Investigation of the force associated with the formation of lacerations and skull fractures.
Post-mortem examination is often relied upon in order to determine whether a suspicious death was natural, accidental, suicidal or homicidal. However, in many cases the mechanism by which a single injury has been inflicted cannot be determined with certainty based on pathological examination alone. Furthermore the current method of assessing applied force relating to injury is restricted to an arbitrary and subjective scale (mild, moderate, considerable, or severe). This study investigates the pathophysiological nature of head injuries caused by blunt force trauma, specifically in relation to the incidence and formation of a laceration. An experimental model was devised to assess the force required to cause damage to the scalp and underlying skull of porcine specimens following a single fronto-parietal impact. This was achieved using a drop tower equipped with adapted instrumentation for data acquisition. The applied force and implement used could be correlated with resultant injuries and as such aid pathological investigation in the differentiation between falls and blows. Experimentation revealed prevalent patterns of injury specific to the reconstructed mechanism involved. It was found that the minimum force for the occurrence of a laceration was 4,000 N.


Validity of diagnosis and procedure codes for identifying neural tube defects in infants.
The use of validated criteria to identify birth defects in electronic healthcare databases can avoid the cost and time-intensive efforts required to conduct chart reviews to confirm outcomes. This study evaluated the validity of various case-finding methodologies to identify neural tube defects (NTDs) in infants using an electronic healthcare database. This analysis used data generated from a study whose primary aim was to evaluate the association between first-trimester maternal prescription opioid use and NTDs. The study was conducted within the Medication Exposure in Pregnancy Risk Evaluation Program. A broad approach was used to identify potential NTDs including diagnosis and procedure codes from inpatient and outpatient settings, death certificates and birth defect flags in birth certificates. Potential NTD cases were chart abstracted and confirmed by clinical experts. Positive predictive values (PPVs) and 95% confidence intervals (95% CI) are reported. The cohort included 113 168 singleton live-born infants: 55 960 infants with opioid exposure in pregnancy and 57 208 infants unexposed in pregnancy. Seventy-three potential NTD cases were available for the validation analysis. The overall PPV was 41% using all diagnosis and procedure codes plus birth certificates. Restricting approaches to codes recorded in the infants' medical record or to birth certificate flags increased the PPVs (72% and 80%, respectively) but missed a substantial proportion of confirmed NTDs. Codes in electronic healthcare data did not accurately identify confirmed NTDs. These results indicate that chart review with adjudication of outcomes is important when conducting observational studies of NTDs using electronic healthcare data.


Preparation, optimisation and characterisation of lyophilised rapid disintegrating tablets based on gelatin and saccharide.
Despite recent success, rapidly disintegrating lyophilized tablets still face problems of low mechanical strength and higher disintegration times resulting in poor patient compliance. The aim of the current work was to carry out a systematic study to understand the factors controlling mechanical properties of these formulations. The work investigated the influence of two bloom strengths of gelatin: low (60 bloom) and high (225 bloom) at different stock solution concentrations (2, 5, 7.5, and 10 %w/w). This was followed by investigation of addition of five saccharides (xylitol, glucose, trehalose, maltotriose and mannitol) at varied concentration range (10-80 %w/w) to decipher their influence on disintegration time, mechanical and thermal properties of the formulation. The results indicated that the disintegration time of the tablets dramatically decreased by decreasing the concentration and bloom strength of gelatin in the stock solution. However the mechanical properties of the tablets were mainly influenced by the concentration of gelatin rather than the bloom strength. The addition of saccharides resulted in enhancement of tablet properties and was concentration dependent. All the saccharides improved the fractubility of the tablets significantly at high concentration (equal or higher than 40% w/w). However, only high concentration (equal or higher than 40% w/w) of trehalose, maltotriose and mannitol significantly enhanced the hardness. Additionally, mannitol crytallised during freeze drying and consequently produced elegant tablets, whilst the other saccarides exhibited lyoprotectant activity as they were able to retain amorphous status. Based on the above findings, an optimized formulation was also successfully developed and characterized to deliver 100 microg dose of Clonidine HCl.


Drivers of the dive response in trained harbour porpoises (Phocoena phocoena).
Pronounced dive responses through peripheral vasoconstriction and bradycardia enable prolonged apnoea in marine mammals. For most vertebrates, the dive response is initiated upon face immersion, but little is known about the physical drivers of diving and surfacing heart rate in cetaceans whose faces are always mostly submerged. Using two trained harbour porpoises instrumented with an ECG-measuring sound-and-movement tag (DTAG-3), we investigated the initiation and progression of bradycardia and tachycardia during apnoea and eupnoea for varying levels of immersion. We show that paranasal wetting drives bradycardia initiation and progression, whereas apnoea leads to dive-level bradycardia eventually, but not instantly. At the end of dives, heart rate accelerates independently of lung expansion, perhaps in anticipation of surfacing; however, full tachycardia is only engaged upon inhalation. We conclude that breathing drives surface tachycardia, whereas blowhole wetting is an important driver of bradycardia; however, anticipatory/volitional modulation can overrule such responses to sensory inputs.


Anti-HLA-DR/anti-DOTA diabody construction in a modular gene design platform: bispecific antibodies for pretargeted radioimmunotherapy.
Recombinant immunoglobulin libraries of single chain molecules (sc) from the variable domains of antibody light and heavy chains (Fv), have great promise for new approaches to radioimmunotherapy (RIT). However, creating and evaluating scFv from diverse sources is time consuming and differences in molecular format can influence in vitro and in vivo characteristics. Furthermore, scFv do not have optimal characteristics for targeting therapy to tumor because of their small size and univalent binding. Diabody molecules at least twice the size of scFv are better for RIT because bivalent and bispecific molecules can be constructed. A polymerase chain reaction (PCR) based primer system was created to easily convert scFv genes into a diabody gene format, once they have been placed into pCANTAB 5E, a readily available vector. The primer system for this diabody gene platform was developed and tested by constructing an anti-lymphoma/anti-chelate, bispecific diabody (anti-HLA-DR/anti-DOTA). Two mouse scFv libraries were screened for reactive clones using recombinant phage display techniques. Selected mouse anti-HLA-DR and anti-DOTA scFv genes were combined, ligated into the pCANTAB 5E vector that co-expressed these self-assembling scFv in E. coli as two mismatched nonlinked pairs (VHA-link-VLB; VHB-link-VLA). The diabody protein that was purified from periplasm had the expected molecular characteristics when analyzed by sequencing, chromatography, electrophoresis and Western blot. This modular gene design platform provides methodology for easy and rapid creation of diabody molecules from diverse scFv libraries. Diabodies from various scFv can easily be produced, thereby facilitating comparative preclinical studies en route to development of new tumor targeting molecules.


Targeted Analysis of Serum Proteins Encoded at Known Inflammatory Bowel Disease Risk Loci.
Few studies have investigated the blood proteome of inflammatory bowel disease (IBD). We characterized the serum abundance of proteins encoded at 163 known IBD risk loci and tested these proteins for their biomarker discovery potential. Based on the Human Protein Atlas (HPA) antibody availability, 218 proteins from genes mapping at 163 IBD risk loci were selected. Targeted serum protein profiles from 49 Crohn's disease (CD) patients, 51 ulcerative colitis (UC) patients, and 50 sex- and age-matched healthy individuals were obtained using multiplexed antibody suspension bead array assays. Differences in relative serum abundance levels between disease groups and controls were examined. Replication was attempted for CD-UC comparisons (including disease subtypes) by including 64 additional patients (33 CD and 31 UC). Antibodies targeting a potentially novel risk protein were validated by paired antibodies, Western blot, immuno-capture mass spectrometry, and epitope mapping. By univariate analysis, 13 proteins mostly related to neutrophil, T-cell, and B-cell activation and function were differentially expressed in IBD patients vs healthy controls, 3 in CD patients vs healthy controls and 2 in UC patients vs healthy controls (q < 0.01). Multivariate analyses further differentiated disease groups from healthy controls and CD subtypes from UC (P < 0.05). Extended characterization of an antibody targeting a novel, discriminative serum marker, the laccase (multicopper oxidoreductase) domain containing 1 (LACC1) protein, provided evidence for antibody on-target specificity. Using affinity proteomics, we identified a set of IBD-associated serum proteins encoded at IBD risk loci. These candidate proteins hold the potential to be exploited as diagnostic biomarkers of IBD.


Occupational transmission of Mycobacterium tuberculosis to health care workers in a university hospital in Lima, Peru.
From November 1996 through March 1997, presumptive active pulmonary tuberculosis (TB) was detected in 44 health care workers (HCWs) at a university hospital in Lima, Peru. To further assess the magnitude of the outbreak and determine risk factors for occupational Mycobacterium tuberculosis transmission, we identified HCWs in whom active pulmonary TB was diagnosed from January 1994 through January 1998, calculated rates by year and hospital work area, and conducted a tuberculin skin test (TST) survey. Thirty-six HCWs had confirmed active pulmonary TB. The rate of TB was significantly higher among the 171 HCWs employed in the laboratory than among HCWs employed in all other areas. In multivariate analysis, the only independent risk factor for HCW M. tuberculosis infection in HWCs employed in the laboratory was the use of common staff areas. Very high rates of active pulmonary TB were detected among HCWs at this hospital, and occupational acquisition in the laboratory was associated with HCW-to-HCW transmission.


From childhood trauma to hyperarousal in adults: The mediating effect of maladaptive shame coping and insomnia.
A new line of insomnia research focuses on the developmental trajectories from early live stress to insomnia in adulthood. Adverse childhood experiences (ACE's) might create a vulnerability for later maladaptive coping with distress, as seen in chronic hyperarousal or insomnia. In an functional magnetic resonance imaging (fMRI) study, failure to dissociate the neurobiological components of shame from autobiographical shameful memories in insomnia was reflected by continued activation of the dorsal anterior cingulate cortex (dACC), which may be a result of maladaptive coping in the wake of ACE's. Following up on that study, the current pilot study explores the relation between ACE's, shame coping-styles, adult insomnia, hyperarousal, and neurobiology of autobiographical memory. We used existing data (N = 57) from individuals with insomnia (N = 27) and controls (N = 30), and asked these participants to complete the childhood trauma questionnaire (CTQ). Two structural equation models were used to test the hypotheses that shame-coping styles and insomnia symptom severity mediate the association between ACE's and (1) self-rated hyperarousal symptoms and (2) dACC activation to recall of autobiographical memories. For the association between ACE's and hyperarousal, there was a significant mediation of shame-coping style (p < 0.05). This model also indicated worse shame coping with more ACE's (p < 0.05) and worse insomnia symptoms with more ACES's (p < 0.05), but no association between shame coping and insomnia symptoms (p = 0.154). In contrast, dACC activation to recall of autobiographical memories could only be explained by its direct association with ACE's (p < 0.05), albeit that in this model more ACE's were also associated with worse insomnia symptoms. These findings could have an implication for the approach of treatment for insomnia. It could be focused more on trauma and emotional processing instead of conventional sleep interventions. Future studies are recommended to investigate the relationship mechanism between childhood trauma and insomnia, with additional factors of attachment styles, personality, and temperament.


The effects of pollution on fish health.
Potentially harmful substances-e.g. pesticides, heavy metals and hydrocarbons-are often released into the aquatic environment. When large quantities of pollutants are released there may be an immediate impact as measured by large-scale sudden mortalities of aquatic organisms, e.g. fish kills resulting from contamination of waterways with agricultural pesticides. Lower levels of discharge may result in an accumulation of the pollutants in aquatic organisms. The end results, which may occur long after the pollutants have passed through the environment, include immunosuppression, reduced metabolism, and damage to gills and epithelia. However, the link between adverse water quality and fish diseases is not proven. Alleged pollution-related diseases include epidermal papilloma, fin/tail rot, gill disease, hyperplasia, liver damage, neoplasia and ulceration. Many surveys have indicated a greater proportion of diseased fish in polluted compared to non-polluted marine sites. Yet, the value of such surveys may be questioned. Specific examples of fish diseases thought to reflect the effects of pollution include surface lesions attributed to Serratia plymuthica, fin and tail rot caused by Aeromonas hydrophila and Pseudomonas fluorescens, gill disease resulting from the actvity of Flavobacterium spp., vibriosis as caused by Vibrio anguillarum, and enteric redmouth (causal agent, Yersinia ruckeri). Research indicated that some of the diseases caused by Aeromonas, Flavobacterium and Pseudomonas resulted from generally adverse water quality, i.e. higher than usual quantities of organic material, oxygen depletion, changes in pH values and enhanced microbial populations. Some infections with Serratia and Yersina may well have reflected contamination of waterways with domestic sewage, e.g. leaking septic tanks. At least one outbreak of vibriosis was linked to high concentrations of copper, which may have debilitated the fish making them more susceptible to disease.


Eye findings in 8 children and a spontaneously aborted fetus with RSH/Smith-Lemli-Opitz syndrome.
We evaluate the ophthalmologic findings in 8 children with RSH/Smith-Lemli-Opitz syndrome (SLOS) and document abnormal concentrations of cholesterol and cholesterol precursors in the ocular tissues in a case of SLOS. The most common ophthalmologic finding was blepharoptosis, which was found in 6 of 8 patients, with the severity ranging from mild to moderate. None of the patients in the present study demonstrated cataracts; none had amblyopia from blepharoptosis. One patient had a right hypertropia with overaction of the inferior oblique muscle. This patient also had optic atrophy and a second patient had bilateral optic nerve hypoplasia. The importance of these findings to the visual function remains to be defined. Sterol analysis from ocular tissues of an aborted fetus with SLOS showed increased 7- and 8-dehydrocholesterol and a low cholesterol concentration in the retinal pigment epithelium, lens, cornea, and sclera. Routine ophthalmologic examination is indicated in SLOS because of the high incidence of abnormalities, most likely due to the abnormal synthesis of cholesterol and cholesterol precursors in the ocular tissues of these patients, as evidenced by sterol analysis of the ocular tissues in a case of SLOS.


Functional Microarchitecture of the Mouse Dorsal Inferior Colliculus Revealed through In Vivo Two-Photon Calcium Imaging.
The inferior colliculus (IC) is an obligatory relay for ascending auditory inputs from the brainstem and receives descending input from the auditory cortex. The IC comprises a central nucleus (CNIC), surrounded by several shell regions, but the internal organization of this midbrain nucleus remains incompletely understood. We used two-photon calcium imaging to study the functional microarchitecture of both neurons in the mouse dorsal IC and corticocollicular axons that terminate there. In contrast to previous electrophysiological studies, our approach revealed a clear functional distinction between the CNIC and the dorsal cortex of the IC (DCIC), suggesting that the mouse midbrain is more similar to that of other mammals than previously thought. We found that the DCIC comprises a thin sheet of neurons, sometimes extending barely 100 μm below the pial surface. The sound frequency representation in the DCIC approximated the mouse's full hearing range, whereas dorsal CNIC neurons almost exclusively preferred low frequencies. The response properties of neurons in these two regions were otherwise surprisingly similar, and the frequency tuning of DCIC neurons was only slightly broader than that of CNIC neurons. In several animals, frequency gradients were observed in the DCIC, and a comparable tonotopic arrangement was observed across the boutons of the corticocollicular axons, which form a dense mesh beneath the dorsal surface of the IC. Nevertheless, acoustically responsive corticocollicular boutons were sparse, produced unreliable responses, and were more broadly tuned than DCIC neurons, suggesting that they have a largely modulatory rather than driving influence on auditory midbrain neurons. Due to its genetic tractability, the mouse is fast becoming the most popular animal model for sensory neuroscience. Nevertheless, many aspects of its neural architecture are still poorly understood. Here, we image the dorsal auditory midbrain and its inputs from the cortex, revealing a hitherto hidden level of organization and paving the way for the direct observation of corticocollicular interactions. We show that a precise functional organization exists in the mouse auditory midbrain, which has been missed by previous, more macroscopic approaches. The fine-scale distribution of sound-frequency tuning suggests that the mouse midbrain is more similar to that of other mammals than previously thought and contrasts with the more heterogeneous organization reported in imaging studies of auditory cortex.


Antioxidants improve early survival of cardiomyoblasts after transplantation to the myocardium.
We tested the hypothesis that modulation of the microenvironment (using antioxidants) will increase stem cell survival in hypoxia and after transplantation to the myocardium. Rat cardiomyoblasts were stably transfected with a reporter gene (firefly luciferase) for bioluminescence imaging (BLI). First, we examined the role of oxidative stress in cells under hypoxic conditions. Subsequently, stem cells were transplanted to the myocardium of rats using high-resolution ultrasound, and their survival was monitored daily using BLI. Under hypoxia, oxidative stress was increased together with decreased cell survival compared to control cells, both of which were preserved by antioxidants. In living subjects, oxidative stress blockade increased early cell survival after transplantation to the myocardium, compared to untreated cells/animals. Modulation of the local microenvironment (with antioxidants) improves stem cell survival. Increased understanding of the interaction between stem cells and their microenvironment will be critical to advance the field of regenerative medicine.


Measurement reproducibility in the early stages of biomarker development.
Biomarker discovery and development requires measurement reproducibility studies in addition to case-control studies. Parallel pursuit of reproducibility studies is especially important for emerging technologies such as protein biomarkers based on time-of-flight mass spectrometry, the case considered in this paper. For parallel studies, a way to improve reproducibility prior to identification of protein species is necessary. One approach is use of functional principal components analysis (PCA) as the basis for assessing measurement reproducibility. Reproducibility studies involve repeated measurement of a reference material such as a human serum standard. Measurement in our example is by SELDI-TOF (surface-enhanced laser desorption and ionization time-of-flight) mass spectrometry. Reproducibility is defined in reference to a source of variation, which in our example is associated with a type of commercially available protein biochip. We obtained spectra for 8 spots on each 11 chips. Two spectra are generally more alike when obtained from the same chip rather than different chips. Thus, our experiment indicates potential improvements from reducing variation in chip manufacture and chip handling during measurement. Our analysis involves careful registration of the spectra and characterization of the spectral differences. As shown by our example, a metrological analysis may enhance case-control studies by guiding optimization of the measurements underlying the biomarker.


A first step towards transparency in pricing of medicines and scheduled substances - publication of guidelines for pharmaco-economic submissions.
The National Department of Health of South Africa recently published guidelines for pharmaco-economic (PE) submissions in accordance with the Medicines and Related Substances Act (Act 101 of 1965), which came into effect on 1 April 2013. These guidelines relate to the compilation of PE submissions for evidence of cost-effectiveness of medicine or scheduled substances. The PE guidelines are a first step towards the creation of a mechanism whereby the value of medicine can be quantified in a transparent manner. The 'voluntary' nature of PE submissions speaks to the current lack of knowledge, understanding and capacity related to pharmaco-economics that exists in the private healthcare market. The current disconnect between the PE guidelines and the Medical Schemes Act should be addressed as a matter of urgency to provide a mechanism whereby guidance in terms of cost-effectiveness from a PE evaluation will be supported by guaranteed reimbursement by medical schemes.


Elastase-induced intracranial dolichoectasia model in mice.
Intracranial dolichoectasia is associated with high morbidity, and its pathophysiology remains poorly understood. To develop a technique for the creation of a murine model of dolichoectasia. C57/BL6 mice were injected with 0 milliunit (mu) (control, n = 5), 15 mu (n = 7), 25 mu (n = 10), 35 mu (n = 10), and 55 mu (n = 6) of elastase in the cisterna magna. Fourteen days after injection, the vasculature of the brain was perfused with MicroFil polymerizing compound. Tortuosity index and the percentage increase in arterial diameter were calculated for the basilar artery, posterior communicating arteries, and the A1 segment of the anterior cerebral arteries. Tortuosity index >10 combined with 25% increase in diameter were used to indicate success in achieving dolichoectasia. The mortality rate was 28%, 30%, 80%, and 83% in the 15, 25, 35, and 55 mu groups, respectively. As the 35 and 55 mu groups experienced unacceptable mortality rates, they were excluded from further analysis. The tortuosity index and percent increase arterial diameter of the 15 and 25 mu groups for the left anterior cerebral arteries, right anterior cerebral arteries, left posterior communicating arteries, right posterior communicating arteries, and basilar artery were significantly higher (TI >10 and arterial diameter >25%) than in the control. There was no significant difference in tortuosity index or artery diameter between the 15 and 25 mu groups for any of the 5 artery segments. Elastase injection through the cisterna magna can induce intracranial dolichoectasia in mice. Fifteen to 25 mu of elastase is an appropriate dose to use with acceptable mortality.


ACSM Expert Consensus Statement on Exertional Heat Illness: Recognition, Management, and Return to Activity.
Exertional heat stroke (EHS) is a true medical emergency with potential for organ injury and death. This consensus statement emphasizes that optimal exertional heat illness management is promoted by a synchronized chain of survival that promotes rapid recognition and management, as well as communication between care teams. Health care providers should be confident in the definitions, etiologies, and nuances of exertional heat exhaustion, exertional heat injury, and EHS. Identifying the athlete with suspected EHS early in the course, stopping activity (body heat generation), and providing rapid total body cooling are essential for survival, and like any critical life-threatening situation (cardiac arrest, brain stroke, sepsis), time is tissue. Recovery from EHS is variable, and outcomes are likely related to the duration of severe hyperthermia. Most exertional heat illnesses can be prevented with the recognition and modification of well-described risk factors ideally addressed through leadership, policy, and on-site health care.


DNA damage-binding complex recruits HDAC1 to repress Bcl-2 transcription in human ovarian cancer cells.
Elevated expression of the antiapoptotic factor Bcl-2 is believed to be one of the contributing factors to an increased relapse rate associated with multiple cisplatin-resistant cancers. DNA damage-binding protein complex subunit 2 (DDB2) has recently been revealed to play an important role in sensitizing human ovarian cancer cells to cisplatin-induced apoptosis through the downregulation of Bcl-2, but the underlying molecular mechanism remains poorly defined. Here, it is report that DDB2 functions as a transcriptional repressor for Bcl-2 in combination with DDB1. Quantitative ChIP and EMSA analysis revealed that DDB2 binds to a specific cis-acting element at the 5'-end of Bcl-2 P1 promoter. Overexpression of DDB2 resulted in marked losses of histone H3K9,14 acetylation along the Bcl-2 promoter and enhancer regions, concomitant with a local enrichment of HDAC1 to the Bcl-2 P1 core promoter in ovarian cancer cells. Coimmunoprecipitation and in vitro binding analyses identified a physical interaction between DDB1 and HDAC1, whereas downregulation of HDAC1 significantly enhanced Bcl-2 promoter activity. Finally, in comparison with wild-type DDB2, mutated DDB2, which is unable to repress Bcl-2 transcription, mediates a compromised apoptosis upon cisplatin treatment. Taken together, these data support a model wherein DDB1 and DDB2 cooperate to repress Bcl-2 transcription. DDB2 recognizes and binds to the Bcl-2 P1 promoter, and HDAC1 is recruited through the DDB1 subunit associated with DDB2 to deacetylate histone H3K9,14 across Bcl-2 regulatory regions, resulting in suppressed Bcl-2 transcription. Increasing the expression of DDB complex may provide a molecular strategy for cancer therapy.


The oncogene product Vav is a crucial regulator of primary cytotoxic T cell responses but has no apparent role in CD28-mediated co-stimulation.
The guanine nucleotide-exchange factor Vav is a regulator of antigen-mediated cytoskeletal reorganization required for receptor clustering, proliferation and thymic selection. Moreover, Vav has been identified as a major substrate in the CD28 signal transduction pathway and overexpression of Vav enhances TCR-mediated IL-2 secretion in T cells. Here we show that CD3- plus CD28-mediated proliferation and IL-2 production were reduced in vav gene-deficient T cells. However, Vav had no apparent role in phorbol 12-myristate 13-acetate-plus CD28-mediated proliferation and IL-2 production, suggesting that Vav acts downstream of the TCR/CD3 complex. In vivo, Vav expression was crucial to generate primary vesicular stomatitis virus (VSV)-specific cytotoxic T cell responses. In contrast, vav-/- mice exhibited a reduced but significant footpad swelling after lymphocytic choriomeningitis virus (LCMV) infections and mounted a measurable primary cytotoxic T cell response to LCMV. Upon in vitro restimulation, cytotoxic T cell responses of both VSV- and LCMV-infected mice reached near normal levels. Our data provide the first genetic evidence that Vav is an important effector molecule that relays antigen receptor signaling to IL-2 production and activation of cytotoxic T cells.


Obesity Epidemic: Is Its Impact on Total Joint Arthroplasty Underestimated? An Analysis of National Trends.
Obesity is a well-established risk factor for total joint arthroplasty (TJA) and a number of complications including prosthetic joint infection. The annual changes in the prevalence of obesity among primary, revision, and infected TJA has not been studied at a national level. Given the higher costs of complications of TJA, it is important to understand the association of obesity with the annual trends of revision and infected TJA. (1) Is the prevalence of obesity increasing among patients undergoing THA/TKA? (2) Is the prevalence of obesity increasing among patients undergoing revision THA/TKA? (3) Is the prevalence of obesity increasing among patients with infected THA/TKA? Annual volumes of primary, revision, and infected THA and TKA from 1998 to 2011 were obtained from the Nationwide Inpatient Sample. Using mathematical equations, the prevalence of obesity was estimated from relative risks and national obesity prevalence. National obesity prevalence was obtained from public health sources and the relative risk estimates were obtained from previously published meta-analyses and population-based studies. Annual prevalence of obesity was obtained by dividing the number of obese primary/revision/infected procedures in each year by the total number of corresponding procedures in that year. Annual changes in the prevalence of obesity were analyzed using linear regression. The prevalence of obesity is increasing among patients undergoing THA (1998: 60,264 of 154,337 [39%], 2011: 160,241 of 305,755 [52%], increase of 1.05%/year [confidence interval {CI}, 0.95%-1.15%], p < 0.001) and TKA (1998: 143,681 of 251,309 [57%], 2011: 448,712 of 644,243 [70%], increase of 0.97%/year [CI, 0.87%-1.07%], p < 0.001). There was an increasing prevalence of obesity with THA revisions (1998: 16,322 of 34,139 [48%], 2011: 33,304 of 54,453 [61%], increase of 1.04%/year [CI, 0.94%-1.15%], p < 0.001) and in TKA revisions (1998: 16,837 of 26,539 [63%], 2011: 52,151 of 69,632 [75%], increase of 0.89%/year [CI, 0.79%-0.99%], p < 0.001). There was an increasing prevalence of obesity with THA infections (1998: 2068 of 3018 [69%], 2011: 6856 of 8687 [79%], increase of 0.80%/year [CI, 0.71%-0.89%], p < 0.001) and in TKA infections (1998: 3563 of 4684 [76%], 2011: 14,178 of 16,774 [85%], increase of 0.65%/year [CI, 0.57%-0.73%], p < 0.001). The prevalence of obesity has increased in patients undergoing primary, revision, and infected TJA in United States. The obesity epidemic appears to be related to the growing trends of revision and infection after TJA. With the obesity rates expected to grow further, the revision and infection burden associated with obesity may increase in the future. Level II, prognostic study.


Effects of HCM cTnI mutation R145G on troponin structure and modulation by PKA phosphorylation elucidated by molecular dynamics simulations.
Cardiac troponin (cTn) is a key molecule in the regulation of human cardiac muscle contraction. The N-terminal cardiac-specific peptide of the inhibitory subunit of troponin, cTnI (cTnI(1-39)), is a target for phosphorylation by protein kinase A (PKA) during β-adrenergic stimulation. We recently presented evidence indicating that this peptide interacts with the inhibitory peptide (cTnl(137-147)) when S23 and S24 are phosphorylated. The inhibitory peptide is also the target of the point mutation cTnI-R145G, which is associated with hypertrophic cardiomyopathy (HCM), a disease associated with sudden death in apparently healthy young adults. It has been shown that both phosphorylation and this mutation alter the cTnC-cTnI (C-I) interaction, which plays a crucial role in modulating contractile activation. However, little is known about the molecular-level events underlying this modulation. Here, we computationally investigated the effects of the cTnI-R145G mutation on the dynamics of cTn, cTnC Ca(2+) handling, and the C-I interaction. Comparisons were made with the cTnI-R145G/S23D/S24D phosphomimic mutation, which has been used both experimentally and computationally to study the cTnI N-terminal specific effects of PKA phosphorylation. Additional comparisons between the phosphomimic mutations and the real phosphorylations were made. For this purpose, we ran triplicate 150 ns molecular dynamics simulations of cTnI-R145G Ca(2+)-bound cTnC(1-161)-cTnI(1-172)-cTnT(236-285), cTnI-R145G/S23D/S24D Ca(2+)-bound cTnC(1-161)-cTnI(1-172)-cTnT(236-285), and cTnI-R145G/PS23/PS24 Ca(2+)-bound cTnC(1-161)-cTnI(1-172)-cTnT(236-285), respectively. We found that the cTnI-R145G mutation did not impact the overall dynamics of cTn, but stabilized crucial Ca(2+)-coordinating interactions. However, the phosphomimic mutations increased overall cTn fluctuations and destabilized Ca(2+) coordination. Interestingly, cTnI-R145G blunted the intrasubunit interactions between the cTnI N-terminal extension and the cTnI inhibitory peptide, which have been suggested to play a crucial role in modulating troponin function during β-adrenergic stimulation. These findings offer a molecular-level explanation for how the HCM mutation cTnI-R145G reduces the modulation of cTn by phosphorylation of S23/S24 during β-adrenergic stimulation.


From surnames to the history of Y chromosomes: the Sardinian population as a paradigm.
A total of 202 Sardinian male subjects were examined for 13 biallelic stable markers, the complex 49a,f/TaqI system and three microsatellites of the Y chromosome in order to investigate, through surname analysis, on a possible territorial heterogeneity inside the island. The study of geographical distribution and linguistic derivation of Sardinian surnames allow us to discover their 'probable place of origin' and reconstruct ancient genetic isolates which borders are, today, no more recognizable. The molecular analysis revealed that about 90% of the Sardinian Y chromosomes fell into haplogroups E-M35, G-M201, I-M26, J-12f2 and R-M269. In contrast with the territorial homogeneity of these haplogroups, when the individuals were distributed according to their birthplace, a significant difference between the three historically and culturally distinct geographical areas into which Sardinia can be subdivided was observed when the individuals were distributed according to the ancestral location of surnames. In particular, the major contribution to this heterogeneity is due to the 'Sardinian-specific' haplogroup I-M26 (almost completely associated with the 49a,f-Ht12/12f2-10Kb/YCAIIa-21/YCAIIb-11 compound haplotype), which shows both a significantly higher incidence in the central-eastern (archaic) area and a significantly lower frequency in the northern area. The results of this study agree with the hypothesis that the ancestral homeland of this specific subset of haplogroup I is the mountainous central-eastern area of Sardinia, where the population underwent a long history of isolation since ancient times, and highlight the informative power of the surname analysis.


Complement factor H-related hybrid protein deregulates complement in dense deposit disease.
The renal disorder C3 glomerulopathy with dense deposit disease (C3G-DDD) pattern results from complement dysfunction and primarily affects children and young adults. There is no effective treatment, and patients often progress to end-stage renal failure. A small fraction of C3G-DDD cases linked to factor H or C3 gene mutations as well as autoantibodies have been reported. Here, we examined an index family with 2 patients with C3G-DDD and identified a chromosomal deletion in the complement factor H-related (CFHR) gene cluster. This deletion resulted in expression of a hybrid CFHR2-CFHR5 plasma protein. The recombinant hybrid protein stabilized the C3 convertase and reduced factor H-mediated convertase decay. One patient was refractory to plasma replacement and exchange therapy, as evidenced by the hybrid protein quickly returning to pretreatment plasma levels. Subsequently, complement inhibitors were tested on serum from the patient for their ability to block activity of CFHR2-CFHR5. Soluble CR1 restored defective C3 convertase regulation; however, neither eculizumab nor tagged compstatin had any effect. Our findings provide insight into the importance of CFHR proteins for C3 convertase regulation and identify a genetic variation in the CFHR gene cluster that promotes C3G-DDD. Monitoring copy number and sequence variations in the CFHR gene cluster in C3G-DDD and kidney patients with C3G-DDD variations will help guide treatment strategies.


Adaptive control of a variable-impedance ankle-foot orthosis to assist drop-foot gait.
An active ankle-foot orthoses (AAFO) is presented where the impedance of the orthotic joint is modulated throughout the walking cycle to treat drop-foot gait. During controlled plantar flexion, a biomimetic torsional spring control is applied where orthotic joint stiffness is actively adjusted to minimize forefoot collisions with the ground. Throughout late stance, joint impedance is minimized so as not to impede powered plantar flexion movements, and during the swing phase, a torsional spring-damper control lifts the foot to provide toe clearance. To assess the clinical effects of variable-impedance control, kinetic and kinematic gait data were collected on two drop-foot participants wearing the AAFO. For each participant, zero, constant, and variable impedance control strategies were evaluated and the results were compared to the mechanics of three age, weight, and height matched normals. We find that actively adjusting joint impedance reduces the occurrence of slap foot allows greater powered plantar flexion and provides for less kinematic difference during swing when compared to normals. These results indicate that a variable-impedance orthosis may have certain clinical benefits for the treatment of drop-foot gait compared to conventional ankle-foot orthoses having zero or constant stiffness joint behaviors.


Risk factors for recurrent emergency department visits for asthma.
Patients presenting with asthma to emergency departments have lost control of their disease, have significant airways obstruction, and frequently require admission to hospital. Although even one visit is not desirable, there is a more disturbing subgroup who repeatedly visit the emergency department. To investigate the reasons for multiple emergency visits, a questionnaire was given to 448 consecutive patients presenting to the two largest adult emergency departments in Ottawa, Canada between November 1989 and April 1991. Within this cohort, those who had made at least three visits in the past year were compared with controls (only one visit in the past year). Although inhaled corticosteroid use increased with multiple visits (indicating increased asthma severity), only 60% of those visiting at least three times in the past year were taking inhaled corticosteroids. Chronic undermedication relative to disease severity was apparent among the cases. The number of visits was associated with nocturnal asthma on a regular basis, work and school absenteeism, frequent visits to their regular physician, and frequent admissions to hospital. Visits were not related to psychological health, environmental allergens/irritants, or lack of perceived asthma severity. The recommendations of current asthma guidelines are not reaching these patients. The issue of translating guidelines from paper to practice must be addressed before highly effective medications can have an important impact on the frequency of emergency department visits.


Selective monoaminergic and histaminergic circuit dysregulation following long-term HIV-1 protein exposure.
Between 30 and 60% of HIV-seropositive individuals develop symptoms of clinical depression and/or apathy. Dopamine and serotonin are associated with motivational alterations; however, histamine is less well studied. In the present study, we used fast-scan cyclic voltammetry in HIV-1 transgenic (Tg) rats to simultaneously analyze the kinetics of nucleus accumbens dopamine (DA), prefrontal cortical serotonin (5-HT), and hypothalamic histamine (HA). For voltammetry, subjects were 15 HIV-1 Tg (7 male, 8 female) and 20 F344/N (11 male, 9 female) adult rats. Both serotonergic and dopaminergic release and reuptake kinetics were decreased in HIV-1 Tg animals relative to controls. In contrast, rates of histamine release and reuptake increased in HIV-1 Tg rats. Additionally, we used immunohistochemical (IHC) methods to identify histaminergic neurons in the tuberomammillary nucleus (TMN) of the hypothalamus. For IHC, subjects were 9 HIV-1 Tg (5 male, 4 female) and 9 F344/N (5 male, 4 female) adult rats. Although the total number of TMN histaminergic cells did not differ between HIV-1 Tg rats and F344/N controls, a significant sex effect was found, with females having an increased number of histaminergic neurons, relative to males. Collectively, these findings illustrate neurochemical alterations that potentially underlie or exacerbate the pathogenesis of clinical depression and/or apathy in HIV-1.


Determination of some in vitro growth requirements of Bacteroides nodosus.
Physical and nutritional factors required for growth of Bacteroides nodosus isolates from ovine foot-rot lesions were examined. Simplified anaerobic culture techniques were devised utilizing a fully soluble, autoclavable, liquid medium (TAS) which contained proteose-peptone, yeast and meat extracts and certain other essential compounds required to promote prompt and serially transferrable growth of cultures from small inocula. The latter included Trypticase, arginine, a reducing agent (most suitably thioglycollic acid) and CO2; serine and Mg2+ markedly increased growth yields. Trypticase could not be replaced by a commercial preparation of acid-hydrolysed casein; other forms of hydrolysed protein gave delayed and inconsistent growth. Maximum growth of cultures required concentrations of 0-02 to 0-35 M-arginine, which could not be replaced by glutamic acid, citrulline or ornithine. Exogenous carbohydrate compounds were not required. The temperature range for optimum growth of cultures was 37 to 39 degrees C, and anaerobic culture conditions were essential for growth and the production of B. nodosus organisms of normal morphology. Solidified TAS media for the isolation and maintenance of B. nodosus cultures were also devised.


Pharmacogenomics of human ABC transporter ABCC11 (MRP8): potential risk of breast cancer and chemotherapy failure.
Some genetic polymorphisms of human ABC transporter genes are reportedly related to the risk of certain diseases and patients' responses to medication. Human ABCC11 functions as an ATP-dependent efflux pump for amphipathic anions. One non-synonymous SNP 538G>A (Gly180Arg) has been found to greatly affect the function and stability of de novo synthesized ABCC11 (Arg180) variant protein. The SNP variant lacking N-linked glycosylation is recognized as a misfolded protein in the endoplasmic reticulum (ER) and readily undergoes proteasomal degradation. This ER-associated degradation of ABCC11 protein underlies the molecular mechanism of affecting the function of apocrine glands. On the other hand, the wild type (Gly180) of ABCC11 is associated with wet-type earwax, axillary osmidrosis, colostrum secretion from the mammary gland, and the potential susceptibility of breast cancer. Furthermore, the wild type of ABCC11 reportedly has ability to efflux cyclic nucleotides and nucleoside-based anticancer drugs. The SNP (538G>A) of the ABCC11 gene is suggested to be a clinical biomarker for prediction of chemotherapeutic efficacy. Major obstacle to the successful chemotherapy of human cancer is development of resistance, and nucleoside-based chemotherapy is often characterized by inter-individual variability. This review provides an overview about the discovery and the genetic polymorphisms in human ABCC11. Furthermore we focus on the impact of ABCC11 538G>A on the apocrine phenotype, patients' response to nucleoside-based chemotherapy, and the potential risk of breast cancer.


A Flow-Based Model of the HIV Care Continuum in the United States.
Understanding the flow of patients through the continuum of HIV care is critical to determine how best to intervene so that the proportion of HIV-infected persons who are on antiretroviral treatment and virally suppressed is as large as possible. Using immunological and virological data from the Centers for Disease Control and Prevention and the North American AIDS Cohort Collaboration on Research and Design from 2009 to 2012, we estimated the distribution of time spent in and dropout probability from each stage in the continuum of HIV care. We used these estimates to develop a queueing model for the expected number of patients found in each stage of the cascade. HIV-infected individuals spend an average of about 3.1 months after HIV diagnosis before being linked to care, or dropping out of that stage of the continuum with a probability of 8%. Those who link to care wait an additional 3.7 months on average before getting their second set of laboratory results (indicating engagement in care) or dropping out of care with probability of almost 6%. Those engaged in care spent an average of almost 1 year before achieving viral suppression on antiretroviral therapy or dropping out with average probability 13%. For patients who achieved viral suppression, the average time suppressed on antiretroviral therapy was an average of 4.5 years. Interventions should be targeted to more rapidly identifying newly infected individuals, and increasing the fraction of those engaged in care that achieves viral suppression.


Eliminating hotspots in a multi-chip LED array direct backlight system with optimal patterned reflectors for uniform illuminanceand minimal system thickness.
We propose an optical design process that significantly reduces the time and costs in direct backlight unit (BLU) development. In it, the basic system specifications are derived from the optical characteristics of RGB light-emitting diodes (LEDs) comprising the BLU. The driving currents are estimated to determine the theoretical RGB flux ratio for a desired white point. The number of LEDs needed to produce the target luminance is then calculated from the combined optical efficiencies of the components. Last, an appropriate array configuration is sought based on the illuminance distribution function for meeting the target uniformity. To showcase the design process we built two 42-inch triangular cluster arrays of 40 x 16 LED elements. When a flat reflective sheet was used, the minimum thickness required of the system to satisfy the target uniformity was 30 mm. Introducing a patterned reflective sheet removed hotspots that resulted from reducing the system thickness without the aid of additional optical components. Using an optimized patterned reflective sheet, reduction in system thickness as much as 5 mm was possible.


Phase diagram of restricted Boltzmann machines and generalized Hopfield networks with arbitrary priors.
Restricted Boltzmann machines are described by the Gibbs measure of a bipartite spin glass, which in turn can be seen as a generalized Hopfield network. This equivalence allows us to characterize the state of these systems in terms of their retrieval capabilities, both at low and high load, of pure states. We study the paramagnetic-spin glass and the spin glass-retrieval phase transitions, as the pattern (i.e., weight) distribution and spin (i.e., unit) priors vary smoothly from Gaussian real variables to Boolean discrete variables. Our analysis shows that the presence of a retrieval phase is robust and not peculiar to the standard Hopfield model with Boolean patterns. The retrieval region becomes larger when the pattern entries and retrieval units get more peaked and, conversely, when the hidden units acquire a broader prior and therefore have a stronger response to high fields. Moreover, at low load retrieval always exists below some critical temperature, for every pattern distribution ranging from the Boolean to the Gaussian case.


Cost-effectiveness of different strategies to monitor adults on antiretroviral treatment: a combined analysis of three mathematical models.
WHO's 2013 revisions to its Consolidated Guidelines on antiretroviral drugs recommend routine viral load monitoring, rather than clinical or immunological monitoring, as the preferred monitoring approach on the basis of clinical evidence. However, HIV programmes in resource-limited settings require guidance on the most cost-effective use of resources in view of other competing priorities such as expansion of antiretroviral therapy coverage. We assessed the cost-effectiveness of alternative patient monitoring strategies. We evaluated a range of monitoring strategies, including clinical, CD4 cell count, and viral load monitoring, alone and together, at different frequencies and with different criteria for switching to second-line therapies. We used three independently constructed and validated models simultaneously. We estimated costs on the basis of resource use projected in the models and associated unit costs; we quantified impact as disability-adjusted life years (DALYs) averted. We compared alternatives using incremental cost-effectiveness analysis. All models show that clinical monitoring delivers significant benefit compared with a hypothetical baseline scenario with no monitoring or switching. Regular CD4 cell count monitoring confers a benefit over clinical monitoring alone, at an incremental cost that makes it affordable in more settings than viral load monitoring, which is currently more expensive. Viral load monitoring without CD4 cell count every 6-12 months provides the greatest reductions in morbidity and mortality, but incurs a high cost per DALY averted, resulting in lost opportunities to generate health gains if implemented instead of increasing antiretroviral therapy coverage or expanding antiretroviral therapy eligibility. The priority for HIV programmes should be to expand antiretroviral therapy coverage, firstly at CD4 cell count lower than 350 cells per μL, and then at a CD4 cell count lower than 500 cells per μL, using lower-cost clinical or CD4 monitoring. At current costs, viral load monitoring should be considered only after high antiretroviral therapy coverage has been achieved. Point-of-care technologies and other factors reducing costs might make viral load monitoring more affordable in future. Bill & Melinda Gates Foundation, WHO.


Providing Diverse Trainees an Early and Transparent Introduction to Academic Appointment and Promotion Processes.
The growth in number of medical schools and increased numbers of faculty tracks have combined with evolving criteria for promotion to trigger a call for greater transparency of academic appointment and promotion processes. Most vulnerable to confusion about these changes are first-generation and diverse medical students and residents, the upstream pipeline of the academic medicine workforce. Diverse medical students have expressed diminished interest in academia because of perceived obstacles in appointment and promotion processes. This workshop was designed to utilize didactics and career reflection exercises to help trainees learn: (1) how to define core terms related to academic appointment and promotion processes, (2) how to compare data elements for different CVs and portfolios, (3) common steps in submitting a promotion package, and (4) that they can immediately begin to document content for academic CVs, portfolios, and promotion packages. One hundred forty-five diverse participants completed an evaluation at eight conferences across the U.S. More than 90% strongly agreed or agreed that the aforementioned objectives were met. Participants commented that the workshop was "illuminating," was "very informative," and "provided an inside look of how faculty are evaluated." Results showed an immediate impact on participants' self-reported confidence to negotiate appointment and promotion processes. Increases in self-rated confidence to negotiate appointment and promotion processes may help sustain trainees' interest in becoming future faculty. Further monitoring will be needed to determine if early exposure to these concepts improves probability of seeking, obtaining, and maintaining appointments.


Comparing the Healthcare Utilization and Engagement in a Sample of Transgender and Cisgender Bisexual+ Persons.
People who identify as non-monosexual and transgender experience disparities in engagement with healthcare services relative to monosexual and cisgender persons, respectively. However, little is known about the healthcare utilization of those with intersecting sexual and gender minority identities. We explored the knowledge, attitudes, and health motivation of non-monosexually identified transgender participants regarding preventive care and access to sexual healthcare services. We surveyed 87 ciswomen, 34 transwomen, and 27 transmen, all of whom identified as bisexual, pansexual, or queer (bi+). We assessed their access to health care, health outcome experiences, confidence with talking about anogenital topics, proactivity toward their health, comfort with healthcare providers, and knowledge about HPV and examined differences across groups. The data indicated that bi+ transmen and transwomen were more likely to be uninsured or on a government-sponsored insurance plan relative to bi+ ciswomen. Only a minority of transmen and transwomen had seen an obstetrician/gynecologist compared with ciswomen. Transmen were less likely to have received a pelvic examination or cervical Pap smear in their lifetime. Transgender participants had significantly less correct knowledge about HPV relative to ciswomen. Finally, relative to ciswomen, transgender participants reported lower comfort talking with health providers. Our findings suggest that bi+ transmen and transwomen access care less than bi+ ciswomen and have less health knowledge and comfort with their providers. Implications for intervention include encouraging transgender individuals to seek routine screenings, reducing structural barriers to care based on medical coverage, and improving patient-provider competencies around bi+ and transgender health needs.


Foodborne parasites: Outbreaks and outbreak investigations. A meeting report from the European network for foodborne parasites (Euro-FBP).
Foodborne parasites (FBP) are a neglected topic in food safety, due to lack of awareness of their importance for public health, their biological diversity, and, for many FBP, lack of standardized detection methods, which complicates identification of the infection vehicle. The COST Action FA1408, A European Network for Foodborne Parasites (Euro-FBP), aims to limit the impact of FBP on public health by facilitating multidisciplinary cooperation and partnership among researchers, and between researchers and stakeholders. Outbreaks represent a common topic overarching specialization in one or more FBP, thus providing a platform for knowledge exchange. This report summarizes the outcomes of a meeting within the Euro-FBP consortium entitled 'Outbreaks and Outbreak Investigations'. Recent and historical outbreaks of trichinellosis, opisthorchiasis, and cryptosporidiosis were used as examples to underline the complexity of the topic, the different foods implicated and their traceability, and the lack of standardized detection methods for some parasites. Possible solutions to overcome current limitations were also illustrated. The meeting provided an opportunity to learn from recent advances in the study of bacterial foodborne outbreaks, with an emphasis on genome analysis.


Evaluating the clinical and physiological effects of long term ultraviolet B radiation on guinea pigs (Cavia porcellus).
Vitamin D is an important hormone in vertebrates. Most animals acquire this hormone through their diet, secondary to exposure to ultraviolet B (UVB) radiation, or a combination thereof. The objectives for this research were to evaluate the clinical and physiologic effects of artificial UVB light supplementation on guinea pigs (Cavia porcellus) and to evaluate the long-term safety of artificial UVB light supplementation over the course of six months. Twelve juvenile acromelanic Hartley guinea pigs were randomly assigned to one of two treatment groups: Group A was exposed to 12 hours of artificial UVB radiation daily and Group B received only ambient fluorescent light for 12 hours daily. Animals in both groups were offered the same diet and housed under the same conditions. Blood samples were collected every three weeks to measure blood chemistry values, parathyroid hormone, ionized calcium, and serum 25-hydroxyvitamin D3 (25-OHD3) levels. Serial ophthalmologic examinations, computed tomography scans, and dual energy x-ray absorptiometry scans were performed during the course of the study. At the end of the study the animals were euthanized and necropsied. Mean ± SD serum 25-OHD3 concentrations differed significantly in the guinea pigs (p<0.0001) between the UVB supplementation group (101.49±21.81 nmol/L) and the control group (36.33±24.42 nmol/L). An increased corneal thickness in both eyes was also found in the UVB supplementation compared to the control group (right eye [OD]: p<0.0001; left eye [OS]: p<0.0001). There were no apparent negative clinical or pathologic side effects noted between the groups. This study found that exposing guinea pigs to UVB radiation long term significantly increased their circulating serum 25-OHD3 levels, and that this increase was sustainable over time. Providing guinea pigs exposure to UVB may be an important husbandry consideration that is not currently recommended.


Chemical imaging of microstructures of plant tissues within cellular dimension using synchrotron infrared microspectroscopy.
Synchrotron radiation-based Fourier transform infrared microspectroscopy (SR-FTIR) is an advanced bioanalytical technique capable of exploring the chemistry within microstructures of plant and animal tissues with a high signal to noise ratio at high ultraspatial resolutions (3-10 microm) without destruction of the intrinsic structures of a tissue. This technique is able to provide information relating to the quantity, composition, structure, and distribution of chemical constituents and functional groups in a tissue. The objective of this study was to illustrate how the SR-FTIR technique can be used to image inherent structures of plant tissues on a cellular level (pixel size, approximately 10 microm x 10 microm). The results showed that with the extremely bright synchrotron light, spectra with high signal to noise ratios were obtained from areas as small as 10 microm x 10 microm in the plant tissue, which allowed us to "see" plant tissue in a chemical sense on a cellular level. The ultraspatial resolved imaging of plant tissues by stepping in pixel-sized increments was obtained. Chemical distributions of plant tissues such as lignin, cellulose, protein, lipid, and total carbohydrate could be mapped. These images revealed the chemical information of plant intrinsic structure. In conclusion, SR-FTIR can provide chemical and functional characteristics of plant tissue at high ultraspatial resolutions. The SR-FTIR microspectroscopic images can generate spatially localized functional group and chemical information within cellular dimensions.


Exploring the influence of steric, electronic and lipophilic descriptors of 1,3-diarly propenones on their anti-inflammatory activity.
Various compounds from natural and synthetic origins containing the 1,3-diarylpropenone structure have been reported to produce a variety of biological activities like anti-microbial, anti-inflammatory, vascular muscle relaxant, etc. A systematic analysis of the structural features responsible for anti-inflammatory activity and a possible mode of their actions were proposed to be evaluated by synthesizing a set of compounds, screening them for anti-inflammatory activity and developing a QSAR model. Two types of 1,3-diarylpropenone derivatives were synthesized employing the Claisen-Schmidt condensation. These compounds were then screened for their in vivo anti- inflammatory activity by the carrageenin induced rat paw edema method and also for in vitro cyclooxygenase-2 (COX-2) inhibition activity using a colorimetric kit for COX (ovine) inhibitor screening assay. These derivatives and their anti-inflammatory activity data were employed for QSAR analysis on Vlife MDS 3.5 software. The molecules were divided into training and test sets based on observed activity and QSAR models were generated for the training set and validated. The activity of the molecules of the test set was predicted according to the QSAR equation fit. Possible correlation between observed anti-inflammatory activity and in vitro cyclooxygenase-2 inhibition was also studied. Insignificant difference between the observed and predicted biological activity revealed that the selected electronic, steric and lipophilic parameters have a significant correlation (r(2)=0.85) with anti-inflammatory activity of the selected class of compounds. On the basis of results it may be suggested that the 1,3-diaryl-2-propen-1-ones framework is an attractive template for structural optimization to achieve better potency of anti-inflammatory activity. Similarly, the relatively low correlation between anti-inflammatory activity and cyclooxygenase-2 inhibition indicates that other modes of actions may also be responsible for the anti-inflammatory activity of the tested compounds.


Regulation of eosinophil-active cytokine production from human cord blood-derived mast cells.
Human mast cells are multifunctional tissue-dwelling cells that play a crucial role in eosinophil-dependent disorders, such as asthma and parasitic diseases, by the secretion of eosinophil-active mediators. Mast cell-derived cytokines, generated in response to cross-linking of the high-affinity IgE receptor, can regulate eosinophil activation, survival, and chemotaxis. In this study, mast cells generated from human cord blood progenitors (stem cells) were studied for eosinophil-active inflammatory cytokine expression. Cord blood-derived mast cells (CBDMC) expressed typical intracellular scroll granules and microvilli-like structures on their cell surfaces, demonstrated the presence of tryptase, and elaborated prostaglandin D2 (PGD2) after cross-linkage of the high-affinity receptor for IgE (FcepsilonRI). CBDMC expressed tumor necrosis factor-alpha (TNF-alpha) and the eosinophil-active growth factors, interleukin-5 (IL-5) and granulocyte-macrophage colony-stimulating factor (GM-CSF) after activation. (IL-1beta greatly enhanced IgE-dependent production of these cytokines in response to FcepsilonRI cross-linkage, suggesting a role for bystander/phagocytic cells in modulating mast cell function. In contrast, interferon-alpha (IFN-alpha) inhibited IL-5 and GM-CSF generation, and the glucocorticoid, dexamethasone (Dex), inhibited production of IL-5 and GM-CSF from CBDMC. A macrophage-mast cell-eosinophil axis may exist in vivo that may be susceptible to pharmacologic manipulation.


Receptor activator of nuclear factor-κB ligand and sclerostin expression in osteocytes of alveolar bone in rats with ligature-induced periodontitis.
Osteocytes are increasingly recognized as significant sources of osteoclast differentiation factor, receptor activator of nuclear factor-κB ligand (RANKL), and osteoblast differentiation inhibitory factor, sclerostin. In this study, RANKL and sclerostin expression of osteocytes is investigated in rats with ligature-induced periodontitis. Rats were divided into control and periodontitis groups, and periodontitis was induced by ligature on the mandibular first molars. At 1, 3, 10, and 20 days after ligature, histologic analyses of alveolar bone (AB) and osteoid areas in the molar furcation were performed. The numbers of osteoclasts and RANKL- and sclerostin-positive osteocytes were estimated by tartrate-resistant acid phosphatase staining and immunohistochemistry, respectively. The AB area gradually decreased at day 10 after ligature and increased at day 20. The number of osteoclasts markedly increased at day 3 and then decreased. Conversely, osteoid formation was suppressed up to day 3 and then showed a remarkable increase above control level at day 20. The number of RANKL-positive osteocytes increased at days 1 and 3 and then decreased. Sclerostin-positive osteocytes markedly increased at days 3 and 10 but decreased below control level at day 20. These results show that AB loss is accompanied by enhanced osteoclast formation and suppressed osteoid formation. Osteocytes express RANKL when osteoclast formation increases, and they express sclerostin when osteoid formation is suppressed. Conversely, osteocytic sclerostin expression decreases when osteoid formation increases. These findings suggest that osteocytes may be important in AB loss via RANKL and sclerostin expression in periodontitis.


First Report of Peanut Pod Rot Caused by Neocosmospora vasinfecta in Northern China.
From 2006 to 2010, peanut (Arachis hypogaea) pod rot became more prevalent in northern China, especially in the Sha River drainage area. The incidence of pod rot ranged from 30 to 100%. Typical symptoms were black rot of the pods, but no obvious morphological abnormality of the aboveground parts of infected plants was observed. Brown or black spots appeared on many pods when initially infected and then all peanut pods became black and rotten. The same fungus was isolated from 54 surface-disinfested lesions (85.2% of all lesions) on potato dextrose agar (PDA) media. One isolate, designated as HBXLb, was chosen for further characterization. In culture, both anamorph and teleomorph were present. Mycelia of the fungus grew quickly (colonies were 3.2 cm in diameter in 3 days) and became white and floccose on PDA at 28°C. The hyaline, elongated-to-cylindrical conidia aggregated on the slimy heads of conidiogenous cells that developed on undifferentiated hyphae after incubation for 3 to 4 days. Conidial sizes varied from 5 to 10 × 1.5 to 3 μm (n = 50) and were mostly single celled. Some conidia appeared slightly curved. The morphology was consistent with Acremonium spp. Numerous ascomata (perithecia) formed within 10 to 14 days when incubated at 28°C under light and dark conditions. Perithecia were orange-brown, strawberry shaped (300 to 400 μm in diameter), and ostiolate on the top. Cylindrical asci, with an average size of 90 to 110 × 7.5 to 9 μm, were present inside the ascomata with each containing eight ascospores in a row. The ascospores were brownish, spherical to ellipsoidal, and 10 to 15 × 8 to 12 μm. The cultural and morphological characteristics of isolate HBXLb matched the description of N. vasinfecta (2). The internal transcribed spacer (ITS) region of rDNA was amplified by the primer pairs ITS4/ITS5. A 525-bp amplicon (ITS4-5.8s-ITS5) was obtained and sequenced (GenBank Accession No. HM461901). The ITS sequence was a 100% match to N. vasinfecta strain N-JXLN01 (GenBank Accession No. GU213063) by BLASTn in GenBank. Pathogenicity tests were conducted on detached pods of peanut cultivar Jihua 4. Forty surface-disinfested peanut pods were soaked in a conidial suspension (105 conidia per ml) for 2 min and 40 pods were soaked in sterile water as a control. Then all peanut pods were maintained in moist petri dishes under darkness for 14 days at 28°C. Brown or black spots appeared on all pods inoculated with the fungus within 10 days, while the controls remained healthy. Symptoms were similar to those originally observed in the field, and N. vasinfecta could be reisolated from all infected pods. This fungus previously has been reported as the pathogen of foot rot of peanut in South Africa (1), Taiwan (4), and Australia (3). To our knowledge, this is the first report of peanut pod rot caused by N. vasinfecta in China. References: (1) S. W. Baard et al. Phytophylactica 17:49, 1985. (2) O. A. Cornely et al. Emerg. Infect. Dis. 7:149, 2001. (3) M. F. Fuhlbohm et al. Australas. Plant Dis. Notes 2:3, 2007. (4) J. W. Huang et al. Plant Pathol. Bull. 1:203, 1992.


Unbiased Cell-based Screening in a Neuronal Cell Model of Batten Disease Highlights an Interaction between Ca2+ Homeostasis, Autophagy, and CLN3 Protein Function.
Abnormal accumulation of undigested macromolecules, often disease-specific, is a major feature of lysosomal and neurodegenerative disease and is frequently attributed to defective autophagy. The mechanistic underpinnings of the autophagy defects are the subject of intense research, which is aided by genetic disease models. To gain an improved understanding of the pathways regulating defective autophagy specifically in juvenile neuronal ceroid lipofuscinosis (JNCL or Batten disease), a neurodegenerative disease of childhood, we developed and piloted a GFP-microtubule-associated protein 1 light chain 3 (GFP-LC3) screening assay to identify, in an unbiased fashion, genotype-sensitive small molecule autophagy modifiers, employing a JNCL neuronal cell model bearing the most common disease mutation in CLN3. Thapsigargin, a sarco/endoplasmic reticulum Ca(2+)-ATPase (SERCA) Ca(2+) pump inhibitor, reproducibly displayed significantly more activity in the mouse JNCL cells, an effect that was also observed in human-induced pluripotent stem cell-derived JNCL neural progenitor cells. The mechanism of thapsigargin sensitivity was Ca(2+)-mediated, and autophagosome accumulation in JNCL cells could be reversed by Ca(2+) chelation. Interrogation of intracellular Ca(2+) handling highlighted alterations in endoplasmic reticulum, mitochondrial, and lysosomal Ca(2+) pools and in store-operated Ca(2+) uptake in JNCL cells. These results further support an important role for the CLN3 protein in intracellular Ca(2+) handling and in autophagic pathway flux and establish a powerful new platform for therapeutic screening.


Low affinity binding of plasma proteins to lipid-coated quantum dots as observed by in situ fluorescence correlation spectroscopy.
Protein binding to lipid-coated nanoparticles has been pursued quantitatively by using fluorescence correlation spectroscopy. The binding of three important plasma proteins to lipid-enwrapped quantum dots (QDs) shows very low affinity, with an apparent dissociation coefficient in the range of several hundred micromolar. Thus, the tendency to adsorb is orders of magnitude weaker than for QDs coated with dihydrolipoic acid.


Rainwater harvesting: quality assessment and utilization in The Netherlands.
The use of roof-collected rainwater as a freely available and sustainable alternative to drinking water produced by drinking water companies increases worldwide. Initially, rainwater is free of microbial contamination, but it may become contaminated by animals and humans or, alternatively, human pathogens may grow in stored rainwater resulting in a significant human health risk from infectious diseases. This three-year study demonstrated that rainwater stored in different reservoirs in The Netherlands was frequently faecally contaminated and incidentally contained potential human pathogens such as Campylobacter, Cryptosporidium, Giardia, Aeromonas hydrophila and Legionella. Analysis of samples during a period with variable weather conditions showed a correlation between rainfall intensity and faecal indicator counts and increased detection of pathogens after heavy rainfall incidents. Outside temperature had a limited effect on both the temperature and the microbiological quality of the water in the reservoirs, which did not comply with Dutch drinking water legislation and should thus not be consumed without treatment. In general, a health risk may arise from exposure to pathogens when contaminated droplets are inhaled, ingested or come into contact with the skin. Health risks may be reduced by regular cleaning of the collection, storage and transport means, but to assess their efficacy field intervention studies are required.


Single-step purification of intrinsic protein complexes in Saccharomyces cerevisiae using regenerable calmodulin resin.
Characterization of protein- protein interactions is a vital aspect of molecular biology as multi-protein complexes are the functional units of cellular processes and defining their interactions provides valuable insights into their function based on a "guilt by association" concept. To identify the components in complexes, purification of the latter near homogeneity is required. The tandem affinity purification (i.e., TAP) method, coupled with mass spectrometry have been extensively used to define native protein complexes and transient protein-protein interactions under near physiological conditions (i.e., conditions approximate of the internal milieu) in Saccharomyces cerevisiae. Generally, TAP consists of two-stage protein enrichment using dual affinity tags, a calmodulin-binding peptide and a Staphylococcus aureus protein-A, separated by a tobacco etch virus protease site, which are fused to either the C- or N-terminal of the target protein. TAP-tagging has proved to be a powerful method for studying functional relationship between proteins and generating large-scale protein networks. The method described in this paper provides an inexpensive single-step purification alternative to the traditional two step affinity purification of TAP-tagged proteins using only the calmodulin-binding peptide affinity tag. Moreover, a novel protocol for the regeneration of the calmodulin-agarose resin is outlined and validated. This basic approach allows fast and cost-effective purification of proteins and their interacting partners from Saccharomyces cerevisiae.


(-)-Gossypol enhances response to radiation therapy and results in tumor regression of human prostate cancer.
Radioresistance markedly impairs the efficacy of tumor radiotherapy and involves antiapoptotic signal transduction pathways that prevent radiation-induced cell death. The majority of human prostate cancers overexpress the important antiapoptotic proteins Bcl-2 and/or Bcl-xL, which render tumors resistant to radiation therapy. (-)-Gossypol, a natural polyphenol product from cottonseed, has recently been identified as a potent small molecule inhibitor of both Bcl-2 and Bcl-xL. In the current study, we investigated the antitumor activity of (-)-gossypol in prostate cancer and tested our hypothesis that (-)-gossypol may improve prostate cancer's response to radiation by potentiating radiation-induced apoptosis and thus making cancer cells more sensitive to ionizing radiation. Our data show that (-)-gossypol potently enhanced radiation-induced apoptosis and growth inhibition of human prostate cancer PC-3 cells, which have a high level of Bcl-2/Bcl-xL proteins. Our in vivo studies using PC-3 xenograft models in nude mice show that orally given (-)-gossypol significantly enhanced the antitumor activity of X-ray irradiation, leading to tumor regression in the combination therapy. In situ terminal deoxynucleotidyl transferase-mediated nick end labeling staining showed that significantly more apoptotic cells were induced in the tumors treated with (-)-gossypol plus radiation than either treatment alone. Anti-CD31 immunohistochemical staining indicates that (-)-gossypol plus radiation significantly inhibited tumor angiogenesis. Our results show that the natural polyphenol inhibitor of Bcl-2/Bcl-xL, (-)-gossypol, can radiosensitize prostate cancer in vitro and in vivo without augmenting toxicity. (-)-Gossypol may improve the outcome of current prostate cancer radiotherapy and represents a promising novel anticancer regime for molecular targeted therapy of hormone-refractory prostate cancer with Bcl-2/Bcl-xL overexpression.


Informal caregiving for dementia patients: the contribution of patient characteristics and behaviours to caregiver burden.
The burden often associated with informal caregiving for patients with dementia is associated with negative effects on health, both physiologically and in terms of caregiver cognition. There is wide variation in the level of burden experienced by dementia caregivers. To better understand caregiver burden, it is thus important to understand the factors associated with level of burden. In the current study, we collected carer burden and putative associated factors at baseline, 6 and 12 months. Hierarchical regression was used to assess the impact of these factors on caregiver burden. We assessed self-reported carer burden, patient behavioural and safety issues, and level of difficulty associated with providing assistance with activities of daily living (ADL). Patients' age was also recorded, and trained nurses assessed patient cognitive performance using the quick mild cognitive impairment screen. At baseline, patients' age, cognition and ADLs were associated with burden, and safety and challenging behaviour were both significantly associated with burden independent of the other factors. Change in burden was associated with change in carer-reported safety at 6-month follow-up, and with change in safety and change in carer-reported challenging behaviours at 12-month follow-up. Safety issues and challenging behaviours are associated with carer burden, even after accounting for cognitive and functional impairment in the person with dementia. As dementia progresses, monitoring these factors may help to inform stress-management strategies for caregivers.


Preterm infant massage therapy research: a review.
In this paper, preterm infant massage therapy studies are reviewed. Massage therapy has led to weight gain in preterm infants when moderate pressure massage was provided. In studies on passive movement of the limbs, preterm infants also gained significantly more weight, and their bone density also increased. Research on ways of delivering the massage is also explored including using mothers versus therapists and the added effects of using oils. The use of mothers as therapists was effective in at least one study. The use of oils including coconut oil and safflower oil enhanced the average weight gain, and the transcutaneous absorption of oil also increased triglycerides. In addition, the use of synthetic oil increased vagal activity, which may indirectly contribute to weight gain. The weight gain was associated with shorter hospital stays and, thereby, significant hospital cost savings. Despite these benefits, preterm infant massage is only practiced in 38% of neonatal intensive care units. This may relate to the underlying mechanisms not being well understood. The increases noted in vagal activity, gastric motility, insulin and IGF-1 levels following moderate pressure massage are potential underlying mechanisms. However, those variables combined do not explain all of the variance in weight gain, highlighting the need for additional mechanism studies.


First-degree relatives of persons with type 1 diabetes: insulin resistance and enterovirus infection are associated with different patterns of islet cell autoimmunity.
Type 1 diabetes (T1D) results from the interaction of genetic and environmental factors. Previous studies indicate an association between detection of Enterovirus (EV) genome in blood and the clinical onset of T1D. Insulin resistance can also represent a risk factor for progression to clinically overt T1D. This study aimed at evaluating whether there is association between both EV infection and insulin resistance with islet autoantibodies in first-degree relatives of persons with type 1 diabetes. We collected sera from 94 first-degree relatives with (32) or without (64) islet cell antibodies (ICA) from the Cuban T1D prediction program. Blood glucose and insulin concentrations were determined. Antibodies to GAD65 and IA-2 were determined by radioimmunoassay. Insulin resistance was estimated by the homeostasis model assessment (HOMA-IR). EV-RNA was detected in serum using a highly sensitive reverse transcriptase-polymerase chain reaction method. The occurrence of EV-RNA was higher in ICA-positive relatives than in ICA-negative ones [15.6% (5/32) vs. 1.6% (1/62), P = 0.016]. GAD65 autoantibodies were more frequent in subjects with insulin resistance [34.5% (10/29) vs. 13.9% (9/65), P = 0.028] as defined by the HOMA-IR value. GAD65 autoantibodies also positively correlated with HOMA-IR (r.bis = 0.28, P < 0.01). IA-2 autoantibodies did correlate neither with EV-RNA nor with insulin resistance. There was no association between the presence of EV-RNA and insulin resistance. Our data suggest that enterovirus infection and insulin resistance are two independent events associated with ICA and GAD65 autoantibodies, respectively. These observations support the multifactorial nature of T1D.


Defining the complications of cryoablation and radio frequency ablation of small renal tumors: a multi-institutional review.
Ablative treatments (cryoablation or radio frequency ablation) for renal cell carcinoma aim to decrease morbidity by treating renal tumors in situ, eliminating the need for extirpation. These technologies have potential for complications previously unassociated with renal tumor treatment. We identified complications associated with percutaneous and laparoscopic ablative treatment of renal tumors. Groups at medical centers with reported experience with ablation of renal tumors were invited to participate in this study. Each group submitted retrospective data regarding overall ablative treatment experience and associated complications. For each incident the nature of the complication, its associated morbidity, the necessity and nature of any subsequent interventions, and the final patient outcome were evaluated. Complications were divided into minor and major categories. Data were collected from groups at 4 institutions with a combined experience of 271 cases. Of these cases 139 were cryoablation and 133 were radio frequency ablation. There were 181 procedures performed percutaneously and 90 performed laparoscopically. A total of 30 complications occurred (11.1%) with 5 major (1.8%) and 25 minor (9.2%) complications, and 1 death (0.4%). Overall 26 of the 30 complications (86.7%) were directly attributable to the ablation procedure. The most common complication was pain or paraesthesia at the probe insertion site. Ablation technologies appear to have a low complication profile when used to treat small renal tumors. The majority of complications are minor and require observation only. Further study and followup are necessary to determine long-term oncological efficacy.


Assuring the research competence of orthopedic graduates.
To assure that orthopaedic residents have the knowledge needed to critically appraise the medical/surgical literature and the skills needed to design and conduct musculoskeletal and orthopaedic research. Residency program of the Department of Orthopaedic Surgery, Sports Medicine and Rehabilitation at Wright State University Boonshoft School of Medicine, Dayton, Ohio. Established in 2005, members of the residency program's Research Team (RT) include orthopaedic faculty and residents, university faculty and graduate students, community scientists, and department staff. The membership and responsibilities of the Research Team are described. The goals of the three-month research rotation during the second year of residency and the activities of the assigned resident are reported. The work of the two research residents during the second year of their six-year program is explained. Helpful educational resources are noted, the role of the faculty research mentor defined, funding sources listed, and the financial support for RT members and research staff presented. The scholarly activity of orthopaedic surgery residents increased substantially from the five-year period before the implementation of the Research Team (September 2000 through August 2005) to the four-year period after initiation of the RT (September 2005 through November 2009). For peer-reviewed scholarship activity, publications on which residents were authors increased from 1 to 10 per year, national presentations at professional meetings increased from 6 to 11 per year, and local and regional presentations increased from 2 to 4 per year. The Research Team is an effective strategy for assuring that orthopaedic residents have the knowledge and skills to evaluate clinical care evidence and to conduct musculoskeletal and orthopaedic research.


Downregulation of adipose glutathione S-transferase A4 leads to increased protein carbonylation, oxidative stress, and mitochondrial dysfunction.
Peripheral insulin resistance is linked to an increase in reactive oxygen species (ROS), leading in part to the production of reactive lipid aldehydes that modify the side chains of protein amino acids in a reaction termed protein carbonylation. The primary enzymatic method for lipid aldehyde detoxification is via glutathione S-transferase A4 (GSTA4) dependent glutathionylation. The objective of this study was to evaluate the expression of GSTA4 and the role(s) of protein carbonylation in adipocyte function. GSTA4-silenced 3T3-L1 adipocytes and GSTA4-null mice were evaluated for metabolic processes, mitochondrial function, and reactive oxygen species production. GSTA4 expression in human obesity was evaluated using microarray analysis. GSTA4 expression is selectively downregulated in adipose tissue of obese insulin-resistant C57BL/6J mice and in human obesity-linked insulin resistance. Tumor necrosis factor-alpha treatment of 3T3-L1 adipocytes decreased GSTA4 expression, and silencing GSTA4 mRNA in cultured adipocytes resulted in increased protein carbonylation, increased mitochondrial ROS, dysfunctional state 3 respiration, and altered glucose transport and lipolysis. Mitochondrial function in adipocytes of lean or obese GSTA4-null mice was significantly compromised compared with wild-type controls and was accompanied by an increase in superoxide anion. These results indicate that downregulation of GSTA4 in adipose tissue leads to increased protein carbonylation, ROS production, and mitochondrial dysfunction and may contribute to the development of insulin resistance and type 2 diabetes.


Microbial production of vitamin B12.
One of the most alluring and fascinating molecules in the world of science and medicine is vitamin B12 (cobalamin), which was originally discovered as the anti pernicious anemia factor and whose enigmatic complex structure is matched only by the beguiling chemistry that it mediates. The biosynthesis of this essential nutrient is intricate, involved and, remarkably, confined to certain members of the prokaryotic world, seemingly never have to have made the eukaryotic transition. In humans, the vitamin is required in trace amounts (approximately 1 microg/day) to assist the actions of only two enzymes, methionine synthase and (R)-methylmalonyl-CoA mutase; yet commercially more than 10 t of B12 are produced each year from a number of bacterial species. The rich scientific history of vitamin B12 research, its biological functions and the pathways employed by bacteria for its de novo synthesis are described. Current strategies for the improvement of vitamin B12 production using modern biotechnological techniques are outlined.


Spatial control of neuronal cell attachment and differentiation on covalently patterned laminin oligopeptide substrates.
The spatial control of neuronal cell attachment and differentiation via specific receptor mediated interactions, may provide an effective means for the in vitro reconstruction of neuronal cell architecture. In this study, receptor-specific oligopeptide sequences derived from the extracellular matrix (ECM) molecule laminin, a potent neural cell attachment and differentiation promoter were covalently bound on fluorinated ethylene propylene (FEP) films. The degree of receptor-specific cell attachment and the ability to spatially control neurite outgrowth by covalently patterning the oligopeptide sequences on the FEP film surface were assessed. FEP films were first chemically activated with a Radio Frequency Glow Discharge (RFGD) process that covalently replaces the surface fluorine atoms with reactive hydroxyl groups. Oligopeptides containing the YIGSR sequence from the B1 chain of laminin and the water soluble oligopeptide containing the IKVAV sequence (CSRARKQAASIKVAVSADR) from the A chain were covalently bound to the hydroxylated FEP films. Electron Spectroscopy for Chemical Analysis (ESCA) verified the covalent attachment of the oligopeptides to the material surface. The degree of receptor mediated NG108-15 cell attachment on immobilized CDPGYIGSR films was determined using competitive binding media. A 78% reduction in cell attachment was observed on films containing CDPGYIGSR in the cell plating medium. Only a 23% reduction in cell attachment was noted on films plated in medium containing a mock CDPGYIGSK sequence. FEP films immobilized with the IKVAV oligopeptide sequence were shown to mediate PC12 cell attachment and a competitive binding medium also significantly attenuated cell attachment on the immobilized films. The spatial patterning of these oligopeptide sequences to the FEP surface was shown to localize cell attachment and neurite extension on the patterned pathways. The surrounding unmodified FEP surface was inhibitory in serum containing medium and prevented cellular interactions outside the oligopeptide modifications. The spatial immobilization of laminin oligopeptides on FEP films provides a means to organize the attachment and differentiation of neuronal cells in a receptor-specific manner.


Antibiotic resistance determinants in Acinetobacter spp and clinical outcomes in patients from a major military treatment facility.
We explored the association of antibiotic-resistant phenotypes and genotypes in Acinetobacter spp with clinical outcomes and characteristics in 75 patients from a major military treatment facility. Amikacin resistance was associated with nosocomial acquisition of A baumannii, and carbapenem resistance and bla(OXA-23) were associated with the need for mechanical ventilation. The presence of bla(OXA-23) also correlated with longer hospital and ICU stay. Associations between bla(OXA-23) and complexity, duration, and changes made to antibiotic regimens also existed.


MicroRNA-34 Family and Its Role in Cardiovascular Disease.
MicroRNAs (miRNAs) have been considered important regulators in both physiological and disease contexts. Among all miRNAs, the miR-34 family (miR-34a, -34b, -34c), which has been well characterized as a tumor suppressor, displays diverse functions in noncancerous diseases. MiR-34 levels are relatively low in the cardiovascular system, but recently they have been reported to function in cardiovascular disorders by regulating apoptosis, telomere attrition, DNA damage, and inflammatory response. In this review, we mainly summarize their roles in cardiac senescence and ischemic diseases.


Micromorphological investigation of noncarious cervical lesions treated with demineralizing agents.
The purpose of this study was to examine the micromorphology of noncarious cervical lesions (NCCL) and the change occurring after acid conditioning. Nine wedge-shaped lesions, 9 saucer-shaped lesions, and 5 third molars with prepared cervical cavities were used. All NCCLs exhibited a hard, smooth surface with no discoloration. The teeth were sectioned and divided into three groups, each containing the three lesion types. Teeth in Group I were not etched, and teeth in Groups II and III were etched with 35% phosphoric acid and 20% polyacrylic acid/3% aluminum chloride, respectively. Field emission SEM and atomic force microscopy were used to examine the lesion surfaces. The surfaces of the prepared cervical (control) lesions were covered with a smear layer. Treatment with phosphoric acid and polyacrylic acid resulted in removal of the smear layer, although some residual smear layer was found on the surface treated with polyacrylic acid. The images of the untreated surfaces of NCCL showed highly mineralized dentin with complete obliteration of the dentinal tubules. After treatment with phosphoric and polyacrylic acids, the images revealed demineralization of the surface, which was more pronounced on those treated with phosphoric acid. The mineral deposits on the surface of NCCL decreased the effect of acid conditioning. Although there was no apparent difference between the two forms of NCCL used in this study, further work is needed to better understand these lesions.


Designing and evaluating an interprofessional education conference approach to antimicrobial education.
Arguably, Medical School curricula are deficient in learning opportunities related to the safe and effective use of medicines, in particular antimicrobials. Infection management is complex and multidisciplinary, and learning opportunities should reflect these principles. Aligned to the complexity of the subject matter, simulation and interprofessional based teaching are methods that can foster the collaborative skills required of future healthcare professionals. There have been calls to develop these methods in the teaching of safe prescribing and the management of infections; however, reports of such studies are limited. We developed an interprofessional education (IPE) conference for second year undergraduate medical and pharmacy students based in the North East of England. We considered contact theory in the design of three small group interprofessional workshops, on the broad themes of antimicrobial stewardship, infection management and patient safety. A mixed methods approach assessed students' attitudes towards IPE, barriers and facilitators of learning, and perceived learning gains. Qualitative data from workshop evaluation forms were analysed thematically, while quantitative data were analysed descriptively and differences between medical and pharmacy cohorts analysed using unpaired two-tailed t-tests. 226/352 students returned the workshop evaluation forms (66% of pharmacy students, 62% of medical students). 281/352 students responded to a series of Likert scale questions on the value of interprofessional education (88% of pharmacy students, 70% of medical students). Students reported acquisition of knowledge and skills, including concepts and procedures related to infection management and antimicrobial prescribing, and the development of problem-solving and critical evaluation skills. Students reflected on their attitude towards interprofessional collaboration. They reported a greater understanding of the roles of other healthcare professionals, reflected on the importance of effective communication in ensuring patient safety, and were more confident to work in interprofessional teams after the conference. A robust IPE event, theoretically underpinned by contact theory and developed collaboratively, achieved interprofessional learning at scale and helped develop healthcare professionals willing to collaborate across disciplines. The resources, and evaluation insights based on the 3P (presage, process, and product) model of learning and teaching, will be of value to other educators who seek to develop theoretically-sound IPE interventions.


Evolutionary dynamics of an ancient retrotransposon family provides insights into evolution of genome size in the genus Oryza.
Long terminal repeat (LTR) retrotransposons constitute a significant portion of most eukaryote genomes and can dramatically change genome size and organization. Although LTR retrotransposon content variation is well documented, the dynamics of genomic flux caused by their activity are poorly understood on an evolutionary time scale. This is primarily because of the lack of an experimental system composed of closely related species whose divergence times are within the limits of the ability to detect ancestrally related retrotransposons. The genus Oryza, with 24 species, ten genome types, different ploidy levels and over threefold genome size variation, constitutes an ideal experimental system to explore genus-level transposon dynamics. Here we present data on the discovery and characterization of an LTR retrotransposon family named RWG in the genus Oryza. Comparative analysis of transposon content (approximately 20 to 27,000 copies) and transpositional history of this family across the genus revealed a broad spectrum of independent and lineage-specific changes that have implications for the evolution of genome size and organization. In particular, we provide evidence that the basal GG genome of Oryza (O. granulata) has expanded by nearly 25% by a burst of the RWG lineage Gran3 subsequent to speciation. Finally we describe the recent evolutionary origin of Dasheng, a large retrotransposon derivative of the RWG family, specifically found in the A, B and C genome lineages of Oryza.