IL-10 regulates Il12b expression via histone deacetylation: implications for intestinal macrophage homeostasis.
To prevent excessive inflammatory responses to commensal microbes, intestinal macrophages, unlike their systemic counterparts, do not produce inflammatory cytokines in response to enteric bacteria. Consequently, loss of macrophage tolerance to the enteric microbiota plays a central role in the pathogenesis of inflammatory bowel diseases. Therefore, we examined whether the hyporesponsive phenotype of intestinal macrophages is programmed by prior exposure to the microbiota. IL-10, but not in vivo exposure to the microbiota, programs intestinal macrophage tolerance, because wild-type (WT) colonic macrophages from germ-free and specific pathogen-free (SPF)-derived mice produce IL-10, but not IL-12 p40, when activated with enteric bacteria. Basal and activated IL-10 expression is mediated through a MyD88-dependent pathway. Conversely, colonic macrophages from germ-free and SPF-derived colitis-prone Il10(-/-) mice demonstrated robust production of IL-12 p40. Next, mechanisms through which IL-10 inhibits Il12b expression were investigated. Although Il12b mRNA was transiently induced in LPS-activated WT bone marrow-derived macrophages (BMDMs), expression persisted in Il10(-/-) BMDMs. There were no differences in nucleosome remodeling, mRNA stability, NF-κB activation, or MAPK signaling to explain prolonged transcription of Il12b in Il10(-/-) BMDMs. However, acetylated histone H4 transiently associated with the Il12b promoter in WT BMDMs, whereas association of these factors was prolonged in Il10(-/-) BMDMs. Experiments using histone deacetylase (HDAC) inhibitors and HDAC3 short hairpin RNA indicate that HDAC3 is involved in histone deacetylation of the Il12b promoter by IL-10. These results suggest that histone deacetylation on the Il12b promoter by HDAC3 mediates homeostatic effects of IL-10 in macrophages.


Cervical lymphadenopathy following coronavirus disease 2019 vaccine: clinical characteristics and implications for head and neck cancer services.
Patients with coronavirus disease vaccine associated lymphadenopathy are increasingly being referred to healthcare services. This work is the first to report on the incidence, clinical course and imaging features of coronavirus disease vaccine associated cervical lymphadenopathy, with special emphasis on the implications for head and neck cancer services. This was a retrospective cohort study of all patients referred to our head and neck cancer clinics between 16 December 2020 and 12 March 2021. The main outcomes measured were the proportion of patients with vaccine-associated cervical lymphadenopathy, and the clinical and imaging characteristics. The incidence of vaccine-associated cervical lymphadenopathy referrals was 14.8 per cent (n = 13). Five patients (38.5 per cent) had abnormal-looking enlarged and rounded nodes with increased vascularity. Only seven patients (53.9 per cent) reported full resolution within an average of 3.1 ± 2.3 weeks. Coronavirus disease vaccine associated cervical lymphadenopathy can mimic malignant lymphadenopathy and therefore might prove challenging to diagnose and manage correctly. Healthcare services may encounter a significant increase in referrals.


Consultation and management of patients with lipid disorders: importance of life-style changes and intensive follow-up.
The purpose was to compare long-term involvement (LTI) vs short-term involvement (STI) of patients in a Lipid Education Service (LES) and to assess the effects of life-style changes. Patients were referred for LTI and STI by their physicians. Three-hundred eighty-one consecutive patients, 287 LTI and 94 STI, were analyzed. The lipids in LTI patients were all significantly improved. Sixty-six percent of all LDL values were at goal as were 77% of HDL and 62% of triglyceride (NCEP-II criteria). Sixty-two percent of LDL values in coronary artery disease patients were at goal contrasted to a reported range of 9.6%-71%. Life-style changes made significant differences in the percentage of patients at lipid goals. In postprogram follow-up there were significant differences between LTI and STI patients. The LTI patients did much better in improving lipid levels at postprogram follow-up than did STI patients. These results demonstrate the importance of making life-style changes as well as the importance of intensive efforts in achieving meaningful and lasting change.


Guidance document on delivery, treatment planning, and clinical implementation of IMRT: report of the IMRT Subcommittee of the AAPM Radiation Therapy Committee.
Intensity-modulated radiation therapy (IMRT) represents one of the most significant technical advances in radiation therapy since the advent of the medical linear accelerator. It allows the clinical implementation of highly conformal nonconvex dose distributions. This complex but promising treatment modality is rapidly proliferating in both academic and community practice settings. However, these advances do not come without a risk. IMRT is not just an add-on to the current radiation therapy process; it represents a new paradigm that requires the knowledge of multimodality imaging, setup uncertainties and internal organ motion, tumor control probabilities, normal tissue complication probabilities, three-dimensional (3-D) dose calculation and optimization, and dynamic beam delivery of nonuniform beam intensities. Therefore, the purpose of this report is to guide and assist the clinical medical physicist in developing and implementing a viable and safe IMRT program. The scope of the IMRT program is quite broad, encompassing multileaf-collimator-based IMRT delivery systems, goal-based inverse treatment planning, and clinical implementation of IMRT with patient-specific quality assurance. This report, while not prescribing specific procedures, provides the framework and guidance to allow clinical radiation oncology physicists to make judicious decisions in implementing a safe and efficient IMRT program in their clinics.


Nova Scotia hopes to solve recruitment problem with joint effort from MDS, government.
The Medical Society of Nova Scotia and the provincial government hope to solve the physician shortage in rural parts of the province with a recruiting effort that includes monetary incentives to fill some positions. A new locum service is designed to improve physician retention by making it easier to take vacations and pursue education programs.


A critical role for tetraspanin CD151 in alpha3beta1 and alpha6beta4 integrin-dependent tumor cell functions on laminin-5.
The basement membrane protein laminin-5 supports tumor cell adhesion and motility and is implicated at multiple steps of the metastatic cascade. Tetraspanin CD151 engages in lateral, cell surface complexes with both of the major laminin-5 receptors, integrins alpha3beta1 and alpha6beta4. To determine the role of CD151 in tumor cell responses to laminin-5, we used retroviral RNA interference to efficiently silence CD151 expression in epidermal carcinoma cells. Near total loss of CD151 had no effect on steady state cell surface expression of alpha3beta1, alpha6beta4, or other integrins with which CD151 associates. However, CD151-silenced carcinoma cells displayed markedly impaired motility on laminin-5, accompanied by unusually persistent lateral and trailing edge adhesive contacts. CD151 silencing disrupted alpha3beta1 integrin association with tetraspanin-enriched microdomains, reduced the bulk detergent extractability of alpha3beta1, and impaired alpha3beta1 internalization in cells migrating on laminin-5. Both alpha3beta1- and alpha6beta4-dependent cell adhesion to laminin-5 were also impaired in CD151-silenced cells. Reexpressing CD151 in CD151-silenced cells reversed the adhesion and motility defects. Finally, loss of CD151 also impaired migration but not adhesion on substrates other than laminin-5. These data show that CD151 plays a critical role in tumor cell responses to laminin-5 and reveal promotion of integrin recycling as a novel potential mechanism whereby CD151 regulates tumor cell migration.


A Teenager With Sickle Cell Disease and Scalp Swelling.
An intracranial bleed with a midline shift is a potentially life-threatening clinical condition. We present the unusual case of a 13-year-old boy with sickle cell disease who had numerous emergency department visits for a scalp hematoma and was subsequently determined to have subdural and epidural hematomas with midline shift, associated with a skull bone infarction. We review the pathophysiology of this unusual condition and emphasize the importance of including it in the differential diagnosis of any child with sickle cell anemia presenting with a nontraumatic scalp hematoma.


Two Antimicrobial Peptides Derived from Bacillus and Their Properties.
Growth promotion and disease prevention are important strategies in the modern husbandry industry, and for this reason, antibiotics are widely used as animal feed additives. However, the overuse of antibiotics has led to the serious problem of increasing resistance of pathogenic microorganisms, posing a major threat to the environment and human health. "Limiting antibiotics" and "Banning antibiotics" have become the inevitable trends in the development of the livestock feed industry, so the search for alternative antimicrobial agents has become a top priority. Antimicrobial peptides (AMPs) produced by Bacillus spp. have emerged as a promising alternative to antibiotics, due to their broad-spectrum antimicrobial activity against resistant pathogens. In this study, two strains of Bacillus velezensis 9-1 and B. inaquosorum 76-1 with good antibacterial activity were isolated from commercial feed additives, and the antimicrobial peptides produced by them were purified by ammonium sulfate precipitation, anion exchange chromatography, gel chromatography, and RP-HPLC. Finally, two small molecule peptides, named peptide-I and peptide-II, were obtained from strain 9-1 and 76-1, respectively. The molecular weight and sequences of the peptides were analyzed and identified by LC-MS/MS, which were 988.5706 Da and VFLENVLR, and 1286.6255 Da and FSGSGSGTAFTLR, respectively. The results of an antibacterial activity and stability study showed that the two peptides had good antibacterial activity against Staphylococcus aureus, B. cereus, and Salmonella enterica, and the minimum inhibitory concentrations were 64 μg/mL and 16 μg/mL, 32 μg/mL and 64 μg/mL, and 8 μg/mL and 8 μg/mL, respectively. All of them have good heat, acid, and alkali resistance and protease stability, and can be further developed as feed antibiotic substitutes.


Demonstration of mid-infrared waveguide photonic crystal cavities.
We have demonstrated what we believe to be the first waveguide photonic crystal cavity operating in the mid-infrared. The devices were fabricated from Ge23Sb7S70 chalcogenide glass (ChG) on CaF2 substrates by combing photolithographic patterning and focused ion beam milling. The waveguide-coupled cavities were characterized using a fiber end fire coupling method at 5.2 μm wavelength, and a loaded quality factor of ~2000 was measured near the critical coupling regime.


Infantile Alexander's disease. A case report.
The infantile Alexander disease is a leukodystrophy that appears in the early childhood, characterized by megaloencephaly, demyelination and presence of numerous Rosenthal fibers in the brain. This is an illustrative case in the study of patients with megaloencephaly and seizures, emphasizing the differential diagnosis. Our patient is a male infant aged 15 months with megaloencephaly, seizures, changes of behavior and delayed psychomotor development, with corroborated leukodystrophy demyelination in the brain, scanned by computed tomography (CT) and magnetic resonance (MR) imaging. It is a sporadic clinical case of infantile Alexander disease, without a known family history of the disorder. The final diagnosis has been confirmed by the magnetic resonance findings.


Increased Microglial Activity, Impaired Adult Hippocampal Neurogenesis, and Depressive-like Behavior in Microglial VPS35-Depleted Mice.
Vacuolar sorting protein 35 (VPS35) is a critical component of retromer, which is essential for selective endosome-to-Golgi retrieval of membrane proteins. VPS35 deficiency is implicated in neurodegenerative disease pathology, including Alzheimer's disease (AD). However, exactly how VPS35 loss promotes AD pathogenesis remains largely unclear. VPS35 is expressed in various types of cells in the brain, including neurons and microglia. Whereas neuronal VPS35 plays a critical role in preventing neurodegeneration, the role of microglial VPS35 is largely unknown. Here we provide evidence for microglial VPS35's function in preventing microglial activation and promoting adult hippocampal neurogenesis. VPS35 is expressed in microglia in various regions of the mouse brain, with a unique distribution pattern in a brain region-dependent manner. Conditional knocking out of VPS35 in microglia of male mice results in regionally increased microglial density and activity in the subgranular zone of the hippocampal dentate gyrus (DG), accompanied by elevated neural progenitor proliferation, but decreased neuronal differentiation. Additionally, newborn neurons in the mutant DG show impaired dendritic morphology and reduced dendritic spine density. When examining the behavioral phenotypes of these animals, microglial VPS3S-depleted mice display depression-like behavior and impairment in long-term recognition memory. At the cellular level, VPS35-depleted microglia have grossly enlarged vacuolar structures with increased phagocytic activity toward postsynaptic marker PSD95, which may underlie the loss of dendritic spines observed in the mutant DG. Together, these findings identify an important role of microglial VPS35 in suppressing microglial activation and promoting hippocampal neurogenesis, which are both processes involved in AD pathogenesis.SIGNIFICANCE STATEMENT The findings presented here provide the first in vivo evidence that Vacuolar sorting protein 35 (VPS35)/retromer is essential for regulating microglial function and that when microglial retromer mechanics are disrupted, the surrounding brain tissue can be affected in a neurodegenerative manner. These findings present a novel, microglial-specific role of VPS35 and raise multiple questions regarding the mechanisms underlying our observations. These findings also have myriad implications for the field of retromer research and the role of retromer dysfunction in neurodegenerative pathophysiology. Furthermore, they implicate a pivotal role of microglia in the regulation of adult hippocampal neurogenesis and the survival/integration of newborn neurons in the adult hippocampus.


Progressive telomere shortening occurs in cultured rat microglia, but not astrocytes.
Normal somatic cells have a finite replicative capacity. With each cell division, telomeres shorten progressively until they reach a critical length, at which point the cells enter replicative senescence. Some cells maintain their telomeres by the action of the telomerase enzyme. Glia, particularly microglia, are the only adult cell types in the central nervous system (CNS) that exhibit a significant mitotic potential, and are thus susceptible to telomere shortening. In this study, we show that telomere shortening accompanied by low to moderate telomerase activity, and ultimately senescence, occurs in rat microglia in vitro. When microglia are stimulated to divide with the mitogen granulocyte macrophage-colony stimulating factor (GM-CSF), longer telomeres are allowed to shorten, while shorter telomeres are lengthened. Telomerase activity is nearly 3-fold higher in GM-CSF-stimulated microglia initially, relative to unstimulated controls, and then declines to levels below those seen in controls before increasing again. Telomere attrition is also more rapid when microglia are grown in culture dishes of increasing size. Fluorescence in situ hybridization (FISH) analysis indicates that a nearly 3-fold variation in both inter- and intra-chromosomal telomere length exists in microglia. In contrast to microglia, cultured astrocytes exhibit a cyclical pattern of telomere lengthening and shortening over time, corresponding to a similar cycle of higher and lower telomerase activity. When astrocytes are passaged, mean telomere length increases initially from passage 1-2, remaining constant until passage 5, while the shortest telomeres are continually lengthened. In conclusion, the telomere shortening evident in microglia is accompanied by their progression to senescence by 32 days in vitro. In contrast, astrocytes, perhaps due to greater telomerase activity, have longer life spans and may be passaged repeatedly before entering senescence. Our findings provide an impetus to investigate the possibility that microglial telomere shortening may occur in vivo.


Kinetics of mushroom tyrosinase inhibition by quercetin.
The effects of quercetin on the activity of mushroom tyrosinase were studied. The equilibrium constants for this inhibitor binding with the enzyme molecule were established. The inhibition mechanism obtained from Lineweaver-Burk plots show that quercetin is a competitive inhibitor. In the time course of the oxidation of L-3,4-dihydroxyphenylalanine (L-DOPA) catalyzed by the enzyme in the presence of different concentrations of quercetin, the rate decreased with increasing time until a straight line was approached. The inhibition of tyrosinase by quercetin is a slow and reversible reaction with residual enzyme activity. The microscopic rate constants were determined for the reaction of quercetin with the enzyme.


Trans-ancestry genome-wide association meta-analysis of prostate cancer identifies new susceptibility loci and informs genetic risk prediction.
Prostate cancer is a highly heritable disease with large disparities in incidence rates across ancestry populations. We conducted a multiancestry meta-analysis of prostate cancer genome-wide association studies (107,247 cases and 127,006 controls) and identified 86 new genetic risk variants independently associated with prostate cancer risk, bringing the total to 269 known risk variants. The top genetic risk score (GRS) decile was associated with odds ratios that ranged from 5.06 (95% confidence interval (CI), 4.84-5.29) for men of European ancestry to 3.74 (95% CI, 3.36-4.17) for men of African ancestry. Men of African ancestry were estimated to have a mean GRS that was 2.18-times higher (95% CI, 2.14-2.22), and men of East Asian ancestry 0.73-times lower (95% CI, 0.71-0.76), than men of European ancestry. These findings support the role of germline variation contributing to population differences in prostate cancer risk, with the GRS offering an approach for personalized risk prediction.


Ultrastructural cytochemistry of atrial and ventricular cardiocytes of the bullfrog (Rana catesbeiana). Relationship of specific granules with reninlike activity of the myocardium.
The ultrastructural cytochemical reactivity, renin activity, and cathepsin D activity of atria and ventricle of the bullfrog have been assessed. The specific granules (A, B, and D) were found to be argentaphobic when ultrathin sections of Araldite-embedded atria and ventricle were stained according to the periodic acid-thiocarbohydrazide-silver proteinate technique of Thiery. The entire core of the specific granules was moderated positive after ultrathin sections of glutaraldehyde-fixed, glycol methacrylate embedded atria and ventricle were stained by phosphotungstic acid at a low pH. A similar reaction was shown by the cell coat, intercalated discs, residual bodies (C granules), and Z discs as well as by a very small portion of the Golgi complex. Incubation of ultrathin sections of atria and ventricule fixed only in glutaraldehyde and embedded in glycol methacrylate with either pronase or trypsin resulted in selective digestion of specific granules and Z discs and, to a much lesser degree, of the cell coat. As cathepsin D activity and renin activity were present in both atria and ventricle, the generation of angiotensin I by these cardiocytes might have been due to either enzyme. Nevertheless, because of the glycoprotein nature of specific granules and of the endocrinelike ultrastructure of atrial and ventricular cardiocytes in the frog, the present results raise the possibility that specific granules may contain renin.


Coronavirus Infection in Ferrets: Antigen Distribution and Inflammatory Response.
Multisystemic granulomatous lesions are the most common finding in ferrets infected by ferret systemic coronavirus (FRSCV). To characterize the inflammatory response developed against this virus, lesions from 4 naturally infected ferrets were examined. Lesions were classified into the 4 known types of granulomas (granulomas without necrosis [G], granulomas with necrosis [G-N], granulomas with neutrophils [G-NL], and diffuse granulomatous inflammation [DG]). The cellular composition of the lesions was characterized on the basis of cellular morphology and immunohistochemistry using markers for T and B-lymphocytes, plasma cells, macrophages, and neutrophils. The extent and distribution of viral antigen expression was also assessed. In G lesions, macrophages were mainly located in the center of the granuloma, with a moderate number of T-lymphocytes scattered among the macrophages, plasma cells, and B-lymphocytes. G-N lesions exhibited a necrotic center surrounded by abundant macrophages, some T-lymphocytes, plasma cells, and a few B-lymphocytes. In G-NL lesions, there was a central area dominated by neutrophils with low numbers of macrophages, plasma cells, and lymphocytes. DG presented similar cell proportions, but distributed evenly throughout the lesions. FRSCV was expressed in G, G-NL, G-N, and DG, with decreasing numbers of immunoreactive cells. This study reveals the important role of macrophages in the inflammatory response of ferrets against the virus and the variable proportions of leukocytes among different types of lesions, indicating their variable age. The results also confirm the similarities of the disease in ferrets to feline infectious peritonitis.


1,25-dihydroxyvitamin D3 increases the sensitivity of human renal carcinoma cells to tumor necrosis factor alpha but not to interferon alpha or lymphokine-activated killer cells.
Renal cell carcinoma is a chemotherapy-resistant tumor which is relatively responsive to immunotherapy. Immunotherapeutic regimes employ interferons or interleukin 2 with or without lymphokine-activated killer cells. Secondary cytokines, induced by interleukin 2 or interferon, may have an important impact on their anti-neoplastic activity. Notable among them is tumor necrosis factor (TNF alpha). We assessed the effect of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) on the susceptibility of the human renal cell carcinoma cell line SK-RC-29 to the cytotoxic and cytostatic actions of TNF alpha, interferon alpha and lymphokine-activated killer cells. Using uptake of the vital dye neutral red as an indicator of viable cell number, we found that addition of 1,25(OH)2D3 (100 nM) to TNF alpha (30 ng/ml)-treated cultures resulted in a 2.6 +/- 0.2-fold (mean +/- S.E.) increase in the cytotoxic effect of the cytokine. The potentiating effect of 1,25(OH)2D3 was dose-dependent, and significant at concentrations equal to or higher than 10 nM. Another dihydroxylated vitamin D metabolite, 24,25(OH)2D3, had no effect on TNF alpha action. The cytotoxic effect of TNF alpha increased whereas the potentiation by 1,25(OH)2D3 decreased with cell density in culture. 1,25(OH)2D3, in contrast to its potentiating effect on TNF alpha action, did not modulate the cytostatic effect of interferon alpha or the susceptibility of SK-RC-29 to killing by lymphokine-activated killer cells. The findings reported here may explain some of the in vivo anti-tumor activity of 1,25(OH)2D3 and provide a rationale for the employment of active vitamin D analogs during immune anti-cancer therapy.


Targeting the inhibition of fatty acid amide hydrolase ameliorate the endocannabinoid-mediated synaptic dysfunction in a valproic acid-induced rat model of Autism.
Autism spectrum disorder (ASD) is a neurodevelopmental disorder, characterized by social interaction impairment, stereotypical/repetitive behaviors and emotional deregulation. The endocannabinoid (eCB) system plays a crucial role in modulating the behavioral traits that are typically core symptoms of ASD. The major molecular mechanisms underlying eCB-dependent long-term depression (eCB-LTD) are mediated by group 1 metabotropic glutamate receptor (mGluR)-induced removal of postsynaptic α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPARs). Recently, modulation of anandamide (AEA), one of the main endocannabinoids in the brain, has been reported to alter social behaviors in genetic models of ASD. On this basis, we investigated the effects of treatment and the synaptic mechanism underlying AEA-mediated signaling in prenatal exposure to valproic acid (VPA) in rats. We found that the social deficits, repetitive behaviors and abnormal emotion-related behaviors in VPA-exposed offspring were improved after treatment with an inhibitor of AEA degrading enzyme, URB597. Using an integrative approach combing electrophysiological and cellular mechanisms, the results showed that the impaired eCB-LTD, abnormal mGluR-mediated LTD (mGluR-LTD) and decreased removal of AMPAR subunits GluA1 and GluA2 were reversed by URB597 in the prefrontal cortex (PFC) of VPA-exposed offspring. Taken together, these results provide the first evidence that rescue of the ASD-like phenotype by URB597 is mediated by enhancing the mechanism of removal of AMPAR subunits GluA1/2 underlying AEA signaling in the PFC in a VPA-induced model of ASD.


Impact of Y181C and/or H221Y mutation patterns of HIV-1 reverse transcriptase on phenotypic resistance to available non-nucleoside and nucleoside inhibitors in China.
The aim of this study was to investigate the role of K101Q, Y181C and H221Y emerging in HIV-1 reverse transcriptase with different mutations patterns in phenotypic susceptibility to currently available NNRTIs (nevirapine NVP, efavirenz EFV) and NRTIs (zidovudine AZT, lamivudine 3TC, stavudine d4T) in China. Phenotype testing of currently available NNRTIs (NVP, EFV) and NRTIs (AZT, 3TC, d4T) was performed on TZM-b1 cells using recombined virus strains. P ≤ 0.05 was defined significant considering the change of 50% inhibitory drug concentration (IC50) compared with the reference, while P ≤ 0.01 was considered to be statistically significant considering multiple comparisons. Triple-mutation K101Q/Y181C/H221Y and double-mutation K101Q/Y181C resulted in significant increase in NVP resistance (1253.9-fold and 986.4-fold), while only K101Q/Y181C/H221Y brought a 5.00-fold significant increase in EFV resistance. Remarkably, K101Q/H221Y was hypersusceptible to EFV (FC = 0.04), but was significantly resistant to the three NRTIs. Then, the interaction analysis suggested the interaction was not significant to NVP (F = 0.77, P = 0.4061) but significant to EFV and other three NRTIs. Copresence of mutations reported to be associated with NNRTIs confers significant increase to NVP resistance. Interestingly, some may increase the susceptibility to EFV. Certainly, the double mutation (K101Q/H221Y) also changes the susceptibility of viruses to NRTIs. Interaction between two different sites makes resistance more complex.


Six Hours of Manual Ventilation With a Bag-Valve-Mask Device Is Feasible and Clinically Consistent.
Manual ventilation of intubated patients is a common intervention. It requires skill as well as physical effort and is typically restricted to brief periods. Prolonged manual ventilation may be unavoidable in some scenarios, for example, extreme mass casualty incidents. The present study tested whether nurses are capable of appropriately manually ventilating patients for 6 hours. Volunteers performed ventilation on an electronic simulator for 6 hours while their own cardiorespiratory variables and the quality of the delivered ventilation were measured and recorded. The volunteers scored their perceived level of effort on a standard Borg Scale. Research laboratory at the Emergency Department, Tel Aviv Medical Center. Ten nursing staff members of the Tel Aviv Sourasky Medical Center, 25-43 years old. Volunteers ventilated manually a lung simulator for 6 hours. The subjects' physiologic states, including blood pressure, heart rate, respiratory rate, and oxygen saturation, showed no significant changes over time. The quality of delivered ventilation was somewhat variable, but it was stable on the average: average tidal volume ranged between 524.8 and 607.0 mL (p = 0.33). There was a slight but significant increase (7.3-10.9 L/min [p = 0.048]) in minute volume throughout the test period, reaching values consistent with mild hyperventilation. The subjects scored their perceived working effort as very light to fairly light, with a nonsignificant gradual increase in the Borg score as the study progressed. Manual ventilation of intubated patients can be performed continuously for 6 hours without excessive physical effort on the part of the operator. The quality of delivered ventilation was clinically adequate for all of them. There was a mild but significant trend toward hyperventilation, albeit within safe clinical levels, which was due to an increasing ventilatory rate rather than an increase in tidal volume.


Dynamic contrast-enhanced magnetic resonance angiography of the thoracic vessels: an intraindividual comparison of different k-space acquisition strategies.
The combination of parallel acquisition (generalized autocalibrating partially parallel acquisitions) and time-resolved three-dimensional (3D) view-sharing techniques is a promising tool for dynamic contrast-enhanced 3D-magnetic resonance angiography (MRA). We evaluated the influence of different k-space acquisition strategies on image quality for a recently developed time-resolved echo-shared angiographic technique during a contrast-enhanced 3D-MRA of the thoracic vessels. In 20 patients (16 men, 4 women; range, 28-75 years), 2 dynamic MRA protocols with different k-space acquisition strategies were performed on a 1.5-T whole-body scanner (MAGNETOM Avanto, Siemens AG, Erlangen, Germany) during injection of 5 mL (flow-rate, 3 mL/s) gadobutrol. For protocol 1, the central-region which was updated with every cycle included 20% of the entire k-space (protocol 2: 10%), the peripheral-region was undersampled by a factor of 10 (protocol 2: 5%). Image quality and details were compared visually. Signal-to-noise ratio and sharpness of vessel borders were estimated. Morphologic and functional assessment of the pulmonary arteries and the aorta was significantly improved for protocol 1. The sharpness of vessel borders (3.3 mm vs. 4.1 mm; P = 0.001), image quality, and the visibility of image details were significantly improved for protocol 1 compared with protocol 2. The size of the central region that is updated for every frame seems to be more crucial for image quality of echo-shared angiographic techniques than the sampling density in the periphery of the k-space.


Functional Testing or Coronary Computed Tomography Angiography in Patients With Stable Coronary Artery Disease.
The choice of either anatomical or functional noninvasive testing to evaluate suspected coronary artery disease might affect subsequent clinical management and outcomes. This study analyzed the association of initial noninvasive cardiac testing in outpatients with stable symptoms, with subsequent use of medications, invasive procedures, and clinical outcomes. We studied patients enrolled in a Danish nationwide register who underwent initial noninvasive cardiac testing with either coronary computed tomography angiography (CTA) or functional testing (exercise electrocardiography or nuclear stress testing) from 2009 to 2015. Further use of noninvasive testing, invasive procedures, medications, and medical costs within 120 days were evaluated. Risks of long-term mortality and myocardial infarction (MI) were analyzed using adjusted Cox proportional hazard models. A total of 86,705 patients underwent either functional testing (n = 53,744, mean age 57.4 years, 49% males) or coronary CTA (n = 32,961, mean age 57.4 years, 45% males), and were followed for a median of 3.6 years. Compared with functional testing, there was significantly higher use of statins (15.9% vs. 9.1%), aspirin (12.7% vs. 8.5%), invasive coronary angiography (14.7% vs. 10.1%), and percutaneous coronary intervention (3.8% vs. 2.1%); all p < 0.001 after coronary CTA. The mean costs of subsequent testing, invasive procedures, and medications were higher after coronary CTA ($995 vs. $718; p < 0.001). Unadjusted rates of mortality (2.1% vs. 4.0%) and MI hospitalization (0.8% vs. 1.5%) were lower after coronary CTA than functional testing (both p < 0.001). After adjustment, coronary CTA was associated with a comparable all-cause mortality (hazard ratio: 0.96; 95% confidence interval: 0.88 to 1.05), and a lower risk of MI (hazard ratio: 0.71; 95% confidence interval: 0.61 to 0.82). In stable patients undergoing initial evaluation for suspected coronary artery disease, coronary CTA was associated with greater use of statins, aspirin, and invasive procedures, and higher costs than functional testing. Coronary CTA was associated with a lower risk of MI, but a similar risk of all-cause mortality.


Metamorphosis in the Porosity of Recycled Concretes Through the Use of a Recycled Polyethylene Terephthalate (PET) Additive. Correlations between the Porous Network and Concrete Properties.
In the field of construction, sustainable building materials are currently undergoing a process of technological development. This study aims to contribute to understanding the behavior of the fundamental properties of concretes prepared with recycled coarse aggregates that incorporate a polyethylene terephthalate (PET)-based additive in their matrix (produced by synthesis and glycolysis of recycled PET bottles) in an attempt to reduce their high porosity. Techniques to measure the gas adsorption, water porosity, Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) were used to evaluate the effect of the additive on the physical, mechanical and microstructural properties of these concretes. Porosity reductions of up to 30.60% are achieved with the addition of 1%, 3%, 4%, 5%, 7% and 9% of the additive, defining a new state in the behavioral model of the additive (the overdosage point) in the concrete matrix; in addition, the porous network of these concretes and their correlation with other physical and mechanical properties are also explained.


Pepsinogen testing for evaluation of the success of Helicobacter pylori eradication at 4 weeks after completion of therapy.
Pepsinogen levels in plasma are increased by inflammation in the gastric mucosa, including inflammation resulting from Helicobacter pylori infection. A decrease in pepsinogen II level has been suggested as a reliable marker to confirm the successful eradication of infection. The aim of our study was to evaluate the potential role of pepsinogens I and II, gastrin-17 and H. pylori antibodies in confirming successful eradication. Altogether 42 patients (25 women, 17 men), mean age 45 years (range 23-74), were enrolled. Pepsinogens I and II, gastrin-17 and H. pylori IgG antibodies were measured in plasma samples using an ELISA test (Biohit, Oyj., Finland) before the eradication and 4 weeks after completing the treatment. The success of eradication was determined by a urea breath test. Eradication was successful in 31 patients (74%) and unsuccessful in 11 patients (26%). Pepsinogen II decreased significantly in both the successful (P=0.029) and unsuccessful (P=0.042) eradication groups. Pepsinogen I decreased significantly in the successful (P=0.025) but not the unsuccessful (P=0.29) eradication group. The pepsinogen I/II ratio increased in the successful eradication group (P=0.0018) but not in the group in which treatment failed (P=0.12). There were no differences in gastrin-17 or H. pylori antibody values. A decrease in pepsinogen II levels cannot be used as a reliable marker for the successful eradication of H. pylori 4 weeks after the completion of treatment. The increase in pepsinogen I/II ratio reflects differences in pepsinogen production following the eradication irrespective of improvement in atrophy.


NSP4 is stored in azurophil granules and released by activated neutrophils as active endoprotease with restricted specificity.
Whereas neutrophil elastase, cathepsin G, and proteinase 3 have been known as granule-associated serine proteases of neutrophils for decades, a fourth member, called neutrophil serine protease 4 (NSP4), was just recently described and provisionally characterized. In this study, we identified NSP4 as a novel azurophil granule protein of neutrophils by Western blot analyses of subcellular fractions as well as by RT-PCR analyses of neutrophil precursors from human bone marrow. The highest mRNA levels were observed in myeloblasts and promyelocytes, similar to myeloperoxidase, a marker of azurophil granules. To determine the extended sequence specificity of recombinant NSP4, we used an iterative fluorescence resonance energy transfer-based optimization strategy. In total, 142 different peptide substrates with arginine in P1 and variations at the P1', P2', P3, P4, and P2 positions were tested. This enabled us to construct an α1-proteinase inhibitor variant (Ile-Lys-Pro-Arg-/-Ser-Ile-Pro) with high specificity for NSP4. This tailor-made serpin was shown to form covalent complexes with all NSP4 of neutrophil lysates and supernatants of activated neutrophils, indicating that NSP4 is fully processed and stored as an already activated enzyme in azurophil granules. Moreover, cathepsin C was identified as the activator of NSP4 in vivo, as cathepsin C deficiency resulted in a complete absence of NSP4 in a Papillon-Lefèvre patient. Our in-depth analysis of NSP4 establishes this arginine-specific protease as a genuine member of preactivated serine proteases stored in azurophil granules of human neutrophils.


Theophylline and magnesium inhibit the contraction elicited with ciclosporin and angiotensin II in mesangial cell cultures.
One serious side effect of ciclosporin (CS) therapy is its acute nephrotoxicity characterized by a marked decrease in glomerular filtration rate (GFR). A main determinant of GFR is the ultrafiltration coefficient which is thought to be regulated by the contractile state of the cells of the mesangium. CS enhances contractions of mesangial cells elicited with angiotensin II. By way of a lowered ultrafiltration coefficient this effect of CS may be partly responsible for its acute nephrotoxicity. We have therefore examined the effect of compounds that may attenuate the enhanced contractile response of the smooth-muscle-like mesangial cells. Prostaglandin E2 (10(-8)-10(-5) mol/l) and dibutyryl cyclic AMP (10(-4) mol/l) reduced the number of contracting cells from more than 50% to about 10%. The clinically used agent theophylline (10(-5) g/ml) decreased the number of contractions of CS-pretreated mesangial cells from 80.0 to 28.4% (p less than 0.001). Additionally, a marked decrease in the percentage of contractions (83.3 to 33.3%; p less than 0.001) was observed when the concentration of magnesium chloride was elevated from 0 to 2 mmol/l. The finding that the enhanced contractile response of CS-pretreated mesangial cells can be overcome by theophylline and/or high magnesium levels may be of clinical interest with a view to acute CS nephrotoxicity.


Nitroglycerin tolerance and cyclic GMP generation in the longitudinal smooth muscle of the guinea-pig ileum.
The effects of in vitro nitroglycerin tolerance and methylene blue pretreatment on the ability of nitroglycerin and nitroprusside to promote relaxation and tissue accumulation of cyclic GMP were examined in the carbachol-contracted longitudinal smooth muscle of the guinea-pig ileum. Nitroglycerin and nitroprusside produced concentration-dependent increases in cyclic GMP levels. However, only nitroglycerin increased cyclic GMP levels before the onset of relaxation. Nitroglycerin tolerance produced approximately 200- and 5-fold shifts to the right of nitroglycerin and nitroprusside relaxation curves, respectively. Methylene blue pretreatment produced approximately 5-fold shifts to the right of both nitroglycerin and nitroprusside relaxation curves. Whenever there was an inhibition of nitroglycerin- or nitroprusside-induced relaxation, there was a corresponding reduction of cyclic GMP generation. Methylene blue inhibited the ability of 8-bromoguanosine 3',5'-monophosphoric acid to promote smooth muscle relaxation, suggesting that it also may impair the subsequent actions of cyclic GMP. This study provides the first demonstration of nitroglycerin tolerance in a nonvascular smooth muscle and provides evidence that cyclic GMP mediates the relaxant effects of nitroglycerin in the longitudinal smooth muscle of the guinea-pig ileum. However, because nitroprusside promoted tissue accumulation of cyclic GMP subsequent to relaxation, an exclusive role for cyclic GMP-mediated relaxation for this drug in the longitudinal smooth muscle appears unlikely.


NPY Y1 and Y5 receptor selective antagonists as anti-obesity drugs.
A combination of pharmacological and genetic studies in mice confirmed that the Y1 and Y5 receptors mediate the potent orexigenic actions of exogenous NPY. Although the physiological role of NPY in causing obesity is less clear, potent and selective antagonists of both Y1 and Y5 have been developed. Some of the NPY antagonists have suitable pharmacokinetic (PK) properties that allowed them to be evaluated in various rodent models of obesity. Several different Y1 and Y5 antagonists cause weight loss in rodent models, though confirmation that these effects are mechanism based has been limited. One Y5 antagonist, MK-0557 was evaluated in a 1-yr clinical trial and found to cause modest weight loss. Optimal NPY antagonist therapeutics for obesity may require blockade of both the Y1 and Y5 receptors.


Comprehensive home-based telerehabilitation in a morbidly obese male patient with severe obstructive sleep apnea. A case report.
Rehabilitation may be an effective additional treatment method in patients with obstructive sleep apnea (OSA). Physical exercise, weight reduction, pulmonary rehabilitation, and myofunctional therapy (MT) represent beneficial components of rehabilitation recommended as a possible adjunct to standard OSA treatment. A 54-year-old man with morbid obesity, long-lasting snoring, breathing pauses, frequent waking, as well as persistent drowsiness and fatigue during the day underwent polysomnography (PSG) to investigate suspected OSA. Severe OSA was confirmed by PSG and a 12-week comprehensive, home-based telerehabilitation program (tele-RHB program) along with continuous positive airway pressure (CPAP) therapy was implemented. The tele-RHB program included regular teleconsultations, aerobic-endurance training, MT, inspiratory and expiratory muscle training, as well as recommendations on proper nutrition, a healthy lifestyle, and behavioral changes. Following the treatment, the patient's quality of life (QoL), exercise capacity, lung function, and OSA severity significantly improved. The patient achieved an overall 19.9 kg reduction in weight, of which 16.2 kg was body fat, and his apnea-hypopnea index decreased by 42.6 episodes/hour. Our case report suggests that the comprehensive home-based tele-RHB program adjunct to CPAP therapy may be a novel approach for improving OSA severity, a patient's QoL, exercise capacity, lung function and body composition. It is important to note that such a program should be optional, however it may be needed to achieve the highest possible overall improvement in a patient's life. Further clinical investigations are needed to determine the therapeutic efficacy and clinical potential of this tele-RHB program.


Assessing Genetic Literacy Awareness and Knowledge Gaps in the US Population: Results from the Health Information National Trends Survey.
Public understanding of the role of genetics in disease risk is key to appropriate disease prevention and detection. This study assessed the current extent of awareness and use of genetic testing in the US population. Additionally, the study identified characteristics of subgroups more likely to be at risk for low genetic literacy. The study used data from the National Cancer Institute's 2017 Health Information National Trends Survey, including measures of genetic testing awareness, genetic testing applications and genetic testing usage. Multivariable logistic regression models estimated associations between sociodemographics, genetic testing awareness, and genetic testing use. Fifty-seven percent of respondents were aware of genetic tests. Testing awareness differed by age, household income, and race/ethnicity. Most participants had heard of using tests to determine personal disease risk (82.58%) or inherited disease risk in children (81.41%), but less were familiar with determining treatment (38.29%) or drug efficacy (40.76%). Among those with genetic testing awareness, actual testing uptake was low. A large portion of the general public lacks genetic testing awareness and may benefit from educational campaigns. As precision medicine expands, increasing public awareness about genetic testing applications for disease prevention and treatment will be important to support population health.


Mandibular lateral incisor-first premolar transposition in early mixed dentition: a rare case.
Dental transposition is an anomaly of eruption which is usually seen in the maxilla and rarely in the mandible. The mandibular lateral incisor's transposition with the permanent canine has a prevalence of less than 0.03%. The following case presents an even rarer situation, of a right lateral mandibular incisor that erupted in the place of the first premolar, in transposition with the temporary canine. In the first phase, a series of extractions have been performed, to ensure the necessary free space for moving the lateral incisor closer to its appropriate place, and after that a fixed orthodontic appliance was used. The biomechanics for intra-arch tooth movement was ingenious enough to ensure a perfect alignment of all permanent teeth and a functional Class I canine and molar final occlusion. Early intervention and successful move of the permanent canine in its correct position ensured the functionality of the mandibular movements and a good long-term prognosis.


The morphology of salt crystals affects the perception of saltiness.
High intake of salt (NaCl) has been associated with risk of non-communicable diseases, including hypertension, cardiovascular disease and stroke. Several strategies for reducing salt in foods are under study, including the relation of crystal morphology on dissolution properties of salt in the mouth. The aim of this paper was to study the dissolution of salt crystals with different morphologies in artificial saliva and to correlate the findings with the perception of saltiness over time. The morphology of five commercial salts was analyzed by scanning electronic microscopy and micro-CT studies. Shape parameters of crystals were determined using images from an optical microscope. Crystal dissolution in artificial saliva was evaluated using video-microscopy and the perception of saltiness was evaluated using sensorial test of time-intensity at standardized sodium content. Salt morphology was correlated well with dissolution rate and certain time-intensity parameters (time to maximum intensity, intensity at maximum and increase angle). Non-cubic and agglomerated crystals, such as Kosher and Maldon salts, were dissolved faster (dissolution rate up to 3.8 times higher) and experienced maximum saltiness (up to 17% more) at shorter times (up to 40% less). Crystal morphology may be a variable to consider to achieve sodium reduction while maintaining salt intensity.


Structural and physiological phenotypes of disease-linked lamin mutations in C. elegans.
The nuclear lamina is a major structural element of the nucleus and is predominately composed of the intermediate filament lamin proteins. Missense mutations in the human lamins A/C cause a family of laminopathic diseases, with no known mechanistic link between the position of the mutation and the resulting disease phenotypes. The Caenorhabditis elegans lamin (Ce-lamin) is structurally and functionally homologous to human lamins, and recent advances have allowed detailed structural analysis of Ce-lamin filaments both in vitro and in vivo. Here, we studied the effect of laminopathic mutations on Ce-lamin filament assembly in vitro and the corresponding physiological phenotypes in animals. We focused on three disease-linked mutations, Q159K, T164P, and L535P, which have previously been shown to affect lamin structure and nuclear localization. Mutations prevented the proper assembly of Ce-lamin into filament and/or paracrystalline arrays. Disease-like phenotypes were observed in strains expressing low levels of these mutant lamins, including decreased fertility and motility coincident with muscle lesions. In addition, the Q159K- and T164P-expressing strains showed a reduced lifespan. Thus, different disease-linked mutations in Ce-lamin exhibit major effects in vivo and in vitro. Using C. elegans as a model system, a comprehensive analysis of the effects of specific lamin mutations from the level of in vitro filament assembly to the physiology of the organism will help uncover the mechanistic differences between these different lamin mutations.


Development of quantitative polymerase chain reaction assays for allelic discrimination of gangliosidoses in cats.
To develop quantitative PCR (qPCR) assays with allele-specific primers to provide a rapid and accurate diagnostic and screening test for the 3 mutations identified as causes of gangliosidoses in domestic cats. DNA samples obtained from archived feline blood samples submitted for GM1 and GM2 testing. A qPCR assay was developed for each mutation to monitor the efficiency of PCR amplification. Results were determined on the basis of the fluorescent intensity of DNA staining. Samples from 60 cats were screened by use of the 3 qPCR assays. Of these, 59 qPCR results agreed with the sequence-derived genotypes. The phenotype (affected) for the other cat agreed with results for the qPCR assay, which indicated that interpretation of the sequence-based result was incorrect. The qPCR assays offer a sensitive, rapid, and reproducible technique for allelic discrimination without the need for complicated processing steps, such as hybridization or sequencing, after PCR procedures. These assays may prove beneficial for a rapid diagnosis of gangliosidoses in cats and could also provide a means for reliable large-scale screening for the carrier state, thereby accelerating the eradication of these debilitating diseases from feline populations.


Solar energy to biofuels.
In a solar economy, sustainably available biomass holds the potential to be an excellent nonfossil source of high energy density transportation fuel. However, if sustainably available biomass cannot supply the liquid fuel need for the entire transport sector, alternatives must be sought. This article reviews biomass to liquid fuel conversion processes that treat biomass primarily as a carbon source and boost liquid fuel production substantially by using supplementary energy that is recovered from solar energy at much higher efficiencies than the biomass itself. The need to develop technologies for an energy-efficient future sustainable transport sector infrastructure that will use different forms of energy, such as electricity, H(2), and heat, in a synergistic interaction with each other is emphasized. An enabling template for such a future transport infrastructure is presented. An advantage of the use of such a template is that it reduces the land area needed to propel an entire transport sector. Also, some solutions for the transition period that synergistically combine biomass with fossil fuels are briefly discussed.


Effects of short-term travel on COVID-19 spread: A novel SEIR model and case study in Minnesota.
The novel coronavirus responsible for COVID-19 was first identified in Hubei Province, China in December, 2019. Within a matter of months the virus had spread and become a global pandemic. In addition to international air travel, local travel (e.g. by passenger car) contributes to the geographic spread of COVID-19. We modify the common susceptible-exposed-infectious-removed (SEIR) virus spread model and investigate the extent to which short-term travel associated with driving influences the spread of the virus. We consider the case study of the US state of Minnesota, and calibrated the proposed model with travel and viral spread data. Using our modified SEIR model that considers local short-term travel, we are able to better explain the virus spread than using the long-term travel SEIR model. Short-term travel associated with driving is predicted to be a significant contributor to the historical and future spread of COVID-19. The calibrated model also predicts the proportion of infections that were detected. We find that if driving trips remain at current levels, a substantial increase in COVID-19 cases may be observed in Minnesota, while decreasing intrastate travel could help contain the virus spread.


Can simple clinical measurements detect patient noncompliance?
Measurement of patient compliance is essential if management of low compliance is to be performed efficiently. We assessed the value of several easily obtained clinical assessments compared to quantitative pill counts among 134 newly treated hypertensive male steelworkers during the first 6 months of their treatment with antihypertensive medication. Patient's self-reports obtained on structured interview correlated best with pill count compliance (r = 0.74, p less than 0.0001). Patients overestimated their compliance by an average of 17% but 90% of those who admitted to being noncompliant were found so. Qualitative urinary chlorthalidone and hydrochlorothiazide levels and changes in serum potassium, uric acid, and blood pressure also correlated with pill count compliance but were less accurate than interviews. Assessment of the patient's "health beliefs" and a variety of sociodemographic and health traits and perceptions did not provide useful information on compliance. Interviewing the patient is a simple and useful approach in assessing compliance with antihypertensive therapy.


A Novel Sparse Linear Mixed Model for Multi-Source Mixed-Frequency Data Fusion in Telemedicine.
Digital health and telemonitoring have resulted in a wealth of information to be collected to monitor, manage, and improve human health. The multi-source mixed-frequency health data overwhelm the modeling capacity of existing statistical and machine learning models, due to many challenging properties. Although predictive analytics for big health data plays an important role in telemonitoring, there is a lack of rigorous prediction model that can automatically predicts patients' health conditions, e.g., Disease Severity Indicators (DSIs), from multi-source mixed-frequency data. Sleep disorder is a prevalent cardiac syndrome that is characterized by abnormal respiratory patterns during sleep. Although wearable devices are available to administrate sleep studies at home, the manual scoring process to generate the DSI remains a bottleneck in automated monitoring and diagnosis of sleep disorder. To address the multi-fold challenges for precise prediction of the DSI from high-dimensional multi-source mixed-frequency data in sleep disorder, we propose a sparse linear mixed model that combines the modified Cholesky decomposition with group lasso penalties to enable joint group selection of fixed effects and random effects. A novel Expectation Maximization (EM) algorithm integrated with an efficient Majorization Maximization (MM) algorithm is developed for model estimation of the proposed sparse linear mixed model with group variable selection. The proposed method was applied to the SHHS data for telemonitoring and diagnosis of sleep disorder and found that a few significant feature groups that are consistent with prior medical studies on sleep disorder. The proposed method also outperformed a few benchmark methods with the highest prediction accuracy.


Cross validation for selection of cortical interaction models from scalp EEG or MEG.
A cross-validation (CV) method based on state-space framework is introduced for comparing the fidelity of different cortical interaction models to the measured scalp electroencephalogram (EEG) or magnetoencephalography (MEG) data being modeled. A state equation models the cortical interaction dynamics and an observation equation represents the scalp measurement of cortical activity and noise. The measured data are partitioned into training and test sets. The training set is used to estimate model parameters and the model quality is evaluated by computing test data innovations for the estimated model. Two CV metrics normalized mean square error and log-likelihood are estimated by averaging over different training/test partitions of the data. The effectiveness of this method of model selection is illustrated by comparing two linear modeling methods and two nonlinear modeling methods on simulated EEG data derived using both known dynamic systems and measured electrocorticography data from an epilepsy patient.


Pre- and intraoperative variables affecting early outcomes in elderly patients undergoing pancreaticoduodenectomy.
Conflicting data exist regarding the safety of pancreatic resections in elderly patients. In this study we compared early complication and mortality rates between patients younger and older than 80 years of age who underwent pancreaticoduodenectomy using a validated national database. The National Surgical Quality Improvement Program (NSQIP) database for 2005-2009 was used for this retrospective analysis. The primary outcome measures for our analysis were 30-day postoperative mortality, major complication rate and overall complication rate. A total of 6293 patients who underwent PD for any cause were included in the analysis. Of these, 9.4% were aged ≥80 years. The incidence of 30-day mortality was significantly higher in patients aged ≥80 years (6.3%) than in those aged <80 years (2.7%). Older patients were also noted to have higher rates of overall complications and serious complications. On multivariate analysis, age, ASA (American Society of Anesthesiologists) classification, reduced functional status, history of dyspnoea, and need for intraoperative transfusion were risk factors associated with the occurrence of overall complications, serious complications and postoperative mortality. This study shows that age among other factors is a determinant of postoperative morbidity and mortality following PD.


Evolutionary Dynamics of the OR Gene Repertoire in Teleost Fishes: Evidence of an Association with Changes in Olfactory Epithelium Shape.
Teleost fishes perceive their environment through a range of sensory modalities, among which olfaction often plays an important role. Richness of the olfactory repertoire depends on the diversity of receptors coded by homologous genes classified into four families: OR, TAAR, VR1, and VR2. Herein, we focus on the OR gene repertoire. While independent large contractions of the OR gene repertoire associated with ecological transitions have been found in mammals, little is known about the diversity of the OR gene repertoire and its evolution in teleost fishes, a group that includes more than 34,000 living species. We analyzed genomes of 163 species representing diversity in this large group. We found a large range of variation in the number of functional OR genes, from 15 in the Broad-nose Pipefish Syngnathus typhle and the Ocean Sunfish Mola mola, to 429 in the Zig-zag Eel Mastacembelus armatus. The number of OR genes was higher in species when a multilamellar olfactory rosette was present. Moreover, the number of lamellae was correlated with the richness of the OR gene repertoire. While a slow and balanced birth-and-death process generally drives the evolution of the OR gene repertoire, we inferred several episodes of high rates of gene loss, sometimes followed by large gains in the number of OR genes. These gains coincide with morphological changes of the olfactory organ and suggest a strong functional association between changes in the morphology and the evolution of the OR gene repertoire.


Genetic variability, structure and assignment of Spanish and French pig populations based on a large sampling.
The Spanish and French pig populations share the common practice of quasi systematic paternity control of pure breed and composite line males. Ten microsatellite markers are in common between Spain and France controls, among the 17 markers used in France and the 13 used in Spain. After the adjustment of allele sizes, it is possible to merge the two datasets and to obtain a set of 5791 animals, including the vast majority of the males in the Duroc, Landrace, Large White and Piétrain French and Spanish breeds. Twelve French composite lines are also available. The genetic diversity analysis of these pig populations is presented, as well as the assignment of an individual to its breed. The effects of heterogeneous sampling across time and of relatedness among animals are also assessed. Consistent with the results of the previous studies, we found that different populations from the same breed clearly clustered together. In addition, all populations of this study, whether purebred or composite, are quite well differentiated from the other ones. As a result, we note that the 10 microsatellites commonly used for paternity control ensure a powerful detection of the breed of origin, with the power of detection being 95-99%. The detection of the exact population within breed is more difficult, but the power exceeds 70% for most of the populations. Practical implications include, for instance, the detection of outlier animals, crosses and admixture events.


Enantioselective sensor based on microgravimetric quartz crystal microbalance with molecularly imprinted polymer film.
We report a novel quartz crystal microbalance sensor that provides enantioselectivity to dansylphenylalanine enantiomers by using a molecularly imprinted polymer film as a recognition element. The polymeric recognition thin film, imprinted with chiral dansyl-L-phenylalanine, was immobilised on a gold electrode modified with a photoactive precursor monolayer via a self-assembly process using photopolymerization. The fabricated sensor was able to discriminate between L- and D-dansylphenylalanine enantiomers in solution owing to the enantioselectivity of the imprinted sites. The enantiomeric composition of L- and D-enantiomeric mixtures could be quantitatively determined by the fabricated sensor. The detection limit is 5 micrograms mL-1 with a response range of 5-500 micrograms mL-1 at pH 10.0. The influence of the template concentration on the sensitivity and selectivity of the synthesised polymer membranes was investigated and optimised. The surface characteristics of the polymer coating were studied by varying the pH value of the buffer solution, and a convenient regeneration process was proposed to increase the reproducibility and reusability of the sensor by flushing with pH 2.0 buffer. The selectivity and recognition mechanism of the imprinted polymer film were studied with compounds that are structurally related to the template. The method presented in this work provides a novel means of preparing highly selective and sensitive chemical sensors via self-assembly and molecularly imprinting techniques.


Immunolocalization of the smooth muscle-specific protein calponin in complex and mixed tumors of the mammary gland of the dog: assessment of the morphogenetic role of the myoepithelium.
The immunohistochemical expression of the smooth muscle-specific protein calponin was studied to assess the contribution of myoepithelial cells to the histogenesis of spindle cells of complex and mixed tumors of the mammary gland of the dog and the origin of cartilage and bone in mixed tumors. Formalin-fixed tissues from 55 benign and malignant tumors (49 also containing surrounding normal mammary gland) were evaluated. Periacinar and periductal myoepithelial cells of all the 49 normal mammary glands were diffusely stained by the anti-human calponin monoclonal antibody. Calponin was found in 53 (98%) of the tumors studied, reacting with the myoepithelium-like cells of 86% of benign tumors and their remnants in 85% of malignant tumors. Five different types of calponin-immunoreactive myoepithelial cells were identified: hypertrophic myoepithelial cells. fusiform cells, stellate myoepithelial cells, rounded (myoepithelial) cells, and chondroblasts. Differences in staining intensity and staining pattern among these five types of cells suggested a transition of myoepithelial cells to chondroblasts. Stromal myofibroblasts also showed calponin immunoreactivity, but they did not react with a cytokeratin 14 monoclonal antibody, which recognizes myoepithelial cells in mammary gland. Calponin appears to be a very sensitive marker of normal and neoplastic myoepithelium in the canine mammary gland, and its identification in different cell types of complex and mixed tumors of the mammary gland of the dog suggests a major histogenetic role for myoepithelial cells.


Breast cancer as an acquired thrombophilic state.
Cancer is an acquired thrombophilic condition manifested by increased incidence of venous and arterial thromboembolic complications. Despite progress that has been achieved in treatments over the recent years, thromboembolism remains a major complication in patients with breast cancer; it is accompanied by significant morbidity and mortality. Approximately, 1% of breast cancer patients develop venous thromboembolism within 2 years with the highest incidence occurring in the 6 months post diagnosis. Metastatic disease and their comorbidities are the strongest predictors of the development of thrombotic event. The diagnosis of venous thromboembolism is associated with a higher risk of death within 2 years of diagnosis. Thromboembolic events in cancer patients range from abnormal laboratory coagulation tests without specific symptoms to massive thomboembolism and disseminated intravascular coagulation. The underlying pathophysiology is complex and includes the prothrombotic properties of cancer cells, which can be enhanced by anticancer treatment modalities, such as surgery, hormonal agents, and chemotherapy. Primary thromboprophylaxis in cancer patients should be individualized according to risk. For secondary prevention, several clinical studies have shown that low molecular weight heparin has improved patients' compliance, cancer outcomes and overall survival. This review summarizes the available data on the pathogenesis and clinical approach of hemostatic changes in breast cancer.


ECM modulated early kidney development in embryonic organ culture.
The use of exogenous signals is gaining importance in renal regenerative therapies. We wanted to explore the role of extracellular matrix (ECM) constituents on renal structure formation during renal organogenesis. We used a recently established organ culture setup to expose embryonic kidney rudiments directly to a large set of surface-immobilized or dissolved ECM molecules and growth factors. Organ culture was also performed on immobilized adult kidney ECM extracts and on reactive polymer films without any biomolecular components. The applied conditions resulted in distinct differences of organ phenotypes, underlining the multifaceted role of exogenous signals during kidney development. Specific ECM components, including collagen I and laminin, supported nephronal and tubular structure formation of the developing organ. ECM biopolymers, e.g. hyaluronic acid, were found to determine the fate of developing explants in a concentration- and molecular weight-dependent manner. The organ culture system used was an effective and robust means to identify exogenous signals that direct kidney development. This system can provide valuable insight for future regenerative therapies of kidney diseases.


Patient and Caregiver Preferences for First-Line Treatments of Metastatic Non-Small Cell Lung Cancer: A Discrete Choice Experiment.
The approval of immune checkpoint inhibitors for metastatic non-small-cell lung carcinomas (mNSCLC) treatment has presented more care options. Therefore, it is important to identify the benefit-risk trade-offs patients and caregivers are willing to make among potential treatment options. This study quantified the preferences of patients and caregivers for attributes of mNSCLC treatment. Patients with mNSCLC and caregivers completed an online survey assessing preferences using a discrete choice experiment. Respondents chose between hypothetical treatment profiles, with varying levels for 7 attributes associated with first-line treatment, including overall survival (OS), progression-free survival, select adverse events (AEs), and regimen (caregivers). Hierarchical Bayesian modeling was used to estimate attribute-level preference weights. Patients (n = 308) and caregivers (n = 166) most valued increasing OS from 11 to 30 months, followed by decreasing the risk of a serious AE (grade 3/4) that may lead to hospitalization from 70% to 18%. These attributes were over twice as important to both sets of respondents as the other attributes measured. Patients and caregivers would accept increases in the risks of a serious AE (grade 3/4) from 18% to 70% and all grades nausea from 10% to 69% if OS increased by 16.8 and 4.0 months, respectively. The least valued attributes were all grades of pneumonitis (patients) and all grades of skin rash (caregivers). Patients and caregivers are willing to make trade-offs between efficacy and toxicity and may require up to 1.5 years of increased OS to accept a higher risk of AEs. These results can provide guidance to oncologists when engaging in shared-decision making discussions.


A TGFβ-Smad4-Fgf6 signaling cascade controls myogenic differentiation and myoblast fusion during tongue development.
The tongue is a muscular organ and plays a crucial role in speech, deglutition and taste. Despite the important physiological functions of the tongue, little is known about the regulatory mechanisms of tongue muscle development. TGFβ family members play important roles in regulating myogenesis, but the functional significance of Smad-dependent TGFβ signaling in regulating tongue skeletal muscle development remains unclear. In this study, we have investigated Smad4-mediated TGFβ signaling in the development of occipital somite-derived myogenic progenitors during tongue morphogenesis through tissue-specific inactivation of Smad4 (using Myf5-Cre;Smad4(flox/flox) mice). During the initiation of tongue development, cranial neural crest (CNC) cells occupy the tongue buds before myogenic progenitors migrate into the tongue primordium, suggesting that CNC cells play an instructive role in guiding tongue muscle development. Moreover, ablation of Smad4 results in defects in myogenic terminal differentiation and myoblast fusion. Despite compromised muscle differentiation, tendon formation appears unaffected in the tongue of Myf5-Cre;Smad4(flox/flox) mice, suggesting that the differentiation and maintenance of CNC-derived tendon cells are independent of Smad4-mediated signaling in myogenic cells in the tongue. Furthermore, loss of Smad4 results in a significant reduction in expression of several members of the FGF family, including Fgf6 and Fgfr4. Exogenous Fgf6 partially rescues the tongue myoblast fusion defect of Myf5-Cre;Smad4(flox/flox) mice. Taken together, our study demonstrates that a TGFβ-Smad4-Fgf6 signaling cascade plays a crucial role in myogenic cell fate determination and lineage progression during tongue myogenesis.


Changes in triphasic mechanical properties of proteoglycan-depleted articular cartilage extracted from osmotic swelling behavior monitored using high-frequency ultrasound.
This study aims to obtain osmosis-induced swelling strains of normal and proteoglycan (PG) depleted articular cartilage using an ultrasound system and to investigate the changes in its mechanical properties due to the PG depletion using a layered triphasic model. The swelling strains of 20 cylindrical cartilage-bone samples collected from different bovine patellae were induced by decreasing the concentration of bath saline and monitored by the ultrasound system. The samples were subsequently digested by a trypsin solution for approximately 20 min to deplete proteoglycans, and the swelling behaviors of the digested samples were measured again. The bi-layered triphasic model proposed in our previous study (Wang et al., J Biomech Eng-Trans ASME 2007; 129: 413-422) was used to predict the layered aggregate modulus Ha from the data of depth-dependent swelling strain, fixed charge density and water content. It was found that the region near the bone, for the normal specimens, had a significantly higher aggregate modulus (Ha1 = 20.6 +/- 18.2 MPa) in comparison with the middle zone and the surface layer (Ha2 = 7.8 +/- 14.5 MPa and Ha3 = 3.6 +/- 3.2 MPa, respectively) (p < 0.001). The normalized thickness of the deep layer h1 was 0.68 +/- 0.20. After the trypsin digestion, the parametric values decreased to Ha1 = 13.6 +/- 9.6 MPa, Ha2 = 6.7 +/- 11.5 MPa, Ha3 = 2.7 +/- 3.2 MPa, and h1 = 0.57 +/- 0.28. Other models were also used to analyze data and the results were compared. This study showed that high-frequency ultrasound measurement combined with the triphasic modeling was capable of nondestructively quantifying the alterations in the layered mechanical properties of the proteoglycan-depleted articular cartilage.


Phylogeny and megasystematics of phagotrophic heterokonts (kingdom Chromista).
Heterokonts are evolutionarily important as the most nutritionally diverse eukaryote supergroup and the most species-rich branch of the eukaryotic kingdom Chromista. Ancestrally photosynthetic/phagotrophic algae (mixotrophs), they include several ecologically important purely heterotrophic lineages, all grossly understudied phylogenetically and of uncertain relationships. We sequenced 18S rRNA genes from 14 phagotrophic non-photosynthetic heterokonts and a probable Ochromonas, performed phylogenetic analysis of 210-430 Heterokonta, and revised higher classification of Heterokonta and its three phyla: the predominantly photosynthetic Ochrophyta; the non-photosynthetic Pseudofungi; and Bigyra (now comprising subphyla Opalozoa, Bicoecia, Sagenista). The deepest heterokont divergence is apparently between Bigyra, as revised here, and Ochrophyta/Pseudofungi. We found a third universal heterokont signature sequence, and deduce three independent losses of ciliary hairs, several of 1-2 cilia, 10 of photosynthesis, but perhaps only two plastid losses. In Ochrophyta, heterotrophic Oikomonas is sister to the photosynthetic Chrysamoeba, whilst the abundant freshwater predator Spumella is biphyletic; neither clade is specifically related to Paraphysomonas, indicating four losses of photosynthesis by chrysomonads. Sister to Chrysomonadea (Chrysophyceae) is Picophagea cl. nov. (Picophagus, Chlamydomyxa). The diatom-parasite Pirsonia belongs in Pseudofungi. Heliozoan-like actinophryids (e.g. Actinosphaerium) are Opalozoa, not related to pedinellids within Hypogyristea cl. nov. of Ochrophyta as once thought. The zooflagellate class Bicoecea (perhaps the ancestral phenotype of Bigyra) is unexpectedly diverse and a major focus of our study. We describe four new biciliate bicoecean genera and five new species: Nerada mexicana, Labromonas fenchelii (=Pseudobodo tremulans sensu Fenchel), Boroka karpovii (=P. tremulans sensu Karpov), Anoeca atlantica and Cafeteria mylnikovii; several cultures were previously misidentified as Pseudobodo tremulans. Nerada and the uniciliate Paramonas are related to Siluania and Adriamonas; this clade (Pseudodendromonadales emend.) is probably sister to Bicosoeca. Genetically diverse Caecitellus is probably related to Anoeca, Symbiomonas and Cafeteria (collectively Anoecales emend.). Boroka is sister to Pseudodendromonadales/Bicoecales/Anoecales. Placidiales are probably divergent bicoeceans (the GenBank Placidia sequence is a basidiomycete/heterokont chimaera). Two GenBank 'opalinid' sequences are fungal; Pseudopirsonia is cercozoan; two previous GenBank 'Caecitellus' sequences are Adriamonas.


Interplay of bromodomain and histone acetylation in the regulation of p300-dependent genes.
The bromodomain is an evolutionarily conserved motif found in many transcriptional activators including p300 which contains an intrinsic histone acetyltransferase activity and is a general coactivator for many transcription factors. One mode of bromodomain action is to serve as a binding module to recognize specific acetyl-lysine residue of histones during chromatin remodeling and transcriptional activation. The function of p300 is required for diverse sets of gene expression. However, it is not known whether the p300 bromodomain is involved in the expression of all or only subset of p300-dependent genes. In this study, we examined the impact of either wild type or a bromo-deficient p300 on the expression of several p300-dependant genes. The effects of histone acetylation on the expression of these genes were also assessed by targeting histone deacetylase activities with an inhibitor approach. We show that the impact of these inhibitors on the transcriptional activation of p300-dependent genes are impaired in cells containing the bromo-deficient p300, indicating that the interplay of p300 and histone acetylation in p300-dependent gene transcription requires the bromodomain. We also observed an increase in the expression of bromo-deficient p300 at the level of transcription possibly to compensate for the loss of p300 function. However, the high level of bromo-deficient p300 is not able to maintain the basal level of histone acetylation. Thus, the bromodomain is important for p300 to maintain the basal level of histone acetylation and to induce the transcriptional activation of p300-dependent genes. Nevertheless, the requirement of bromodomain and histone acetylation in p300-dependent gene transcription is determined by a gene specific manner.


Effect of a two-year supplementation with low doses of antioxidant vitamins and/or minerals in elderly subjects on levels of nutrients and antioxidant defense parameters.
Eighty-one elderly hospitalized subjects (> 65 years) were recruited for a double-blind placebo-controlled study to examine low dose supplementation of antioxidant vitamins and minerals on biological and functional parameters of free radical metabolism. Subjects were randomly assigned to one of the four treatment groups, daily receiving for 2 years: placebo group; mineral group: 20 mg zinc, 100 micrograms selenium; vitamin group: 120 mg vitamin C (Vit C), 6 mg beta-carotene (beta CA), 15 mg vitamin E (Vit E); mineral and vitamin group: Zn 20 mg, Se 100 micrograms, Vit C 120 mg, beta CA 6 mg, Vit E 15 mg. Fifty-seven subjects completed the study. A large frequency of Vit C, Zn and Se deficiencies were observed at baseline. As early as 6 months of treatment, a significant increase in vitamin and mineral serum levels was observed in the corresponding groups. The increases ranged from 1.1-4.0 fold depending on the nutrient. Antioxidant defense, studied in vitro with a test using red blood cells in presence of 2,2'-azo-bis (2-amidinopropane) by hydrochloride, showed an increase of cell resistance in patients receiving vitamins (p = 0.002); it was positively correlated with serum Vit C (p < 0.0001), alpha-tocopherol/cholesterol (p = 0.06), beta CA (p = 0.0014), serum Cu and Se (p < 0.05). Moreover, red blood cell antioxidant defense was reduced in elderly compared with young control subjects (50% hemolysis time: 69 +/- 14 mn and 109 +/- 12 mn, respectively). Erythrocyte glutathione peroxidase activity was enhanced in groups receiving minerals, whereas no significant change was observed for other indicators of oxidative stress (erythrocyte superoxide dismutase activity, thiobarbituric acid-reactive substances, total glutathione, reduced and oxidized forms). Our results provide experimental evidence that a low dose supplementation with vitamins and minerals was able to normalize biological nutrient status as early as 6 months of treatment. In addition, our data indicate that antioxidant defense in elderly subjects was improved with low doses of vit C, vit E and beta CA as studied by means of a functional test utilizing red blood cells challenged in vitro with free radicals.


The National Redress Scheme for Institutional Child Sexual Abuse - The Western Australian Response.
The Royal Commission into Institutional Responses to Child Sexual Abuse estimated 60,000 people were sexually abused as children in Australian Institutions during the period it examined and recommended the Australian Government establish a single National Redress Scheme. The National Redress for Institutional Child Sexual Abuse Act 2018(the NRS Act) commenced on 1 July 2018 enacting some, but not all, of the recommendations of the Royal Commission, including the implementation of the three elements of redress, the use of a matrix to determine monetary payments, and the provision of an internal review scheme. Redress under the NRS Act includes a "direct personal response by the institution", "counselling and psychological care" and "monetary payments". All States and Territories were required to respond to the NRS Act by referring relevant powers to the Commonwealth in order to facilitate the operation of the national scheme. In addition each jurisdiction enacted particular responses to the NRS having regard to the existing victims of crime legislation in each State or Territory. This article surveys the NRS Act and the approach adopted by Western Australia and the other sub-national jurisdictions in responding to the NRS Act.


Left ventricular wall movement in heart failure.
Of 34 patients admitted to hospital with left ventricular failure seven died before echocardiograms could be repeated after treatment and in three no echocardiograms could be obtained owing to chronic obstructive lung disease. In the remaining 24 patients echocardiograms were taken soon after admission and compared with echocardiograms taken later, after clinical improvement. The results show that in most patients both anterior and posterior motion of the posterior left ventricular wall increased. The rate of backward diastolic motion was appreciably less before and after treatment of heart failure compared with that in a small group of normal younger healthy men. This technique is a quick and apparently reliable way to assess ventricular function. The rate of diastolic motion is probably a reflection of left ventricular wall compliance.


Phase shifting interferometry: reference phase error reduction.
The accuracy of phase shifting interferometry is limited by reference phase errors caused by nonlinearities in the phase shifter movements, calibration errors of this device, phase drifts during the measuring interval, and low frequency mechanical vibrations. Since the error depends on the phase to be measured in a sinusoidal fashion one can greatly reduce its influence on the measuring results. For this purpose a function also containing the characteristic error function, is fitted to the measured data. In a second step the error function is subtracted from the measured phase values. In most cases an error reduction by 1 order of magnitude seems achievable as has been demonstrated by computer simulations.


Shape transitions in soft spheres regulated by elasticity.
We study elasticity-driven morphological transitions of soft spherical core-shell structures in which the core can be treated as an isotropic elastic continuum and the surface or shell as a tensionless liquid layer, whose elastic response is dominated by bending. To generate the transitions, we consider the case where the surface area of the liquid layer is increased for a fixed amount of interior elastic material. We find that generically there is a critical excess surface area at which the isotropic sphere becomes unstable to buckling. At this point it adopts a lower symmetry wrinkled structure that can be described by a spherical harmonic deformation. We study the dependence of the buckled sphere and critical excess area of the transition on the elastic parameters and size of the system. We also relate our results to recent experiments on the wrinkling of gel-filled vesicles as their interior volume is reduced. The theory may have broader applications to a variety of related structures from the macroscopic to the microscopic, including the wrinkling of dried peas, raisins, as well as the cell nucleus.


Electrocardiographic abnormalities and sudden death in myotonic dystrophy type 1.
Sudden death can occur as a consequence of cardiac-conduction abnormalities in the neuromuscular disease myotonic dystrophy type 1. The determinants of the risk of sudden death remain imprecise. We assessed whether the electrocardiogram (ECG) was useful in predicting sudden death in 406 adult patients with genetically confirmed myotonic dystrophy type 1. A patient was characterized as having a severe abnormality if the ECG had at least one of the following features: rhythm other than sinus, PR interval of 240 msec or more, QRS duration of 120 msec or more, or second-degree or third-degree atrioventricular block. Patients with severe abnormalities according to the entry ECG were older than patients without severe abnormalities, had more severe skeletal-muscle impairment, and were more likely to have heart failure, left ventricular systolic dysfunction, or atrial tachyarrhythmia. Such patients were more likely to receive a pacemaker or an implantable cardioverter-defibrillator during the follow-up period. During a mean follow-up period of 5.7 years, 81 patients died; there were 27 sudden deaths, 32 deaths from progressive neuromuscular respiratory failure, 5 nonsudden deaths from cardiac causes, and 17 deaths from other causes. Among the 17 patients who died suddenly in whom postcollapse rhythm was evaluated, a ventricular tachyarrhythmia was observed in 9. A severe ECG abnormality (relative risk, 3.30; 95% confidence interval [CI], 1.24 to 8.78) and a clinical diagnosis of atrial tachyarrhythmia (relative risk, 5.18; 95% CI, 2.28 to 11.77) were independent risk factors for sudden death. Patients with adult myotonic dystrophy type 1 are at high risk for arrhythmias and sudden death. A severe abnormality on the ECG and a diagnosis of an atrial tachyarrhythmia predict sudden death. (ClinicalTrials.gov number, NCT00622453.)


Robot-Assisted Vesico-Vaginal Fistula Repair: Our Technique and Review of the Literature.
Our first objective was to report our experience on robot-assisted vesico-vaginal fistula (VVF) repair after radical surgery for gynecologic malignancies without omental flap interposition using a da Vinci robotic system. The second objective was to critically review the literature in order to analyze surgical solutions used to avoid repair failure. Five patients with VVF diagnosed after previous open surgery for gynecologic malignancies referred to our tertiary institutions were selected. After an adequate oncologic follow-up, based on the fistula diameter and conservative management failure, robotic surgery repair was proposed. A bivalve 2-layer suturing technique was carried out without tissue interpositions; omentum was not available secondary to previous surgery including hysterectomy, ovaryectomy, and omentectomy and citoreductive peritoneomectomy. A systematic review of the literature was performed in December 2016 using the PubMed database with the following keywords: robotic, robot-assisted, vesico-vaginal, fistula repair. Median age was 62 years (range 55-71) bearing long-lasting VVF were referred to our divisions. Median fistula diameter was 5 mm (range 3-8 mm). Fistula site was the trigone and identified during cystoscopy near the mid-line, left, and right urether meatus, respectively. The median overall and console operatory time were 250 and 120 min, respectively. Blood loss was insignificant (median 40 mL) and the median length of stay was 7 days without any complication. Ten papers were found fulfilling the mentioned criteria, from which 6 were case reports, single or multiple, accounting for the overall 41 robotic-approach-operated patients. The quality of the dissection and suture associated with efficient urine drainage are in our opinion the key elements of the success of our technique, which can be performed even without omentum or other tissue flap or graft interposition.


A Natural Triterpenoid Saponin as Multifunctional Stabilizer for Drug Nanosuspension Powder.
The objective of this study is to prepare a novel drug nanosuspension modified by a natural triterpenoid saponin (glycyrrhizin (GZ)) and evaluate its stability and redispersibility. A poorly soluble drug (andrographolide (AGE)) was used as a model drug. AGE nanosuspensions (AGE-NS) using GZ as natural stabilizer with mean particle size of 487 nm were firstly prepared by homogenization and converted into dried AGE nanosuspension powder (AGE-NP) by freeze-drying. It was found that GZ was able to prevent the aggregation of AGE nanocrystals and the freeze-dried AGE-NP could easily redisperse back to AGE-NS. It was related with special properties of GZ that possessed the interfacial property (37.02 ± 0.29 N/m) and electrostatic effect (-43.6 ± 0.9 mV) and could entrap AGE nanocrystals into its network structure. The freeze-dried AGE-NP/GZ exhibited excellent performance, compared with those combined with trehalose as matrix formers. The powder X-ray diffraction result demonstrated that GZ did not alter the AGE crystal state. The dissolution of AGE-NP/GZ (99.87%) was significantly enhanced, compared with the coarse AGE (42.35%). This study demonstrated that GZ could be used as a novel multifunctional stabilizer for production of drug nanosuspensions and provided a promising basis for further formulation development of poorly soluble drug.


Hierarchy and roles of pathogen-associated molecular pattern-induced responses in Nicotiana benthamiana.
Our current understanding of pathogen-associated molecular pattern (PAMP)-triggered immunity signaling pathways in plants is limited due to the redundancy of several components or the lethality of mutants in Arabidopsis (Arabidopsis thaliana). To overcome this, we used a virus-induced gene silencing-based approach in combination with pharmacological studies to decipher links between early PAMP-triggered immunity events and their roles in immunity following PAMP perception in Nicotiana benthamiana. Two different calcium influx inhibitors suppressed the reactive oxygen species (ROS) burst: activation of the mitogen-activated protein kinases (MAPKs) and PAMP-induced gene expression. The calcium burst was unaffected in plants specifically silenced for components involved in ROS generation or for MAPKs activated by PAMP treatment. Importantly, the ROS burst still occurred in plants silenced for the two major defense-associated MAPK genes NbSIPK (for salicylic acid-induced protein kinase) and NbWIPK (for wound-induced protein kinase) or for both genes simultaneously, demonstrating that these MAPKs are dispensable for ROS production. We further show that NbSIPK silencing is sufficient to prevent PAMP-induced gene expression but that both MAPKs are required for bacterial immunity against two virulent strains of Pseudomonas syringae and their respective nonpathogenic mutants. These results suggest that the PAMP-triggered calcium burst is upstream of separate signaling branches, one leading to MAPK activation and then gene expression and the other to ROS production. In addition, this study highlights the essential roles of NbSIPK and NbWIPK in antibacterial immunity. Unexpectedly, negative regulatory mechanisms controlling the intensity of the PAMP-triggered calcium and ROS bursts were also revealed by this work.


Impact of lipoatrophy on patient-reported outcomes in antiretroviral-experienced patients.
Lipoatrophy is a noteworthy adverse effect of antiretroviral therapy. A 2-part literature review was conducted to assess the impact of lipoatrophy in HIV-infected persons: the first reviewed the qualitative studies that reported lipoatrophy data, and the second reviewed the clinical studies that recorded patient-reported outcome end points. This literature review provided considerable evidence that lipoatrophy has a negative impact on the psychosocial well-being and health-related quality of life of HIV-infected persons receiving antiretroviral therapy. However, the patient-reported outcome measures used in clinical studies were not lipoatrophy-specific and had limited ability to show real changes in health status associated with lipoatrophy.


Rehabilitation of the painful upper extremity.
Therapy for the painful upper extremity is a challenge. This article focuses on various assessment tools and techniques that are reflective of the painful status. The author reviews several research articles that discuss validating treatment options and outlines rehabilitation for the painful upper extremity as it relates to peripheral nerve irritation and compression.


Modulating effects of thyme and its major ingredients on oxidative DNA damage in human lymphocytes.
The present study was carried out to investigate the modulating effects of thyme and its major components against the oxidative DNA damage induced by H(2)O(2). The human lymphocytes with thymol, carvacrol, and gamma-terpinene incubated with or without 0.1 mM H(2)O(2) for 30 min at 37 degrees C and the DNA damage were evaluated by singe cell gel electropheresis (comet assay). Concentrations above 0.1 mM thymol and gamma-terpinene and 0.05 mM carvacrol significantly induced DNA damage in human lymphocytes, but at the smaller concentrations no additional DNA strand breakage has been observed. At the all concentrations studied, gamma-terpinene did not show any protective effect against H(2)O(2) induced oxidative DNA damage, but the phenolic compounds thymol and carvacrol at concentrations below 0.2 and 0.1 mM, respectively, significantly reduced the oxidative DNA damage (p < 0.001). The n-hexane and ethyl acetate fractions prepared from the methanolic extracts of Thymus spicata also were found to inhibit DNA damage.


Determining divergence times with a protein clock: update and reevaluation.
A recent study of the divergence times of the major groups of organisms as gauged by amino acid sequence comparison has been expanded and the data have been reanalyzed with a distance measure that corrects for both constraints on amino acid interchange and variation in substitution rate at different sites. Beyond that, the availability of complete genome sequences for several eubacteria and an archaebacterium has had a great impact on the interpretation of certain aspects of the data. Thus, the majority of the archaebacterial sequences are not consistent with currently accepted views of the Tree of Life which cluster the archaebacteria with eukaryotes. Instead, they are either outliers or mixed in with eubacterial orthologs. The simplest resolution of the problem is to postulate that many of these sequences were carried into eukaryotes by early eubacterial endosymbionts about 2 billion years ago, only very shortly after or even coincident with the divergence of eukaryotes and archaebacteria. The strong resemblances of these same enzymes among the major eubacterial groups suggest that the cyanobacteria and Gram-positive and Gram-negative eubacteria also diverged at about this same time, whereas the much greater differences between archaebacterial and eubacterial sequences indicate these two groups may have diverged between 3 and 4 billion years ago.


[Deguelin regulates cell cycle and nuclear pore complex protein Nup98 and Nup88 in U937 cells in vitro].
To investigate antitumor activity and molecular mechanism of deguelin to the human U937 leukaemia cells and to explore the mechanisms regulating cell cycle and nucleoporin 98 (Nup98) and nucleoporin 88 (Nup88) in vitro. The effects of deguelin on the growth of U937 cells were studied by MTT assay, and the cell cycle of U937 cells by a propidium iodide method. The localization of the nuclear pore complex protein Nup98 and Nup88 was checked by immunofluorescence and immunoelectron microscopy. The expressions of Nup98 and Nup88 in U937 cells were checked by flow cytometry (FCM) and Western blot respectively. The proliferation of U937 cells was significantly inhibited in a time-dose dependent manner in deguelin-treated group with a 24 h IC50 value of 21.61 nmol/L and 36 h IC50 value of 17.07 nmol/L. U937 cells treated with deguelin showed reduction in the percentages of cells in G0/G1, whereas accumulation of cells in S and G2/M phase. The ratio of G1/G0 phase cells were 73.01%, 71.15%, 68.42%, 52.45%, 43.99% and 22.82%, and that of S phase cells were 17.18%, 16.30%, 18.09%, 27.56%, 31.21% and 46.85%, and that of G2/M phase cells were 9.75%, 12.31%, 13.09%, 18.99%, 24.83% and 27.79% at deguelin concentrations of 0, 5, 10, 20, 40, 80 nmol/L respectively. Nup88 and Nup98 were found on both the nuclear and cytoplasmic side of the U937 cells. The expression of Nup98 was up-regulated and Nup88 down-regulated in deguelin treated U937 cells. Deguelin is able to inhibit the proliferation of U937 cells by regulating the cell cycle. The antitumor activity of deguelin was related to up-regulating the expression of Nup98 and down-regulating Nup88 protein.