proteome, transcriptome, and DNA methylome analysis.
In this study, we present integrated quantitative proteome, transcriptome, and methylome analyses of hematopoietic stem cells (HSCs) and four multipotent progenitor (MPP) populations. From the characterization of more than 6,000 proteins, 27,000 transcripts, and 15,000 differentially methylated regions (DMRs), we identified coordinated changes associated with early differentiation steps. DMRs show continuous gain or loss of methylation during differentiation, and the overall change in DNA methylation correlates inversely with gene expression at key loci. Our data reveal the differential expression landscape of 493 transcription factors and 682 lncRNAs and highlight specific expression clusters operating in HSCs. We also found an unexpectedly dynamic pattern of transcript isoform regulation, suggesting a critical regulatory role during HSC differentiation, and a cell cycle/DNA repair signature associated with multipotency in MPP2 cells. This study provides a comprehensive genome-wide resource for the functional exploration of molecular, cellular, and epigenetic regulation at the top of the hematopoietic hierarchy.


Astrocytic TGF-β signaling limits inflammation and reduces neuronal damage during central nervous system Toxoplasma infection.
The balance between controlling infection and limiting inflammation is particularly precarious in the brain because of its unique vulnerability to the toxic effects of inflammation. Astrocytes have been implicated as key regulators of neuroinflammation in CNS infections, including infection with Toxoplasma gondii, a protozoan parasite that naturally establishes a chronic CNS infection in mice and humans. In CNS toxoplasmosis, astrocytes are critical to controlling parasite growth. They secrete proinflammatory cytokines and physically encircle parasites. However, the molecular mechanisms used by astrocytes to limit neuroinflammation during toxoplasmic encephalitis have not yet been identified. TGF-β signaling in astrocytes is of particular interest because TGF-β is universally upregulated during CNS infection and serves master regulatory and primarily anti-inflammatory functions. We report in this study that TGF-β signaling is activated in astrocytes during toxoplasmic encephalitis and that inhibition of astrocytic TGF-β signaling increases immune cell infiltration, uncouples proinflammatory cytokine and chemokine production from CNS parasite burden, and increases neuronal injury. Remarkably, we show that the effects of inhibiting astrocytic TGF-β signaling are independent of parasite burden and the ability of GFAP(+) astrocytes to physically encircle parasites.


[Waking up during general anaesthesia].
Awareness with recall is defined as 'both conscious experience and memory of events during surgery'. Perceptions of sound, pain or paralysis and assimilation of these in the memory can lead to post-traumatic stress disorder. Prospective studies report an incidence of 0.1-1%. Risk factors include those circumstances that result in under-dosing of anaesthetic agents relative to the patient's specific requirement. A lack of reliable monitoring of depth of anaesthesia is a result of our limited knowledge of the effect of anaesthetics on consciousness and memory. No additional benefit has been shown for the use of bispectral index (BIS) monitoring compared with measuring the expired concentration of an inhaled anaesthetic agent. High-risk patients undergoing total intravenous anaesthesia may benefit from BIS for monitoring depth of anaesthesia. Further scientific investigation of the neurological processes involved in awareness with recall is required in order to develop novel monitoring techniques.


Interleukin-15 augments superoxide production and microbicidal activity of human monocytes against Candida albicans.
Interleukin-15 (IL-15) is a newly described cytokine that shares biological activities with IL-2. We report here results demonstrating the ability of IL-15 to enhance superoxide production and antifungal activity of human monocytes. After 18 and 48 h of treatment with IL-15, human elutriated monocytes manifested enhanced superoxide production in response to either phorbol myristate acetate or opsonized Candida albicans blastoconidia. Similar results were obtained when monocytes were treated with IL-2, but to a lesser extent. Combination studies with IL-15 and IL-2 showed no additive or synergistic effects. Following incubation of monocytes with IL-15 for 18 h, there was no significant increase in mRNA transcripts for components of the NADPH oxidase complex, p40-phox, p47-phox, and gp91-phox, suggesting a posttranscriptional modulation of enhanced superoxide production. Antibodies against the gamma chain of the IL-2 receptor and, to a lesser extent, against the beta chain partially abrogated the IL-15-mediated enhanced superoxide production. Additionally, human monocytes showed enhanced killing activity against C. albicans after 18 h of incubation with IL-15 or IL-2, but this treatment did not enhance the ability of these cells to phagocytose the organism. In addition, the enhanced fungicidal activity seen after 18 h of treatment was no longer detectable after 48 h of cytokine treatment. Culture supernatants from the IL-15-treated monocytes were assayed for the presence of other proinflammatory cytokines. IL-15 treatment did not induce the release of detectable levels of tumor necrosis factor alpha, IL-1beta, or IL-12. Our results indicate that IL-15 upregulates the microbicidal activity of human monocytes against C. albicans.


Autonomy, choice, and decision making: how nursing home social workers view their role.
Open-ended interviews with 17 social workers from rural and urban Minnesota nursing homes explore how the practitioners perceive their roles in fostering the autonomy and decision making of residents. The social workers' espoused theory of practice is organized under three dimensions of autonomy: free action (supporting residents' choices), decision making (helping residents deliberate effectively), and continuity (maintaining a sense of self). The interviewees strongly identify with an advocacy role; helping residents make decisions appears less salient. The discussion explores the interviewees' perspectives and strategies, and suggests further research questions.


A retrospective study of pediatric renal trauma: A single-center experience in Japan.
To report pediatric renal trauma experiences at the National Center for Child Health and Development in Japan according to the Japanese Association for the Surgery of Trauma (JAST) classification 2008. Medical records were retrospectively reviewed for 45 children younger than 18 years old diagnosed with renal trauma from February 2004 to December 2021, regarding details of external causes, treatments, complications, and injury scales according to the JAST classification 2008. The cases who cannot be classified into the JAST classification 2008 will be categorized into our original type 0. There were 24 males and 21 females with a mean age of 8.5 years. Left kidneys were the predominantly affected side. Blunt injury was involved in every case (mainly falls and traffic accidents). Concomitant organ injuries were found in 13 cases. The injury scales were type Ia (13.3%), II (11.1%), IIIa (13.3%), IIIb (24.4%). Type 0 accounted for 37.8%, which were type 0c (congenital anomalies of the kidney and urinary tract without hematoma and/or laceration of kidney parenchyma) at 11.1% and type 0h (only hematuria and normal radiologic finding) at 26.7%. Treatments were blood transfusion, ureteral stenting, nephrostomy, and no nephrectomy. Complications were pseudoaneurysm, hypertension, and infection. Nearly 38% of cases cannot be classified into the JAST classification 2008, comprising at least two irrelevant types (type 0c and type 0h ). Accordingly, a pediatric renal injury scale should be established to achieve the precise diagnosis and treatments. However, further studies are still needed.


Heterogeneity in the spatial distribution of bacterial sexually transmitted infections.
Detailed knowledge of the spatial distribution of disease is required to inform service delivery and plan effective interventions. In order to elucidate the spatial epidemiology of three common sexually transmitted infections (STI) further, any significant spatiotemporal clustering of gonorrhoea, chlamydia or syphilis cases in New South Wales (NSW) was detected and described. Eleven years of notified STI case data were analysed. Calculation of age and sex-stratified incidence rates was followed by spatiotemporal cluster analyses to investigate differences in the epidemiology of gonorrhoea, chlamydia and syphilis. More than one-third of all gonorrhoea, chlamydia and syphilis cases in NSW were detected within cluster areas. Gonorrhoea cases were the most highly clustered, followed by syphilis, then chlamydia. Clusters were highly significant and relative risk estimates ranged from 1.6 to 22.9. The findings establish the high degree of geographical heterogeneity in STI incidence in NSW and indicate that the postal area of residence is an important predictor of STI incidence. Geographical surveillance could be incorporated into routine STI surveillance to identify populations in need of intervention. The evidence presented in this report indicates a need to implement geography-specific and phase-appropriate STI prevention and control strategies.


Fall and suffocation injuries associated with in-home use of car seats and baby carriers.
This paper examines two hazard patterns associated with in-home use and placement of car seats and baby carriers: 1) fall injuries resulting from placement on elevated surfaces and 2) suffocation resulting from seat overturn on soft surfaces. Emergency department data reporting on falls suffered by infants aged 6 months and younger, and associated with car seats and baby carriers, were provided by the U.S. Consumer Product Safety Commission's National Electronic Injury Surveillance System for 1997. National estimates were based on 396 incidents. Additionally, 15 cases of suffocation resulting from car/infant seat overturn on soft surfaces were identified through the Commission's Death Certificate and Investigation Files. In 1997, it is estimated that approximately 8700 infants were treated in emergency departments as a result of fall injuries suffered while using a car seat or baby carrier. More than one third of estimated falls were specifically attributed to the seat being placed on an elevated surface in the home, such as a counter or table. A search of suffocation incidents resulting from seat overturn on a soft surface produced 15 incidents. The majority of incidents occurred when the seat had been placed on a bed or waterbed. Many parents are unaware of hazards associated with common placement scenarios for infant car seats and baby carriers. These hazard patterns and their potential consequences need to be conveyed to new parents by pediatricians. At the same time, manufacturers have an obligation to pursue design alternatives that will reduce the likelihood of seat overturn.


RING domain dimerization is essential for RNF4 function.
RNF4 [RING (really interesting new gene) finger protein 4] family ubiquitin ligases are RING E3 ligases that regulate the homoeostasis of SUMOylated proteins by promoting their ubiquitylation. In the present paper we report that the RING domain of RNF4 forms a stable dimer, and that dimerization is required for ubiquitin transfer. Our results suggest that the stability of the E2~ubiquitin thioester bond is regulated by RING domain dimerization.


An ecological study of skin biopsies and skin cancer treatment procedures in the United States Medicare population, 2000 to 2015.
Analyses of skin cancer procedures adjusted for population changes are needed. To describe trends in skin cancer-related biopsies and procedures in Medicare beneficiaries. An ecological study of Medicare claims for skin biopsies and skin cancer procedures in 2000 to 2015. Biopsies increased 142%, and skin cancer procedures increased 56%. Mohs micrographic surgery (MMS) utilization increased on the head/neck, hands/feet, and genitalia (increasing from 11% to 27% of all treatment procedures) but was low on the trunk/extremities (increasing from 1% to 4%). Adjusted for increased Medicare enrollment (+36%) between 2000 and 2015, the number of biopsies and MMS procedures performed per 1000 beneficiaries increased (from 56 to 99 and from 5 to 15, respectively), whereas the number of excisions and destructions changed minimally (from 18 to 16 and from 19 to 18, respectively). Growth in biopsies and MMS procedures slowed between each time period studied: 4.3 additional biopsies per year and 0.9 additional MMS procedures per year per 1000 beneficiaries between 2000 and 2007, 2.2 and 0.5 more between 2008 and 2011, and 0.5 and 0.3 more between 2012 and 2015, respectively. Medicare claims-level data do not provide patient-level or nonsurgical treatment information. The increased number of skin cancer procedures performed was largely the result of Medicare population growth over time. MMS utilization increased primarily on high- and medium-risk and functionally and cosmetically significant locations where tissue sparing and maximizing cure are critical.


Expression of Ia antigens on myeloid progenitor cells in chronic myeloid leukemia: direct analysis using partially purified colony-forming cells.
The regulation of Ia (HLA-DR) antigen expression on myeloid progenitor cells may be closely related to the control of myelopoiesis in both normal individuals and chronic myeloid leukemia (CML) patients. In an effort to study directly the expression and behavior of Ia surface molecules on myeloid progenitor cells, we used an immunologic purification technique to enrich these cells approximately 100-fold from the peripheral blood of CML patients. The majority of cells in this blast population expressed HLA-DR antigens. Thirty percent to 40% of cells could form a granulocyte or monocyte colony in agar, and these cells tended to express the highest levels of HLA-DR. The number of HLA-DR molecules per cell increased about twofold as the cells tranversed the cell cycle from G0/G1 to G2/M. This was true for unstimulated cells or cells exposed to colony-stimulating factors. Some of this increase was related to a corresponding increase in cell size and is also seen with other cell surface antigens such as beta-2-microglobulin. Ia antigen expression was not modified by culture with colony-stimulating factors, fetal calf serum, or serum-free, prostaglandin-free medium for periods of up to 24 hours. These results demonstrate that Ia antigens are expressed on the myeloid progenitor cells of CML, are increased in the S and G2/M phases of the cell cycle, and are stable under most in vitro culture conditions for at least 24 hours of culture.


Gene expression profiling of vasoregression in the retina--involvement of microglial cells.
Vasoregression is a hallmark of vascular eye diseases but the mechanisms involved are still largely unknown. We have recently characterized a rat ciliopathy model which develops primary photoreceptor degeneration and secondary vasoregression. To improve the understanding of secondary vasoregression in retinal neurodegeneration, we used microarray techniques to compare gene expression profiles in this new model before and after retinal vasoregression. Differential gene expression was validated by quantitative RT-PCR, Western blot and immunofluorescence. Of the 157 genes regulated more than twofold, the MHC class II invariant chain CD74 yielded the strongest upregulation, and was allocated to activated microglial cells close to the vessels undergoing vasoregression. Pathway clustering identified genes of the immune system including inflammatory signaling, and components of the complement cascade upregulated during vasoregression. Together, our data suggest that microglial cells involved in retinal immune response participate in the initiation of vasoregression in the retina.


Arabidopsis thaliana GPAT8 and GPAT9 are localized to the ER and possess distinct ER retrieval signals: functional divergence of the dilysine ER retrieval motif in plant cells.
Glycerol-3-phosphate acyltransferase (GPAT; EC 188.8.131.52) catalyzes the committed step in the production of glycerolipids, which are major components of cellular membranes, seed storage oils, and epicuticular wax coatings. While the biochemical activities of GPATs have been characterized in detail, the cellular features of these enzymes are only beginning to emerge. Here we characterized the phylogenetic relationships and cellular properties of two GPAT enzymes from the relatively large Arabidopsis thaliana GPAT family, including GPAT8, which is involved in cutin biosynthesis, and GPAT9, which is a new putative GPAT that has extensive homology with a GPAT from mammalian cells involved in storage oil formation and, thus, may have a similar role in plants. Immunofluorescence microscopy of transiently-expressed myc-epitope-tagged GPAT8 and GPAT9 revealed that both proteins were localized to the endoplasmic reticulum (ER), and differential permeabilization experiments indicated that their N- and C-termini were oriented towards the cytosol. However, these two proteins contained distinct types of ER retrieval signals, with GPAT8 possessing a divergent type of dilysine motif (-KK-COOH rather than the prototypic -KKXX-COOH or -KXKXX-COOH motif) and GPAT9 possessing a hydrophobic pentapeptide motif (-phi-X-X-K/R/D/E-phi-; where phi are large hydrophobic amino acid residues). Notably, the divergent dilysine motif in GPAT8 only functioned effectively when additional upstream residues were included to provide the proper protein context. Extensive mutational analyses of the divergent dilysine motif, based upon sequences present in the C-termini of other GPAT8s from various plant species, further expanded the functional definition of this molecular targeting signal, thereby providing insight to the targeting signals in other GPAT family members as well as other ER-resident membrane proteins within plant cells.


Colocalization of synaptophysin with transferrin receptors: implications for synaptic vesicle biogenesis.
We have reported previously that the synaptic vesicle (SV) protein synaptophysin, when expressed in fibroblastic CHO cells, accumulates in a population of recycling microvesicles. Based on preliminary immunofluorescence observations, we had suggested that synaptophysin is targeted to the preexisting population of microvesicles that recycle transferrin (Johnston, P. A., P. L. Cameron, H. Stukenbrok, R. Jahn, P. De Camilli, and T. C. Südhof. 1989. EMBO (Eur. Mol. Biol. Organ.) J. 8:2863-2872). In contrast to our results, another group reported that expression of synaptophysin in cells which normally do not express SV proteins results in the generation of a novel population of microvesicles (Leube, R. E., B. Wiedenmann, and W. W. Franke. 1989. Cell. 59:433-446). We report here a series of morphological and biochemical studies conclusively demonstrating that synaptophysin and transferrin receptors are indeed colocalized on the same vesicles in transfected CHO cells. These observations prompted us to investigate whether an overlap between the distribution of the two proteins also occurs in endocrine cell lines that endogenously express synaptophysin and other SV proteins. We have found that endocrine cell lines contain two pools of membranes positive for synaptophysin and other SV proteins. One of the two pools also contains transferrin receptors and migrates faster during velocity centrifugation. The other pool is devoid of transferrin receptors and corresponds to vesicles with the same sedimentation characteristics as SVs. These findings suggest that in transfected CHO cells and in endocrine cell lines, synaptophysin follows the same endocytic pathway as transferrin receptors but that in endocrine cells, at some point along this pathway, synaptophysin is sorted away from the recycling receptors into a specialized vesicle population. Finally, using immunofluorescent analyses, we found an overlap between the distribution of synaptophysin and transferrin receptors in the dendrites of hippocampal neurons in primary cultures before synapse formation. Axons were enriched in synaptophysin immunoreactivity but did not contain detectable levels of transferrin receptor immunoreactivity. These results suggest that SVs may have evolved from, as well as coexist with, a constitutively recycling vesicular organelle found in all cells.


Pathogen-specific risk of celiac disease following bacterial causes of foodborne illness: a retrospective cohort study.
The US CDC recently estimated over 2 million foodborne illnesses annually are caused by 4 major enteropathogens: non-typhoid Salmonella spp., Campylobacter spp., Shigella spp., and Yersinia enterocolitica. While recent data suggest functional gastrointestinal disorders are associated with these infections, studies linking foodborne illness to celiac disease (CD) are limited. We utilized a US Department of Defense medical encounter database to evaluate the risk of CD following select foodborne infections. We identified subjects with acute gastroenteritis between 1998 and 2009 attributed to Salmonella (nontyphoidal) spp., Shigella spp., Campylobacter spp., or Y. enterocolitica and matched each with up to 4 unexposed subjects. Exposed and unexposed subjects were followed for incident CD diagnosis for their entire military record duration (or a minimum of 1 year). Relative risks were calculated using modified Poisson regression to determine the relationship between pathogen-attributable gastroenteritis and CD while controlling for covariates. A total of 1,753 pathogen-specific gastroenteritis cases (Campylobacter: 738; Salmonella: 624; Shigella: 376; Yersinia: 17) were identified and followed for a median of 3.8 years. The incidence (per 100,000 person-years) of CD was 0.05. We found a suggested risk of CD after Campylobacter, but not other foodborne infection etiologies. These data support a previous study demonstrating increased risk of CD following Campylobacteriosis and highlight the need for additional research into how infections might trigger CD in susceptible individuals.


Multiplanar metrizamide-enhanced CT imaging of the foramen magnum.
Definition of masses and malformations at the foramen magnum has been less than optimal with traditional radiographic techniques. The use of intrathecal metrizamide with computed tomographic (CT) scanning improves contrast resolution and facilitates detection of abnormalities that may not be apparent on Pantopaque cisternography, plain films, or conventional axial CT alone. Fifty patients with clinically suspected foramen-magnum lesions were evaluated with this technique. Forty were abnormal with the following findings: 14 cases of the Chiari malformation (five cases had associated syringomyelia), five meningiomas, three neuromas, a chondrosarcoma, a chordoma, and five cervical and three brainstem gliomas. Six cases of syringomyelia, a large posterior-fossa subarachnoid cyst, and an anomaly of the cisterna magna were also diagnosed. Sagittal and coronal images were beneficial in defining tonsillar position, configuration of the fourth ventricle, and the relations of mass lesions to the spinal canal and subarachnoid space. Metrizamide CT cisternography provides superior spatial and contrast resolution of lesions at the foramen magnum not obtainable with other radiographic techniques. Morbidity is minimal because of the low dose of metrizamide used. Multiplanar reformations are particularly helpful in assessing anatomic detail not readily recognized on axial scans alone.


Distinct latent profiles based on neurobehavioural, physical and psychosocial functioning of former National Football League (NFL) players: an NFL-LONG Study.
To identify subgroups of former National Football League (NFL) players using latent profile analysis (LPA) and examine their associations with total years of participation (TYP) and self-reported lifetime sport-related concussion history (SR-CHx). Former NFL players (N=686) aged 50-70 years, with an average 18.0 TYP (±4.5) completed a questionnaire. SR-CHx distributions included: low (0-3; n=221); intermediate (4-8; n=209) and high (9+; n=256). LPA measures included: Quality of Life in Neurological Disorders Emotional-Behavioral Dyscontrol, Patient Reported Outcomes Measurement Information System Cognitive Function, Emotional Support, Self-Efficacy, Meaning and Purpose, Physical Function, Pain Interference, Participation in Social Roles and Activities, Anxiety, Depression, Fatigue, and Sleep Disturbance. Demographic, medical/psychiatric history, current psychosocial stressors, TYP and SR-CHx were compared across latent profiles (LPs). A five profile solution emerged: (LP1) global higher functioning (GHF; 26.5%); (LP2) average functioning (10.2%); (LP3) mild somatic (pain and physical functioning) concerns (22.0%); (LP4) somatic and cognitive difficulties with mild anxiety (SCA; 27.5%); LP5) global impaired functioning (GIF; 13.8%). The GIF and SCA groups reported the largest number ofe- medical/psychiatric conditions and higher psychosocial stressor levels. SR-CHx was associated with profile group (χ2(8)=100.38, p<0.001); with a higher proportion of GIF (72.6%) and SCA (43.1%) groups reporting being in the high SR-CHx category, compared with GHF (23.1%), average (31.4%) and somatic (27.8%) groups. TYP was not significantly associated with group (p=0.06), with greater TYP reported by the GHF group. Five distinct profiles of self-reported functioning were identified among former NFL players. Several comorbid factors (ie, medical/psychiatric diagnoses and psychosocial stressors) and SR-CHx were associated with greater neurobehavioural and psychosocial dysfunction.