Harnessing the chemical activation inherent to carrier protein-bound thioesters for the characterization of lipopeptide fatty acid tailoring enzymes.
Here, we report a new experimental approach utilizing an amide ligation reaction for the characterization of acyl carrier protein (ACP)-bound reaction intermediates, which are otherwise difficult to analyze by traditional biochemical methods. To explore fatty acid tailoring enzymes of the calcium-dependent antibiotic (CDA) biosynthetic pathway, this strategy enabled the transformation of modified fatty acids, covalently bound as thioesters to an ACP, into amide ligation products that can be directly analyzed and compared to synthetic standards by HPLC-MS. The driving force of the amide formation is the thermodynamic activation inherent to thioester-bound compounds. Using this novel method, we were able to characterize the ACP-mediated biosynthesis of the unique 2,3-epoxyhexanoyl moiety of CDA, revealing a new type of FAD-dependent oxidase HxcO with intrinsic enoyl-ACP epoxidase activity, as well as a second enoyl-ACP epoxidase, HcmO. In general, our approach should be widely applicable for the in vitro characterization of other biosynthetic systems acting on carrier proteins, such as integrated enzymes from NRPS and PKS assembly lines or tailoring enzymes of fatty and amino acid precursor synthesis.


[Complex treatment of patients with destructive pulmonary tuberculosis including intramuscular administration of isoniazid].
The results of a complex treatment of 126 patients with destructive pulmonary tuberculosis have been studied. The chemotherapy regimen of 72 patients comprised intramuscular administration of a 10% isoniazid solution and 54 received isoniazid orally. Both groups of patients were identical in the clinical pattern of pulmonary tuberculosis and treatment conditions. Treatment effectiveness proved to be higher in Group I patients in whom the hospital stay was mainly 6 months. With oral isoniazid, it was longer and 24% of patients (versus 9.7% in Group I) required more prolonged therapy. Clinical manifestations, which were identical in both groups in the pre-treatment period, subsided earlier when isoniazid was administered intramuscularly. Hence, intramuscular administration of 10% isoniazid solution, as compared with its oral use, raises the effectiveness of chemotherapy and decreases the terms of a hospital treatment.


Autonomic control mechanism of maximal lower body negative pressure application.
Autonomic control mechanisms during progressive hemorrhage in humans remain complex and unclear. The present study investigates the autonomic reflexes during maximal application of lower body negative pressure (LBNP) that mimics severe hemorrhage in conscious human subjects (n=10) using analyses of heart rate variability (HRV) and systolic blood pressure variability (BPV) and baroreflex sensitivity. Spectral analysis of HRV included linear power spectral density (PSD), and nonlinear principal dynamic modes (PDM) methods. The maximal LBNP application decreased (P<0.01) the systolic and pulse pressures (PP), root mean square successive differences, normalized high frequency (HF) power of HRV, and transfer function gains at low frequency (LF) and HF bands. Meanwhile, increases (P<0.05) in heart rate, diastolic blood pressure (DBP), LFHRV, LF/HFHRV, and sympathetic activity of HRV using PDM were observed during maximal LBNP tolerance. After the termination of LBNP, no significant changes (P>0.05) were found in all the parameters except DBP and PP between recovery and baseline conditions. Rapid application of maximal LBNP that simulated severe hemorrhage was found to be associated with unloading of baroreflex mediated increased sympathetic reflex.


Structural and electronic properties of oligo- and polythiophenes modified by substituents.
The electronic and structural properties of oligo- and polythiophenes that can be used as building blocks for molecular electronic devices have been studied by using periodic density functional theory calculations. We have in particular focused on the effect of substituents on the electronic structure of thiophenes. Whereas singly bonded substituents, such as methyl, amino or nitro groups, change the electronic properties of thiophene monomers and dimers, they hardly influence the band gap of polythiophene. In contrast, phenyl-substituted polythiophenes as well as vinyl-bridged polythiophene derivatives exhibit drastically modified band gaps. These effects cannot be explained by simple electron removal or addition, as calculations for charged polythiophenes demonstrate.


Mesenchyme-mediated effects of retinoic acid during rat intestinal development.
In previous experiments we showed that intestinal development was dependent upon epithelial-mesenchymal cell interactions. The aim of this study was to investigate the possible role of retinoic acid (RA), a morphogenetic and differentiating agent, on the gut epithelial-mesenchymal unit. For this purpose we first analyzed the effects of a physiological dose of RA on 14-day fetal rat intestine using short-term organ culture experiments, or long-term grafts under the skin of nude mice. In these conditions, RA accelerated villus outgrowth and epithelial cell differentiation as assessed by the onset of lactase expression, and it also stimulated muscle and crypt formation. In order to analyze potential effects of RA mediated by mesenchymal cells, we isolated and characterized gut mucosa mesenchyme-derived cell cultures (mesenchyme-derived intestinal cell lines, MIC). These cells were shown to express mRNAs for retinoid binding proteins similar to those expressed in situ in the intestinal mesenchyme. MIC cells co-cultured with 14-day intestinal endoderms promoted endodermal cell adhesion and growth, and the addition of exogeneous RA enhanced epithelial cell polarization and differentiation assessed by cytokeratin and lactase immunostaining. Such a differentiating effect of RA was not observed on endodermal cells when cultured without a mesenchymal feeder layer or maintained in conditioned medium from RA-treated MIC cells. In the co-cultures, immunostaining of laminin and collagen IV with polyclonal antibodies, as well as alpha1 and beta1 laminin chains mRNAs (analyzed by RT-PCR) increased concurrently with the RA-enhanced differentiation of epithelial cells. It is worth noting that this stimulation by RA was also obvious on the mesenchymal cells cultured alone. These results show that RA plays a role in intestinal morphogenesis and differentiation. In addition, they indicate that RA acts on the mesenchymal cell phenotype and suggest that RA may modify the mesenchymal-epithelial cell interactions during intestinal development.


Sonography of the iliopsoas tendon and injection of the iliopsoas bursa for diagnosis and management of the painful snapping hip.
The purpose of this study was to compare sonographic evaluations of patients referred with suspected snapping of their iliopsoas tendon with the pain relief achieved from anesthetic injection of the iliopsoas bursa, and with the subsequent surgical outcome. This study also assessed the effectiveness of Kenalog injection into the iliopsoas bursa for long-term pain relief. Dynamic and static sonography was performed in 40 patients with clinically diagnosed snapping hips. The iliopsoas bursa was injected with Bupivicaine and Lidocaine in the first 22 patients, and an additional 1 ml Kenalog-40 was added to this mixture in the last 18 patients. We compared the static and dynamic sonographic findings with change in the patients' level of pain at 2 days after anesthetic injection. The sonographic findings and response to anesthetic injection were also compared to the response to Kenalog injection and the results of any subsequent surgery. Static sonography of the iliopsoas tendon was normal in 38 patients, and detected iliopsoas bursitis in one patient and iliopsoas tendinopathy in another. Snapping of the iliopsoas tendon was observed using dynamic sonography in 9 of the 40 patients. Following anesthetic injection of the iliopsoas bursa, 29 patients had complete or partial pain relief, and 11 patients had no pain relief. Eight of the nine patients with a snapping iliopsoas tendon had complete or partial pain relief from the bursal injection. Twelve of the 29 patients with pain relief after anesthetic injection later had an arthroscopic iliopsoas tendon release, and all of these 12 patients had a good postoperative result. Of the 18 patients who had Kenalog-40 injected into the iliopsoas bursa and did not have iliopsoas surgery, 16 had sustained pain relief following the injection. Patients with groin pain and a clinically suspected snapping iliopsoas tendon can benefit from injection into the iliopsoas bursa even if the snapping tendon is not visualized sonographically. The use of a corticosteroid may provide long-term pain relief, and pain relief after injection is a predictor of good outcome after surgical release of the iliopsoas tendon.


Endogenous nitric oxide production in canine osteoarthritis: Detection in urine, serum, and synovial fluid specimens.
To measure nitric oxide (NO) concentrations in serum, urine, and synovial fluid (SF) of dogs with naturally occurring cranial cruciate ligament (CCL) rupture and normal dogs, and to compare these with clinical and histologic changes of osteoarthritis (OA). Prospective clinical study including 2 groups of animals selected from the hospital population. Forty-three dogs (CCL group) with OA secondary to CCL rupture; 30 healthy dogs (control group) without CCL rupture. Serum, urine, and SF were collected before and during surgery in the CCL group or immediately after euthanasia in the control group. Articular cartilage and synovial membrane tissue specimens were prepared for routine histologic examination. The stable end products of NO, total nitrite and nitrate (NOt) activity, were measured in body fluids and compared with macroscopic and histologic degrees of OA. Urinary NOt concentration was compared with urinary creatinine concentration and stated as urinary NOt:creatinine ratio (UNCR). RESULTS-SF NOt concentrations were not significantly different between the 2 groups. Serum NOt concentrations (45.6 vs 28.9 micromol/L; P =.042) and the UNCR (0.007 vs 0.004; P =.035) were significantly higher in dogs of the CCL group compared with the control population. An association between UNCR and histologic and macroscopical OA grades could be demonstrated. UNCR might be a useful indicator of nitrite and nitrate production and, therefore, osteoarthritic changes in joints. UNCR could be used as a tool to evaluate the NOt production by joint tissues over time and might therefore provide a method of evaluating the effects of drugs in the control of osteoarthritis.


Ecological specialization in Diaeretiella rapae (Hymenoptera: Braconidae: Aphidiinae) on aphid species from wild and cultivated plants.
Diaeretiella rapae is an aphid parasitoid with potential for use in biological control strategies. However, several recent genetic studies have challenged the long held view that it is a generalist parasitoid. We investigated its ecological specialization and ability to use resources in cultivated and uncultivated areas. Ecological specialization would reduce its ability to exploit the diversity of aphid species, particularly in uncultivated areas, and to control pest aphids. Four D. rapae strains were studied, three reared on pest aphids on Brassicaceae and one strain on a non-pest aphid on Chenopodiaceae. For each strain, we performed host-switching experiments, with a total of six aphid species, five of which D. rapae parasitizes in France. We tested cross-breeding ability between strains to detect potential reproductive isolation linked to aphid host species in D. rapae. The strain reared on non-pest aphids was able to develop on aphid species from both cultivated and uncultivated plants. The strains reared on pest aphids, however, exclusively parasitized aphid species on cultivated Brassicaceae. In addition, reproductive isolation was detected between strains from uncultivated and cultivated plants. Thus, the D. rapae populations examined here appear to be showing ecological specialization or they may even be composed of a complex of cryptic species related to the aphid hosts. The role of Chenopodium album as a reservoir for D. rapae, by providing a habitat for non-pest aphids on which it can feed, appears to be severely limited, and thus its efficiency to maintain local populations of D. rapae in the vicinity of crops is questionable.


Association between morbidity and skeletal biomarkers of biological aging.
The Osseographic Scoring System (OSS) is a synthetic measure of skeletal aging that combines osteoporotic and osteoarthritic changes of the hand. The OSS is used to evaluate biological age in a population. The aim of the present study was to evaluate the association between certain chronic morbidities and the skeletal biomarker of biological aging. We performed a population-based study on a large sample of individuals who were receiving no medications to treat or prevent chronic morbidities [668 male (age 18-89 years) and 608 female (age 18-81 years) Chuvashians]. Morbidity data obtained from the individuals' medical records was sorted into 16 categories. A one-way ANOVA was used to elucidate the association between morbidity and age-adjusted OSS score. Statistically significant differences between means of OSS scores in affected vs. nonaffected individuals were found in the rheumatic diseases group. Ischemic heart disease, pulmonary diseases, traumatic brain injuries, and gynecological diseases also showed differences; however, after correction for multiple testing, the results were statistically nonsignificant. We conclude that individuals who suffer from the mentioned chronic morbidities will show a higher degree of skeletal aging. Further research is needed to clarify the biological mechanisms of association between certain types of morbidities and changes in skeletal aging.


Microfluidic Cell Transport with Piezoelectric Micro Diaphragm Pumps.
The automated transport of cells can enable far-reaching cell culture research. However, to date, such automated transport has been achieved with large pump systems that often come with long fluidic connections and a large power consumption. Improvement is possible with space- and energy-efficient piezoelectric micro diaphragm pumps, though a precondition for a successful use is to enable transport with little to no mechanical stress on the cell suspension. This study evaluates the impact of the microfluidic transport of cells with the piezoelectric micro diaphragm pump developed by our group. It includes the investigation of different actuation signals. Therewith, we aim to achieve optimal fluidic performance while maximizing the cell viability. The investigation of fluidic properties proves a similar performance with a hybrid actuation signal that is a rectangular waveform with sinusoidal flanks, compared to the fluidically optimal rectangular actuation. The comparison of the cell transport with three actuation signals, sinusoidal, rectangular, and hybrid actuation shows that the hybrid actuation causes less damage than the rectangular actuation. With a 5% reduction of the cell viability it causes similar strain to the transport with sinusoidal actuation. Piezoelectric micro diaphragm pumps with the fluidically efficient hybrid signal actuation are therefore an interesting option for integrable microfluidic workflows.


Ultra-high optical absorption efficiency from the ultraviolet to the infrared using multi-walled carbon nanotube ensembles.
The optical absorption efficiencies of vertically aligned multi-walled (MW)-carbon nanotube (CNT) ensembles are characterized in the 350-7000 nm wavelength range where CNT site densities > 1 × 10(11) /cm(2) are achieved directly on metallic substrates. The site density directly impacts the optical absorption characteristics, and while high-density arrays of CNTs on electrically insulating and non-metallic substrates have been commonly reported, achieving high site-densities on metals has been challenging and remains an area of active research. These absorber ensembles are ultra-thin (<10 μm) and yet they still exhibit a reflectance as low as ∼0.02%, which is 100 times lower than the reference; these characteristics make them potentially attractive for high-sensitivity and high-speed thermal detectors. In addition, the use of a plasma-enhanced chemical vapor deposition process for the synthesis of the absorbers increases the portfolio of materials that can be integrated with such absorbers due to the potential for reduced synthesis temperatures. The remarkable ruggedness of the absorbers is also demonstrated as they are exposed to high temperatures in an oxidizing ambient environment, making them well-suited for extreme thermal environments encountered in the field, potentially for solar cell applications. Finally, a phenomenological model enables the determinatiom of the extinction coefficients in these nanostructures and the results compare well with experiment.


Biological removal of carbon, nitrogen, and phosphorus in a sequencing batch reactor.
In this research the process performance of enhanced biological phosphorus removal was investigated in a sequencing batch reactor (SBR) having a new operational mode. The SBR system used in this study had simultaneous feeding and decanting conditions. The laboratory scale reactor (10 L) was operated for 392 days. The system was operated under 4 different sets each having 2 cycles per day. In each cycle, fill (4 h), anoxic (0.5 h), aerobic (7 h) and settling phases were present. In the fill phase, wastewater was fed from the bottom and the anoxic/anaerobic conditions were established in the settled sludge. During filling, the water left the system by water displacement. The system provided nitrification, denitrification as well as phosphorus and organic removal. High COD (90-98%), PO4-P (77-100%), and NH4-N (90-95%) removals were achieved by this system.


Lack of food effect on single-dose pharmacokinetics of brivanib, and safety and efficacy following multiple doses in subjects with advanced or metastatic solid tumors.
Brivanib alaninate, an orally available prodrug of brivanib, is currently under evaluation for the treatment of several malignancies. This study aimed to (1) investigate effects of a high-fat meal on single-dose pharmacokinetics of brivanib in subjects with advanced/metastatic solid tumors and (2) assess the safety and preliminary efficacy of single and multiple doses of brivanib alaninate in this population. A two-part study was conducted consisting of a single-dose phase (Part A) and a multiple-dose phase (Part B). In Part A, subjects received a single dose of brivanib alaninate (800 mg) either in a fasting state or following ingestion of a high-fat meal (approximately 951 kcal [15% protein, 33% carbohydrate, 52% fat]); serial blood samples were collected for pharmacokinetic analysis up to 48 h post-dosing. In Part B, subjects received brivanib alaninate (800 mg) once daily until discontinuation. Throughout both phases, subjects were evaluated for adverse events (AEs) and best clinical response. No clinically significant differences in brivanib exposure were observed between fed and fasting subjects in Part A; C (max) was unchanged and AUC(INF) decreased marginally when administered in a fed versus fasted state. In Part A, the incidence of treatment-emergent AEs was broadly similar in a fed or fasted state. Brivanib alaninate was generally well tolerated throughout the study and showed preliminary evidence of antitumor activity. Consumption of a high-fat meal had no significant effect on brivanib pharmacokinetics. The study further demonstrates the acceptable safety/tolerability profile and antitumor potential of brivanib in patients with advanced malignancies.


Efficacy of montelukast in the treatment of chronic obstructive pulmonary disease in five horses.
Five horses with moderate to severe chronic obstructive pulmonary disease (COPD) were treated with 0.11 (0.01) mg/kg bodyweight of montelukast, a cysteinyl leukotriene receptor antagonist, once a day for 26 days. The horses were evaluated clinically and endoscopically and subjected to arterial blood gas analysis and lung function tests before and after the period of treatment, and the plasma concentrations of montelukast were determined by high-performance liquid chromatography with fluorescence detection. The treatment did not result in statistically significant differences in the total scores of clinical and endoscopical signs, or in the difference in the arterioalveolar partial pressure of oxygen, maximal changes in pleural pressure, pulmonary resistance or dynamic compliance. The mean (sd) peak plasma concentration (C(max0) of montelukast was 12 (4) ng/ml and was reached 66 (13) minutes (t(max)) after its oral administration. The dose of montelukast per kg bodyweight was approximately the same as that for human beings, but the C(max) in the horses was 28 times lower and the t(max) was reached in one-fifth of the time, suggesting that its oral bioavailability may be lower.


Monostatic coaxial 1.5 μm laser Doppler velocimeter using a scanning Fabry-Perot interferometer.
We present a laser Doppler velocimeter (LDV) in monostatic coaxial arrangement consisting of off-the-shelf telecom-grade components: a single frequency laser (wavelength λ = 1.5 μm) and a high-finesse scanning Fabry-Perot interferometer (sFPI). In contrast to previous 1.5 μm LDV systems based on heterodyne detection, our sFPI-LDV has the advantages of having large remote sensing range not limited by laser coherence, high velocity dynamic range not limited by detector bandwidth and inherent sign discrimination of Doppler shift. The more optically efficient coaxial arrangement where transmitter and receiver optics share a common axis reduces the number of components and greatly simplifies the optical alignment. However, the sensitivity to unwanted backreflections is increased. To circumvent this problem, we employ a custom optical circulator design which compared to commercial fiber-optic circulator achieves ~40 dB reduction in strength of unwanted reflections (i.e. leakage) while maintaining high optical efficiency. Experiments with a solid target demonstrate the performance of the sFPI-LDV system with high sensitivity down to pW level at present update rates up to 10 Hz.


Problems with sexual function in people attending London general practitioners: cross sectional study.
To assess sexual behaviour, prevalence of ICD-10 diagnosed sexual dysfunction, associations between sexual and psychological problems, and help seeking for sexual problems in people attending general practice; to assess predictors of ICD-10 diagnosis of sexual dysfunction. Cross sectional study. 13 general practices in London. 1065 women and 447 men attending general practices. Prevalence and predictors of ICD-10 diagnoses of sexual dysfunction. 97 (22%, 95% confidence interval 18% to 25%) men and 422 (40%, 37% to 43%) women received at least one ICD-10 diagnosis, but only 3-4% had an entry relating to sexual problems in their general practice notes. The most common problems were erectile failure and lack or loss of sexual desire in men and lack or loss of sexual desire and failure of orgasmic response in women. Increasing age and being unemployed predicted sexual problems in women, and bisexual orientation, being non-white, and being unemployed were demographic predictors in men. No practice note factors predicted sexual problems in women, but high consulting rate predicted problems in men. The main clinical predictors were poor physical function and dissatisfaction with current sex life in both sexes and higher psychological morbidity in women. When all factors were considered, increasing age (odds ratio 1.01, 1.00 to 1.02), physical subscale score on the SF-12 (0.98, 0.97 to 0.99), sexual dissatisfaction (1.9, 1.5 to 2.4), and scoring over a 3/4 threshold score on the general health questionnaire (1.5, 1.1 to 1.9) independently predicted an ICD-10 sexual dysfunction diagnosis in women. Being bisexual (4.1, 1.3 to 12.8) was the only independent predictor of an ICD-10 diagnosis in men. Sexual difficulties are common in people attending general practitioners, and many people are prepared to talk about them with their doctors.


Low grade astrocytoma in children treated by surgery and radiation therapy.
Results of treatment of low grade astrocytoma by surgery and postoperative irradiation in 35 children under 15 were retrospectively analysed. The actuarial overall 5 and 10 year survival was 94 and 82 per cent, the actuarial relapse-free survival 87 and 70 per cent, respectively. The male patients fared better, although not significantly, than the females. No significant difference in relapse rate was found between cystic and solid tumors. The cystic processes had a more indolent course, but a fatal outcome was noted in one case (after 21 years). No relapse occurred in totally removed tumors (10 cases), which throws doubt on the need for postoperative irradiation in this subgroup. In the subtotally resected processes (24 cases) radiation therapy seemed to be beneficial, as in 3 of 4 local recurrences geographic misses of tumour remains probably existed. No dose response relationship was apparent. The performance status was severely impaired in 11 per cent (3/27 patients). Growth retardation was observed in 2 patients.


Ovarian stimulation and assisted reproductive technology outcomes in women transplanted with cryopreserved ovarian tissue: a systematic review.
To systematically review reproductive outcomes of assisted reproductive technology (ART) treatment in women transplanted with frozen-thawed ovarian tissue. Systematic review in accordance with guidelines from Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). Not applicable. Women undergoing ovarian tissue transplantation (OTT) and subsequent ART. Literature search in PubMed and Scopus databases. Time from OTT to initiation of ART, stimulation protocol, and conventional ART outcome measures. Twenty studies (including 15 case reports), specifying ART treatments and outcomes of 40 women undergoing OTT were identified. Multiple stimulation protocols were applied, with the modified natural cycle as the most frequently used. In total, 195 ART cycles were performed (4.0 cycles per patient) resulting in 1.5 follicles and 1.0 mature oocyte retrieved per cycle. Empty follicle rates ranged from 23% to 35% in the three largest cohort studies. Twenty-five women (62.5%) had one or more pregnancies, of which 28.6% were lost, resulting in a total of 20 live births (22 children). Overall the pregnancy rates varied from 3.9% to 19.3% and live-birth rates from 3.9% to 14.0% per cycle in the three cohort studies. Fertility treatment was initiated shortly after OTT in some centers, while others awaited natural conception before embarking on ART treatment. The reported pregnancy and live-birth rates for women undergoing OTT and ART were considerably lower than those of the general in vitro fertilization (IVF) population, corresponding to patients with poor ovarian reserve. In general, ART outcomes are underreported, and there is a lack of consensus regarding the timing of ART in relation to OTT and the type of ovarian stimulation protocol.


Hexagonal graphene onion rings.
Precise spatial control of materials is the key capability of engineering their optical, electronic, and mechanical properties. However, growth of graphene on Cu was revealed to be seed-induced two-dimensional (2D) growth, limiting the synthesis of complex graphene spatial structures. In this research, we report the growth of onion ring like three-dimensional (3D) graphene structures, which are comprised of concentric one-dimensional hexagonal graphene ribbon rings grown under 2D single-crystal monolayer graphene domains. The ring formation arises from the hydrogenation-induced edge nucleation and 3D growth of a new graphene layer on the edge and under the previous one, as supported by first principles calculations. This work reveals a new graphene-nucleation mechanism and could also offer impetus for the design of new 3D spatial structures of graphene or other 2D layered materials. Additionally, in this research, two special features of this new 3D graphene structure were demonstrated, including nanoribbon fabrication and potential use in lithium storage upon scaling.


Comparison of Fluvoxamine alone, Fluvoxamine and cognitive psychotherapy and psychotherapy alone in the treatment of panic disorder in Kelantan--implications for management by family doctors.
This paper reports the result of a brief therapy attempt at treating panic in a busy outpatient psychiatric clinic. The patients were cases of panic referred from the various outpatient clinics within the hospital complex. The patients were divided into three groups at random using one of three modalities of treatment, i.e. cognitive behaviour therapy (CBT), CBT and Fluvoxamine (FVX), and FVX alone. The therapy was aimed for a maximum of nine sessions after which the patients were to be discharged. There were 14 patients in each group. The results show that all the groups were similar in the severity and scores pre treatment but after the different types of treatment there was a significant difference among them. The FVX alone group, showed significant improvement from the pretreatment levels but did not show as much improvement as the other groups and the mean score was only 9.07 after nine sessions. The best group was the CBT in combination with FVX. This indicates that the best way to treat panic is to combine drug treatment and psychological treatment. It is also shown from the study that the combination group requires less FVX than the FVX alone group. This finding has implications for the treatment of panic at the family physician clinic.


[Evaluation of antioxidant vitamins intake by students of Warsaw University of Life Science].
The aim of this study was the assessment of antioxidant vitamins intake among students of the Faculty of Human Nutrition and Consumer Sciences of Warsaw University of Life Science. The investigation was done among 145 students at the age 21-28 in the 2008 year. Data about food intake were collected using a 3-day dietary records method. On this basis intake of vitamin A, beta-carotene, vitamin E and C was calculated The results shows that the quantity of antioxidant vitamins was adequate for vitamin C (64 mg/d, 102% EAR), but for vitamin A and E was higher then recommended value (698 microg retinol equivalent/d, 135% EAR and 11,5 mg tocopherol equivalent/d, 138% of AI). About 12-15% students used supplements, providing additional 574 microg vitamin A, 116 mg vitamin C and 4 mg vitamin E. It covered together with food intakes 260%, 298% EAR value and 199% of AI value, in case of vitamin E. In smokers, for which recommendations of intake antioxidant vitamins are slightly different, it was found too low their intake.


Critical role of active repression by E2F and Rb proteins in endoreplication during Drosophila development.
E2F transcription factors can activate or actively repress transcription of their target genes. The role of active repression during normal development has not been analyzed in detail. dE2F1(su89) is a novel allele of dE2F1 that disrupts dE2F1's association with RBF [the Drosophila retinoblastoma protein (Rb) homolog] but retains its transcription activation function. Interestingly, the dE2F1(su89) mutant, which has E2F activation by dE2F1(su89) and active repression by dE2F2, is viable and fertile with no gross developmental defects. In contrast, complete removal of active repression in de2f2;dE2F1(su89) mutants results in severe developmental defects in tissues with extensive endocycles but not in tissues derived from mitotic cycles. We show that the endoreplication defect resulted from a failure to downregulate the level of cyclin E during the gap phase of the endocycling cells. Importantly, reducing the gene dosage of cyclin E partially suppressed all the phenotypes associated with the endoreplication defect. These observations point to an important role for E2F-Rb complexes in the downregulation of cyclin E during the gap phase of endocycling cells in Drosophila development.


The role of tyrosine kinases in the pathogenesis of idiopathic pulmonary fibrosis.
Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease with a median survival time from diagnosis of 2-3 years. Although the pathogenic pathways have not been fully elucidated, IPF is believed to be caused by persistent epithelial injury in genetically susceptible individuals. Tyrosine kinases are involved in a range of signalling pathways that are essential for cellular homeostasis. However, there is substantial evidence from in vitro studies and animal models that receptor tyrosine kinases, such as the platelet-derived growth factor receptor, vascular endothelial growth factor receptor and fibroblast growth factor receptor, and non-receptor tyrosine kinases, such as the Src family, play critical roles in the pathogenesis of pulmonary fibrosis. For example, the expression and release of tyrosine kinases are altered in patients with IPF, while specific tyrosine kinases stimulate the proliferation of lung fibroblasts in vitro. Agents that inhibit tyrosine kinases have shown anti-fibrotic and anti-inflammatory effects in animal models of pulmonary fibrosis. Recently, the tyrosine kinase inhibitor nintedanib has shown positive results in two phase III trials in patients with IPF. Here, we summarise the evidence for involvement of specific tyrosine kinases in the pathogenesis of IPF and the development of tyrosine kinase inhibitors as treatments for IPF.


Spleen preserving distal pancreatectomy in an isolated blunt pancreatic trauma.
Blunt isolated pancreatic trauma is uncommon, accounting for 1%-4% of high impact abdominal injuries. In addition, its diagnosis can be difficult; physical signs may be poor and laboratory findings nonspecific, resulting in delayed treatment. Preserving the spleen during distal pancreatectomy (DP) is controversial. One of the spleen's functions regards immunity; complications following splenectomy include leukocytosis, thrombocytosis, overwhelming post splenectomy sepsis and some degree of immunodeficiency. This is why many authors favor its preservation. We describe a case of a young man with an isolated pancreatic trauma due to a blunt abdominal trauma with a delayed presentation who was treated with spleen-preserving DP and we discuss the value of this procedure with reference to the literature.


Silencing the Adipocytokine NOV: A Novel Approach to Reversing Oxidative Stress-Induced Cardiometabolic Dysfunction.
NOV/CCN3 is an adipocytokine recently linked to obesity, insulin resistance, and cardiometabolic dysfunction. NOV is manufactured and secreted from adipose tissue, with blood levels highly correlated with BMI. NOV levels are increased in obesity and a myriad of inflammatory diseases. Elevated NOV levels cause oxidative stress by increasing free radicals, decreasing antioxidants, and decreasing heme oxygenase (HO-1) levels, resulting in decreased vascular function. Silencing NOV in NOV knockout mice improved insulin sensitivity. We wanted to study how suppressing NOV expression in an obese animal model affected pathways and processes related to obesity, inflammation, and cardiometabolic function. This is the first study to investigate the interaction of adipose tissue-specific NOV/CCN3 and cardiometabolic function. We constructed a lentivirus containing the adiponectin-promoter-driven shNOV to examine the effect of NOV inhibition (shNOV) in adipose tissue on the heart of mice fed a high-fat diet. Mice were randomly divided into three groups (five per group): (1) lean (normal diet), (2) high-fat diet (HFD)+ sham virus, and (3) HFD + shNOV lentivirus. Blood pressure, tissue inflammation, and oxygen consumption were measured. Metabolic and mitochondrial markers were studied in fat and heart tissues. Mice fed an HFD developed adipocyte hypertrophy, fibrosis, inflammation, and decreased mitochondrial respiration. Inhibiting NOV expression in the adipose tissue of obese mice by shNOV increased mitochondrial markers for biogenesis (PGC-1α, the nuclear co-activator of HO-1) and functional integrity (FIS1) and insulin signaling (AKT). The upregulation of metabolic and mitochondrial markers was also evident in the hearts of the shNOV mice with the activation of mitophagy. Using RNA arrays, we identified a subgroup of genes that highly correlated with increased adipocyte mitochondrial autophagy in shNOV-treated mice. A heat map analysis in obese mice confirmed that the suppression of NOV overrides the genetic susceptibility of adiposity and the associated detrimental metabolic changes and correlates with the restoration of anti-inflammatory, thermogenic, and mitochondrial genes. Our novel findings demonstrate that inhibiting NOV expression improves adipose tissue function in a positive way in cardiometabolic function by inducing mitophagy and improving mitochondrial function by the upregulation of PGC-1α, the insulin sensitivity signaling protein. Inhibiting NOV expression increases PGC-1, a key component of cardiac bioenergetics, as well as key signaling components of metabolic change, resulting in improved glucose tolerance, improved mitochondrial function, and decreased inflammation. These metabolic changes resulted in increased oxygen consumption, decreased adipocyte size, and improved cardiac metabolism and vascular function at the structural level. The crosstalk of the adipose tissue-specific deletion of NOV/CCN3 improved cardiovascular function, representing a novel therapeutic strategy for obesity-related cardiometabolic dysfunction.


Noncatalytic domains of RGS9-1.Gbeta 5L play a decisive role in establishing its substrate specificity.
The complex between the photoreceptor-specific regulator of G protein signaling (RGS) protein, RGS9-1, and type 5 G protein beta-subunit, Gbeta5L, regulates the duration of the cellular response to light by stimulating the GTPase activity of G protein, transducin. An important property of RGS9-1.Gbeta5L is that it interacts specifically with transducin bound to its effector, cGMP phosphodiesterase, rather than with transducin alone. The minimal structure within the RGS9-1.Gbeta5L complex capable of activating transducin GTPase is the catalytic domain of RGS9. This domain itself is also able to discriminate between free and effector-bound transducin but to a lesser degree than RGS9-1.Gbeta5L. The goal of this study was to determine whether other, noncatalytic domains of RGS9-1.Gbeta5L enhance the intrinsic specificity of the catalytic domain or whether they set the specificity of RGS9-1.Gbeta5L regardless of the specificity of its catalytic domain. We found that a double L353E/R360P amino acid substitution reversed the specificity of the recombinant catalytic domain but did not reverse the specificity of RGS9-1.Gbeta5L. However, the degree of discrimination between free and effector-bound transducin was reduced. Therefore, noncatalytic domains of RGS9-1.Gbeta5L play a decisive role in establishing its substrate specificity, yet the high degree of this specificity observed under physiological conditions requires an additional contribution from the catalytic domain.


Use of computed tomography and plantar pressure measurement for management of neuropathic ulcers in patients with diabetes.
Total contact casting is effective at healing neuropathic ulcers, but patients have a high rate (30%-57%) of ulcer recurrence when they resume walking without the cast. The purposes of this case report are to describe how data from plantar pressure measurement and spiral x-ray computed tomography (SXCT) were used to help manage a patient with recurrent plantar ulcers and to discuss potential future benefits of this technology. The patient was a 62-year-old man with type 1 diabetes mellitus (DM) of 34 years' duration, peripheral neuropathy, and a recurrent plantar ulcer. Although total contact casting or relieving weight bearing with crutches apparently allowed the ulcer to heal, the ulcer recurred 3 times in an 18-month period. Spiral x-ray computed tomography and simultaneous pressure measurement were conducted to better understand the mechanism of his ulceration. The patient had a severe bony deformity that coincided with the location of highest plantar pressures (886 kPa). The results of the SXCT and pressure measurement convinced the patient to wear his prescribed footwear always, even when getting up in the middle of the night. The ulcer healed in 6 weeks, and the patient resumed his work, which required standing and walking for 8 to 10 hours a day. Following intervention, the patient's recurrent ulcer healed and remained healed for several months. Future benefits of these methods may include the ability to define how structural changes of the foot relate to increased plantar pressures and to help design and fabricate optimal orthoses.


COVID-19, ethics of care and feminist crisis management.
The COVID-19 pandemic threatens both lives and livelihoods. To reduce the spread of the virus, governments have introduced crisis management interventions that include border closures, quarantines, strict social distancing, marshalling of essential workers and enforced homeworking. COVID-19 measures are necessary to save the lives of some of the most vulnerable people within society, and yet in parallel they create a range of negative everyday effects for already marginalized people. Likely unintended consequences of the management of the COVID-19 crisis include elevated risk for workers in low-paid, precarious and care-based employment, over-representation of minority ethnic groups in case numbers and fatalities, and gendered barriers to work. Drawing upon feminist ethics of care, I theorize a radical alternative to the normative assumptions of rationalist crisis management. Rationalist approaches to crisis management are typified by utilitarian logics, masculine and militaristic language, and the belief that crises follow linear processes of signal detection, preparation/prevention, containment, recovery and learning. By privileging the quantifiable - resources and measurable outcomes - such approaches tend to omit considerations of pre-existing structural disadvantage. This article contributes a new theorization of crisis management that is grounded in feminist ethics to provide a care-based concern for all crisis affected people.


Treatment with recombinant interferon (alpha-2b) early after bone marrow transplantation in patients at high risk for relapse [corrected].
Relapse continues to be a problem after bone marrow transplantation (BMT) for hematologic malignancies, particularly in recipients of autologous or T-cell-depleted allogeneic grafts and in patients with advanced disease. Interferon (IFN) has shown antiproliferative activity in several malignant hematologic diseases and potentially may be of benefit when administered early after BMT when the number of residual cells is minimal. We tested in a phase I study the maximum tolerated daily dose of recombinant IFN alpha-2b in patients who had received a transplant for a disease at high risk for relapse (acute myeloid leukemia or non-Hodgkin's lymphoma beyond first remission, advanced myelodysplastic syndrome, acute lymphoblastic leukemia at any stage, chronic myeloid leukemia in accelerated or blast phase. Recombinant IFN alpha-2b was started at a dose of 0.5 x 10(6) IU/m2 and escalated by 0.5 x 10(6) IU/m2 in groups of three or four patients. The intention was to administer IFN as soon as stable engraftment after BMT was achieved (defined as an absolute neutrophil count of greater than 2.0 x 10(9)/L and platelet count greater than 100 x 10(9)/L for 5 consecutive days) and continued for 2 months. A total of 14 patients were enrolled after autologous (n = 3) or allogeneic (n = 11) BMT. Dose-limiting toxicity was myelosuppression. Significant (grade 2 to 4) neutropenia and thrombocytopenia led to discontinuation or dose reduction in five of eight patients receiving 1.5 x 10(6) or 2 x 10(6) IU/m2 IFN. Mild to moderate (grade 1 or 2) anorexia, weight loss, and fatigue occurred in the majority of patients independent of the IFN dose. De novo acute GVHD responsive to steroid treatment developed in 3 of 11 allograft recipients. Natural killer (NK) cell function was low before IFN treatment and was not improved with the cytokine. Conversely, interleukin-2-activated NK cells showed normal function even before starting IFN and no change was seen during IFN treatment. Clonogenic hematopoietic progenitor studies showed depression of all progenitor lines (colony-forming unit [CFU]-granulocyte, erythroid, monocyte, megakaryocyte, CFU granulocyte-macrophage, burst-forming unit-erythroid) by IFN at all dose levels except at 0.5 x 10(6) IU/m2. Considering this result and the incidence and severity of marrow depression seen at doses greater than 1.0 x 10(6) IU/m2, we would consider this the maximum dose safely tolerated if IFN alpha-2b is administered in this setting for a prolonged course on a daily basis.


The prevalence of Campylobacter amongst a free-range broiler breeder flock was primarily affected by flock age.
Campylobacter successfully colonizes broiler chickens, but little is known about the longer term natural history of colonization, since most flocks are slaughtered at an immature age. In this study, the prevalence and genetic diversity of Campylobacter colonizing a single free-range broiler breeder flock was investigated over the course of a year. The age of the flock was the most important factor in determining both the prevalence and diversity of Campylobacter over time. There was no correlation with season, temperature, the amount of rain and sunshine, or the dynamics of colonization amongst geographically and temporally matched broiler flocks. The higher prevalence rates coincided with the age at which broiler chickens are typically slaughtered, but then in the absence of bio-security or other intervention methods, and despite changes in flock management, the prevalence fell to significantly lower levels for the remainder of the study. The genetic diversity of Campylobacter increased as the flock aged, implying that genotypes were accumulated within the flock and may persist for a long time. A better understanding of the ecology of Campylobacter within commercial chicken flocks will allow the design of more effective farm-based interventions.


Preclinical evaluation of safety and immunogenicity of a primary series intranasal COVID-19 vaccine candidate (BBV154) and humoral immunogenicity evaluation of a heterologous prime-boost strategy with COVAXIN (BBV152).
Most if not all vaccine candidates developed to combat COVID-19 due to SARS-CoV-2 infection are administered parenterally. As SARS-CoV-2 is transmitted through infectious respiratory fluids, vaccine-induced mucosal immunity could provide an important contribution to control this pandemic. ChAd-SARS-CoV-2-S (BBV154), a replication-defective chimpanzee adenovirus (ChAd)-vectored intranasal (IN) COVID-19 vaccine candidate, encodes a prefusion-stabilized version of the SARS-CoV-2 spike protein containing two proline substitutions in the S2 subunit. We performed preclinical evaluations of BBV154 in mice, rats, hamsters and rabbits. Repeated dose toxicity studies presented excellent safety profiles in terms of pathology and biochemical analysis. IN administration of BBV154 elicited robust mucosal and systemic humoral immune responses coupled with Th1 cell-mediated immune responses. BBV154 IN vaccination also elicited potent variant (omicron) cross neutralization antibodies. Assessment of anti-vector (ChAd36) neutralizing antibodies following repeated doses of BBV154 IN administration showed insignificant titers of ChAd36 neutralizing antibodies. However, the immune sera derived from the same animals displayed significantly higher levels of SARS-CoV-2 virus neutralization (p<0.003). We also evaluated the safety and immunogenicity of heterologous prime-boost vaccination with intramuscular (IM) COVAXIN-prime followed by BBV154 IN administration. COVAXIN priming followed by BBV154 IN-booster showed an acceptable reactogenicity profile comparable to the homologous COVAXIN/COVAXIN or BBV154/BBV154 vaccination model. Heterologous vaccination of COVAXIN-prime and BBV154 booster also elicited superior (p<0.005) and cross variant (omicron) protective immune responses (p<0.013) compared with the homologous COVAXIN/COVAXIN schedule. BBV154 has successfully completed both homologous and heterologous combination schedules of human phase 3 clinical trials and received the restricted emergency use approval (in those aged above 18 years) from the Drugs Controller General of India (DCGI).


'Switch' technique to restore pronation and radial deviation in 17 patients with brachial plexus birth palsy.
We report outcomes of 17 children with brachial plexus birth palsy that underwent extensor carpi ulnaris to brachioradialis and brachioradialis to abductor pollicis longus transfers to correct supination and ulnar deviation deformity. Mean age at the time of surgery was 8.7 years and mean follow-up time was 21 months (8-44). These patients had marked increases in active forearm rotation and Canadian Occupational Performance Measure scores. Activity performance score on the measure increased from 1.1 to 6.7 and satisfaction score increased from 0.8 to 8.7 at final follow-up. We conclude from our patient series that these tendon transfers are effective in correction of the supination and ulnar deviation deformities in brachial plexus birth palsy. Level of evidence: IV.


Price-based promotions of alcohol: legislative consistencies and inconsistencies across the Australian retail, entertainment and media sectors.
Excessive alcohol consumption is a major public health issue internationally, with alcohol consumption being recognised as a leading cause of preventable illness and major social burden. To help ameliorate the risks and harms associated with alcohol consumption, all levels of governments have explored various legislative and regulatory provisions to support responsible alcohol consumption, service and promotion. In this article, using Australia as a case study, the legislative environment around responsible alcohol promotion and consumption across the Australian retail, entertainment and media sectors will be explored, with a focus on pricing and volume-based discounts. Whilst the potential harm and effect of both the licensed and non-licensed sectors appears to be widely acknowledged as similar in both scope and size of effect, legislative protections overwhelmingly focus solely on reducing the risks associated with alcohol consumption in licensed premises. This article explores the legislative provisions around preventing excessive alcohol consumption through promotional and marketing activities, and notes that whilst the licensed premises sector is facing increasing legislative restrictions, the off-premises sector remains unregulated and in some cases has even had existing restrictions removed, despite forming an increasing part of the alcohol chain in Australia. There appear to be inconsistencies and regulatory gaps in relation to price-based and volume-based discount alcohol promotions. Regulatory loopholes allow the retail sector in particular to use discounted alcohol as a promotional tool, in a way that is inconsistent with the goals of public health alcohol legislation, and in a way which would be illegal in any other sector. There appears to be a compelling case for introducing new restrictions, or extending existing restrictions, on these forms of promotion across all sectors involved in alcohol promotion, and there is considerable evidence that there would be considerable public health benefit from doing so.


Lung diffusing capacity for nitric oxide and carbon monoxide in relation to morphological changes as assessed by computed tomography in patients with cystic fibrosis.
Due to large-scale destruction, changes in membrane diffusion (Dm) may occur in cystic fibrosis (CF), in correspondence to alterations observed by computed tomography (CT). Dm can be easily quantified via the diffusing capacity for nitric oxide (DLNO), as opposed to the conventional diffusing capacity for carbon monoxide (DLCO). We thus studied the relationship between DLNO as well as DLCO and a CF-specific CT score in patients with stable CF. Simultaneous single-breath determinations of DLNO and DLCO were performed in 21 CF patients (mean +/- SD age 35 +/- 9 y, FEV1 66 +/- 28%pred). Patients also underwent spirometry and bodyplethysmography. CT scans were evaluated via the Brody score and rank correlations (rS) with z-scores of functional measures were computed. CT scores correlated best with DLNO (rS = -0.83; p < 0.001). Scores were also related to the volume-specific NO transfer coefficient (KNO; rS = -0.63; p < 0.01) and to DLCO (rS = -0.79; p < 0.001) but not KCO. Z-scores for DLNO were significantly lower than for DLCO (p < 0.001). Correlations with spirometric (e.g., FEV1, IVC) or bodyplethysmographic (e.g., SRaw, RV/TLC) indices were weaker than for DLNO or DLCO but most of them were also significant (p < 0.05 each). In this cross sectional study in patients with CF, DLNO and DLCO reflected CT-morphological alterations of the lung better than other measures. Thus the combined diffusing capacity for NO and CO may play a future role for the non-invasive, functional assessment of structural alterations of the lung in CF.


Phenotypic and Genomic Comparison of the Two Most Common ExoU-Positive Pseudomonas aeruginosa Clones, PA14 and ST235.
Genotyping of 2,882 Pseudomonas aeruginosa isolates that had been collected during the last 40 years identified the ExoU-positive lineages PA14 (ST253) and ST235 as the second and third most frequent clones in the P. aeruginosa population. Both clones were approximately 2-fold more frequently detected in animate habitats than in soil or aquatic habitats. While ST253 clone isolates were causing mainly acute and chronic infections in humans, ST235 isolates had been preferentially collected from hospitalized patients with severe acute infections, particularly, keratitis, urinary tract infections, burn wounds, and ventilator-associated pneumonia. The two major exoU clones differed substantially in the composition and flexibility of the accessory genome and by more than 8,000 amino acid sequences. Pronounced sequence variation between orthologs was noted in genes encoding elements of secretion systems and secreted effector molecules, including the type III secretion system, indicating the modes of action of the different clones. When comparing representatives of the two clones in batch culture, the PA14 strain orchestrated the quorum sensing circuitry for the expression of pathogenic traits and stopped growing in batch culture when it entered the stationary phase, but the quorum sensing-deficient ST235 strain expressed high type III secretion activity and continued to grow and to divide. In summary, unrestricted growth, high constitutive type III secretion activity, and facilitated uptake of foreign DNA could be major features that have made ST235 a global high-risk clone associated with poor outcomes of acute nosocomial infections.IMPORTANCE The ubiquitous and metabolically versatile environmental bacterium Pseudomonas aeruginosa can cause infections in a wide variety of hosts, including insects, plants, animals, and humans. P. aeruginosa is one of the ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) pathogens that are the major cause of nosocomial infections in the United States and are a threat all over the world because of their capacity to become increasingly resistant to all available antibiotics. Most experimental work on P. aeruginosa has been performed with reference strains PAO1 and PA14, providing deep insight into key metabolic and regulatory pathways thought to be applicable to all P. aeruginosa strains. However, this comparative study on the two most common exoU-positive clones taught us that there are major lineages in the population such as the global high-risk clone ST235 that exhibit uncommon traits of lifestyle, genome mobility, and pathogenicity distinct from those in our knowledge gained from the studies with the reference strains.


Association of early ambulation with length of stay and costs in total knee arthroplasty: retrospective cohort study.
The objective of this study was to evaluate the association of early ambulation with length of stay, costs, and outcomes in inpatients undergoing total knee arthroplasty. This is a retrospective study of 1504 patients who underwent total knee arthroplasty between August 2009 and January 2011 in a tertiary teaching hospital. All patients commenced physiotherapy interventions on postoperative day 1. The patients were categorized into an early ambulation group (began ambulating on postoperative day 1; n = 803) or a late ambulation group (began ambulating on postoperative day 2; n = 701). Multivariable regression and propensity score analyses were used to reduce selection biases. Early ambulation was associated with a statistically significant reduction in the adjusted average length of stay (-0.44 day; P < 0.001) and adjusted average total hospitalization costs (Singapore, -$385; United States, -$315; P < 0.001). Both groups did not differ significantly in the 90-day readmission rate; however, early ambulation was associated with higher odds of achieving at least 90 degrees of knee flexion (adjusted odds ratio, 1.33; P < 0.01) and requiring a walking aid with a smaller base of support (adjusted proportional odds ratio, 1.36; P < 0.001). As little as a 1-day difference in the day of first ambulation was associated with a shorter length of stay, lower hospitalization costs, and improved knee function. The results of this study provide the first empirical support for the usefulness of early ambulation after total knee arthroplasty.


Predicting 60-Day Mortality in a Home-Care Service: Development of a New Inter-RAI 49-Frailty Index in Patients With Chronic Disease and Without a Cancer Diagnosis.
Frailty Index (FI) is used to define the level of frailty in various clinical settings. Fifteen- and 26-item FIs have been demonstrated to predict 1-year mortality and intensity of care in home care (HC) and palliative home care (PHC). The objective of this study was to develop a new FI to predict the 60-day risk of death or transition to a PHC service after the initiation of an HC service in patients with chronic disease and without a cancer diagnosis. Retrospective cohort study. Patients 18 years and older followed in an HC service of a "Frailty Department-Local Palliative Care Network" from January 1, 2017, to October 31, 2021. A 49-item FI (FI-49) was developed selecting variables within the standardized international Residential Assessment Instrument assessments (interRAI-HC) and compared to existing FIs with 15 and 26 variables. A total of 2099 patients were included in the study with a median age of 80.0 years (IQR: 72.0-86.0) and a predominantly female population (62.4%). Among these patients, 8% died or were transferred to PHC within the 60-day follow-up. The FI-49 demonstrated a higher ability to predict 60-day mortality (C index 0.8165, 95% CI 0.7848-0.8481) compared to the 26- and 15-item FI. An FI-49 cutoff of 0.33 was also selected to provide clinicians with a more practical approach (C-index of 0.7044, 95% CI 0.6796-0.7292). The FI-49 is a good predictor of short-term mortality or transition to palliative care among older patients referred to an HC service. The automatic calculation of this tool could facilitate more appropriate care planning and the correct allocation of healthcare resources, especially considering the rapid ageing of the population.


The Seven Stages of Man: The Role of Developmental Stage on Medication Adherence in Respiratory Diseases.
The circumstances and drivers of the decision to initiate, implement, or persist with a medication differ for individuals at each developmental stage. For school-age children with asthma, the social environment of their family's cultural beliefs and the influence of peer networks and school policies are strong determinants of medication adherence. The stage of adolescence can be a particularly challenging time because there is a reduction in parental supervision of asthma management as the young person strives to become more autonomous. To illustrate the importance of such factors, adherence interventions in children and young adults with asthma have used peer-based supports and social supports, particularly social media platforms. In older patients, it is internal rather than external factors and age-related decline that pose challenges to medication adherence. Seniors face the challenges of polypharmacy, reduced social support, increased isolation, and loss of cognitive function. Strategies to promote adherence must be tailored to the developmental stage and respective behavioral determinants of the target group. This review considers the different attitudes toward medication and the different adherence behaviors in young and elderly patients with chronic respiratory conditions, specifically asthma and chronic obstructive pulmonary disease. Opportunities to intervene to optimize adherence are suggested.


Steroidal free radicals as possible intermediates in the biosynthesis of C19-delta 16-steroids.
Treatment of deoxycorticosterone with reagents that abstract a hydrogen atom from the C-21-hydroxyl group leads to the formation of androsta-4,16-dien-3-one. In vitro incubations of [1,2-(3)H]deoxycorticosterone with homogenates of boar testis tissue produce the tritiated dienone. This reaction can be formulated as an abstraction of hydrogen from the C-21-hydroxyl group to form a C-21-alkoxy radical, followed by a 1,5-H.shift from the C-16 atom and beta-scission of the C-16-alkoxy radical leading to the loss of a C(2) fragment (C-20 and C-21) with the formation of the double bond between C-16 and C-17. Both the chemical model and the in vitro experiment support further the thesis [Lieberman, S., L. Bandy, V. Lippman, and K. D. Roberts, Biochem. Biophys. Res. Commun., 34, 367 (1969)] that biosynthetic reactions in which molecular oxygen reacts with steroids (for example, side-chain cleavage or hydroxylation) involve steroidal free radicals.


Granuloma formation and host defense in chronic Mycobacterium tuberculosis infection requires PYCARD/ASC but not NLRP3 or caspase-1.
The NLR gene family mediates host immunity to various acute pathogenic stimuli, but its role in chronic infection is not known. This paper addressed the role of NLRP3 (NALP3), its adaptor protein PYCARD (ASC), and caspase-1 during infection with Mycobacterium tuberculosis (Mtb). Mtb infection of macrophages in culture induced IL-1beta secretion, and this requires the inflammasome components PYCARD, caspase-1, and NLRP3. However, in vivo Mtb aerosol infection of Nlrp3(-/-), Casp-1(-/-), and WT mice showed no differences in pulmonary IL-1beta production, bacterial burden, or long-term survival. In contrast, a significant role was observed for Pycard in host protection during chronic Mtb infection, as shown by an abrupt decrease in survival of Pycard(-/-) mice. Decreased survival of Pycard(-/-) animals was associated with defective granuloma formation. These data demonstrate that PYCARD exerts a novel inflammasome-independent role during chronic Mtb infection by containing the bacteria in granulomas.


Why outcomes measurement must be the basis for the development of clinical guidelines.
Three approaches to clinical quality improvement, health care standards, clinical guidelines, and quality improvement research, are differentiated by the quality of the available medical evidence linking health care processes to health care outcomes. The key determinant of Permanente Medical Group physician acceptance and use of a clinical guideline is the belief that solid data links the guideline's advice to improved patient outcomes. Purchasers, government agencies, regulatory organizations, and health plans must understand the difference between health care standards and clinical guidelines. Strict adherence to clinical guidelines should never be the basis for accountability for physicians, medical groups, or organized systems of care.


Decision Support Capabilities of Telemedicine in Emergency Prehospital Care: Systematic Review.
Telemedicine offers a unique opportunity to improve coordination and administration for urgent patient care remotely. In an emergency setting, it has been used to support first responders by providing telephone or video consultation with specialists at hospitals and through the exchange of prehospital patient information. This technological solution is evolving rapidly, yet there is a concern that it is being implemented without a demonstrated clinical need and effectiveness as well as without a thorough economic evaluation. Our objective is to systematically review whether the clinical outcomes achieved, as reported in the literature, favor telemedicine decision support for medical interventions during prehospital care. This systematic review included peer-reviewed journal articles. Searches of 7 databases and relevant reviews were conducted. Eligibility criteria consisted of studies that covered telemedicine as data- and information-sharing and two-way teleconsultation platforms, with the objective of supporting medical decisions (eg, diagnosis, treatment, and receiving hospital decision) in a prehospital emergency setting. Simulation studies and studies that included pediatric populations were excluded. The procedures in this review followed the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) statement. The Risk Of Bias In Non-randomised Studies-of Interventions (ROBINS-I) tool was used for the assessment of risk of bias. The results were synthesized based on predefined aspects of medical decisions that are made in a prehospital setting, which include diagnostic decision support, receiving facility decisions, and medical directions for treatment. All data extractions were done by at least two reviewers independently. Out of 42 full-text reviews, 7 were found eligible. Diagnostic support and medical direction and decision for treatments were often reported. A key finding of this review was the high agreement between prehospital diagnoses via telemedicine and final in-hospital diagnoses, as supported by quantitative evidence. However, a majority of the articles described the clinical value of having access to remote experts without robust quantitative data. Most telemedicine solutions were evaluated within a feasibility or short-term preliminary study. In general, the results were positive for telemedicine use; however, biases, due to preintervention confounding factors and a lack of documentation on quality assurance and protocol for telemedicine activation, make it difficult to determine the direct effect on patient outcomes. The information-sharing capacity of telemedicine enables access to remote experts to support medical decision making on scene or in prolonged field care. The influence of human and technology factors on patient care is poorly understood and documented.


Reelin immunoreactivity in neuritic varicosities in the human hippocampal formation of non-demented subjects and Alzheimer's disease patients.
Reelin and its downstream signaling members are important modulators of actin and microtubule cytoskeleton dynamics, a fundamental prerequisite for proper neurodevelopment and adult neuronal functions. Reductions in Reelin levels have been suggested to contribute to Alzheimer's disease (AD) pathophysiology. We have previously reported an age-related reduction in Reelin levels and its accumulation in neuritic varicosities along the olfactory-limbic tracts, which correlated with cognitive impairments in aged mice. Here, we aimed to investigate whether a similar Reelin-associated neuropathology is observed in the aged human hippocampus and whether it correlated with dementia status. Our immunohistochemical stainings revealed the presence of N- and C-terminus-containing Reelin fragments in corpora amylacea (CAm), aging-associated spherical deposits. The density of these deposits was increased in the molecular layer of the subiculum of AD compared to non-demented individuals. Despite the limitation of a small sample size, our evaluation of several neuronal and glial markers indicates that the presence of Reelin in CAm might be related to aging-associated impairments in neuronal transport leading to accumulation of organelles and protein metabolites in neuritic varicosities, as previously suggested by the findings and discussions in rodents and primates. Our results indicate that aging- and disease-associated changes in Reelin levels and proteolytic processing might play a role in the formation of CAm by altering cytoskeletal dynamics. However, its presence may also be an indicator of a degenerative state of neuritic compartments.


Identification of a highly immunoreactive epitope of Brugia malayi TPx recognized by the endemic sera.
Filarial thiordoxin peroxidase is a major antioxidant that plays a crucial role in parasite survival. Although Brugia malayi TPx has been shown to be a potential vaccine candidate, it shares 63% homology with its mammalian counterpart, limiting its use as a vaccine or drug target. In silico analysis of TPx sequence revealed a linear B epitope in the host's nonhomologous region. The peptide sequence (TPx peptide(27-48)) was synthesized, and its reactivity with clinical sera from an endemic region was analyzed. The peptide showed significantly high reactivity (P < 0.05) against the sera of putatively immune individuals compared to the nonendemic control sera. It also showed high reactivity against the sera of patients with chronic pathology and patent infection. The high reactivity of the peptide with endemic immune sera equivalent to that of whole protein shows that it forms a dominant B epitope of TPx protein and thus could be utilized for incorporation into a multiepitope vaccine construct for filariasis.


Metabolic recovery of adipose tissue is associated with improvement in insulin resistance in a model of experimental diabetes.
Obesity and insulin resistance are highly correlated with metabolic disturbances. Both the excess and lack of adipose tissue can lead to severe insulin resistance and diabetes. Adipose tissue plays an active role in energy homeostasis, hormone secretion, and other proteins that affect insulin sensitivity, appetite, energy balance, and lipid metabolism. Rats with streptozotocin-induced diabetes during the neonatal period develop the classic diabetic picture of hyperglycemia, hypoinsulinemia, and insulin resistance in adulthood. Low body weight and reduced epididymal (EP) fat mass were also seen in this model. The aim of this study was to investigate the glucose homeostasis and metabolic repercussions on the adipose tissue following chronic treatment with antidiabetic drugs in these animals. In the 4th week post birth, diabetic animals started an 8-week treatment with pioglitazone, metformin, or insulin. Animals were then killed, EP fat pads were excised, and blood samples were collected for biological and biochemical assays. Pioglitazone and insulin treatments, but not metformin, reduced hyperglycemia, polydipsia, and polyphagia. Although all antidiabetic therapies improved insulin sensitivity, this was particularly noteworthy in the pioglitazone-treated rats. Furthermore, a recovery of adipose mass and insulin levels were observed in pioglitazone- and insulin-, but not metformin-treated animals. Treatments with insulin or pioglitazone were able to correct significantly, but not completely, the metabolic abnormalities, parallel to full recovery of adipose mass, indicating that not only the low insulin levels but also the lack of adipose tissue might play a significant role on the pathophysiology of this particular diabetes model.


PREDICTIVE VALUE OF OPTICAL COHERENCE TOMOGRAPHIC FEATURES IN THE BEVACIZUMAB AND RANIBIZUMAB IN PATIENTS WITH DIABETIC MACULAR EDEMA (BRDME) STUDY.
To establish the predictive value of specific optical coherence tomography retinal features on visual outcomes and retinal thickness during anti-vascular endothelial growth factor treatment in patients with diabetic macular edema. Post hoc analysis of compound data of a prospective, 6-month, multicenter, randomized controlled trial of 119 patients with diabetic macular edema receiving either intravitreal bevacizumab or ranibizumab were analyzed to assess the associations between baseline retinal morphologic parameters and change in best-corrected visual acuity and central subfield thickness. Based on the study protocol of the core study, best-corrected visual acuity and central subfield thickness were obtained before each mandatory monthly injection during 6 months. The presence of serous retinal detachment at baseline was associated with significant improvement in best-corrected visual acuity letter score at Month 3 and Month 6 (P < 0.001 and P = 0.01, respectively). In addition, the presence of disorganization of retinal inner layers was associated with lower best-corrected visual acuity letter score at Month 3 and Month 6 (P < 0.05 and P = 0.01, respectively). This study found that serous retinal detachment and disorganization of retinal inner layers were associated with different treatment responses to anti-vascular endothelial growth factor therapy in patients with diabetic macular edema.


Nontyphoidal Salmonella Osteomyelitis in Immunocompetent Children Without Hemoglobinopathies: A Case Series and Systematic Review of the Literature.
Salmonella osteomyelitis is well-described in children with hemoglobinopathies, particularly infection with Salmonella typhi. To characterize nontyphoidal osteomyelitis in otherwise healthy children without hemoglobinopathies, we performed a retrospective review of children discharged from our institution with this condition, supplemented with a systematic literature review. Among the 46 subjects identified, common risk factors for Salmonella infection were frequently absent and complications were common.


Building consensus for provision of breathlessness rehabilitation for patients with chronic obstructive pulmonary disease and chronic heart failure.
The study aimed to gain consensus on key priorities for developing breathlessness rehabilitation services for patients with chronic obstructive pulmonary disease (COPD) and chronic heart failure (CHF). Seventy-four invited stakeholders attended a 1-day conference to review the evidence base for exercise-based rehabilitation in COPD and CHF. In addition, 47 recorded their views on a series of statements regarding breathlessness rehabilitation tailored to the needs of both patient groups. A total of 75% of stakeholders supported symptom-based rather than disease-based rehabilitation for breathlessness with 89% believing that such services would be attractive for healthcare commissioners. A total of 87% thought patients with CHF could be exercised using COPD training principles and vice versa. A total of 81% felt community-based exercise training was safe for patients with severe CHF or COPD, but only 23% viewed manual-delivered rehabilitation an effective alternative to supervised exercise training. Although there was strong consensus that exercise training was a core component of rehabilitation in CHF and COPD populations, only 36% thought that this was the 'most important' component, highlighting the need for psychological and other non-exercise interventions for breathlessness. Patients with COPD and CHF face similar problems of breathlessness and disability on a background of multi-morbidity. Existing pulmonary and cardiac rehabilitation services should seek synergies to provide sufficient flexibility to accommodate all patients with COPD and CHF. Development of new services could consider adopting a patient-focused rather than disease-based approach. Exercise training is a core component, but rehabilitation should include other interventions to address dyspnoea, psychological and education needs of patients and needs of carers.


[The current views on the role and importance of phytotherapy in the structure of scientific medicine].
Phytotherapy, in spite of a long-continued employment in empirical and scientific medicine, does not have distinct scientific doctrine, conceptual strategies and tactics of use in practical treatment. Ways and means of phytotherapy are more often than not regarded as adjuncts to pharmacotherapy. In connection with formation during recent decades of new concepts of pathogenetic essence of pathological processes as of combined disturbances in biochemical reactions,--at the membrane and cell level in the first place, the role and value of phytotherapy as an independent method and means of metabolic therapy is considered in a new perspective.


Adaptation of WHO's generic tuberculosis patient cost instrument for a longitudinal study in Africa.
The WHO developed a generic 'TB patient cost survey' tool and a standardized approach to assess the direct and indirect costs of TB incurred by patients and their households, estimate the proportion of patients experiencing catastrophic costs, and measure the impact of interventions to reduce patient costs. While the generic tool is a facility-based cross-sectional survey, this standardized approach needs to be adapted for longitudinal studies. A longitudinal approach may overcome some of the limitations of a cross-sectional design and estimate the economic burden of TB more precisely. We describe the process of creating a longitudinal instrument and its application to the TB Sequel study, an ongoing multi-country, multi-center observational cohort study. We adapted the cross-sectional WHO generic TB patient cost survey instrument for the longitudinal study design of TB Sequel and the local context in each study country (South Africa, Mozambique, Tanzania, and The Gambia). The generic instrument was adapted for use at enrollment (start of TB treatment; Day 0) and at 2, 6, 12 and 24 months after enrollment, time points intended to capture costs incurred for diagnosis, during treatment, at the end of treatment, and during long-term follow-up once treatment has been completed. These time points make the adapted version suitable for use in patients with either drug-sensitive or drug-resistant TB. Using the adapted tool provides the opportunity to repeat measures and make comparisons over time, describe changes that extend beyond treatment completion, and link cost survey data to treatment outcomes and post-TB sequelae. Trial registration: ClinicalTrials.gov: NCT032516 August 1196, 2017. Abbreviations: DOTS: Directly observed treatment, short-course; DR-TB: Drug-resistant tuberculosis; MDR-TB: Multi-drug resistant tuberculosis; NTP: National Tuberculosis Programme; TB: Tuberculosis; USD: United States Dollar; WHO: World Health Organization.


Montelukast Does not Maintain Symptom Remission After Topical Steroid Therapy for Eosinophilic Esophagitis.
Montelukast, a cysteinyl leukotriene type-1 receptor blocker, has been shown in small retrospective studies to reduce symptoms in patients with eosinophilic esophagitis (EoE). We performed a randomized, placebo-controlled, double-blind trial to determine whether montelukast maintains symptomatic remission induced by topical steroid therapy in patients with EoE. We performed a prospective study of adult patients with EoE (solid-food dysphagia and a peak esophageal eosinophil count of >20 cells/high-powered field) enrolled at the Mayo Clinic in Rochester, Minnesota, from April 2008 through February 2015. All patients had been treated previously for at least 6 weeks with a topical steroid until their symptoms were in remission. Steroids were discontinued and patients then were assigned randomly to groups given montelukast (20 mg/day, n = 20) or placebo (n = 21) for 26 weeks (groups were matched for age, sex, history of allergic disease, reflux symptoms, and endoscopic findings of EoE). Study participants were assessed via a structured telephone interview at weeks 2, 4, 8, 12, 16, 20, and 24. Remission was defined as the absence of solid-food dysphagia. Based on an intention-to-treat analysis, after 26 weeks, 40.0% of subjects in the montelukast group and 23.8% in the placebo group were in remission. The odds ratio for remission in the montelukast group was 0.48 (95% confidence interval, 0.10-2.16) (P = .33). No side effects were reported from either group. In a randomized controlled trial of the ability of montelukast to maintain remission in patients in remission from EoE after steroid therapy, we found montelukast to be well tolerated; 40% of patients remained in remission, but this proportion did not differ significantly from that of the placebo group. ClinicalTrials.gov no: NCT00511316.


Glucose-6-phosphate dehydrogenase and MEG3 controls hypoxia-induced expression of serum response factor (SRF) and SRF-dependent genes in pulmonary smooth muscle cell.
Although hypoxia induces aberrant gene expression and dedifferentiation of smooth muscle cells (SMCs), mechanisms that alter dedifferentiation gene expression by hypoxia remain unclear. Therefore, we aimed to gain insight into the hypoxia-controlled gene expression in SMCs. We conducted studies using SMCs cultured in 3% oxygen (hypoxia) and the lungs of mice exposed to 10% oxygen (hypoxia). Our results suggest hypoxia upregulated expression of transcription factor CP2-like protein1, krüppel-like factor 4, and E2f transcription factor 1 enriched genes including basonuclin 2 (Bcn2), serum response factor (Srf), polycomb 3 (Cbx8), homeobox D9 (Hoxd9), lysine demethylase 1A (Kdm1a), etc. Additionally, we found that silencing glucose-6-phosphate dehydrogenase (G6PD) expression and inhibiting G6PD activity downregulated Srf transcript and hypomethylation of SMC genes (Myocd, Myh11, and Cnn1) and concomitantly increased their expression in the lungs of hypoxic mice. Furthermore, G6PD inhibition hypomethylated MEG3, a long non-coding RNA, gene and upregulated MEG3 expression in the lungs of hypoxic mice and in hypoxic SMCs. Silencing MEG3 expression in SMC mitigated the hypoxia-induced transcription of SRF. These findings collectively demonstrate that MEG3 and G6PD codependently regulate Srf expression in hypoxic SMCs. Moreover, G6PD inhibition upregulated SRF-MYOCD-driven gene expression, determinant of a differentiated SMC phenotype.


Project HeartBeat! Concept, development, and design.
Major cardiovascular disease (CVD) risk factors begin development in childhood and adolescence. Project HeartBeat! studied early development of these risk factors as growth processes. Growth, body composition, sexual maturation, major CVD risk factors, and cardiac structure and function were monitored every 4 months for up to 4 years among 678 children and adolescents (49.1% girls; 20.1% blacks) aged 8, 11, or 14 years at study entry. All resided in The Woodlands or Conroe TX. Interviews were conducted at entry and annually on diet, physical activity, and health history of participants and their families. Data were collected from 1991 to 1995, and study investigators continue data analysis and reporting. Overlap in ages at examination among three cohorts (aged 8-12, 11-15, and 14-18 years at baseline) and use of multilevel modeling methods permit analysis of some 5500 observations on each principal variable for the synthetic cohort from ages 8 to 18 years. The mixed-longitudinal design provides trajectories of change with age, for total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and triglycerides; systolic, and fourth-phase and fifth-phase diastolic blood pressure, and left ventricular mass. These trajectories are then related to concurrent measures of multiple indices of body composition and sexual maturation and adjusted for energy intake and physical activity. The data provide valuable insights into risk factor development and suggest a fresh approach to understanding influences on blood lipids, blood pressure, and left ventricular mass during the period of childhood and adolescence, a period of dynamic change in these risk factors.


Factor XI, but not prekallikrein, blocks high molecular weight kininogen binding to human umbilical vein endothelial cells.
Previous studies on the interaction of high molecular weight kininogen (HK) with endothelial cells have reported a large number of binding sites (106-107 sites/cell) with differing relative affinities (KD = 7-130 nm) and have implicated various receptors or receptor complexes. In this study, we examined the binding of HK to human umbilical vein endothelial cells (HUVEC) with a novel assay system utilizing HUVEC immobilized on microcarrier beads, which eliminates the detection of the high affinity binding sites found nonspecifically in conventional microtiter well assays. We report that HK binds to 8.5 x 104 high affinity (KD = 21 nm) sites per HUVEC, i.e. 10-100-fold fewer than previously reported. Although HK binding is unaffected by the presence of a physiological concentration of prekallikrein, factor XI abrogates HK binding to HUVEC in a concentration-dependent manner. Disruption of the naturally occurring complex between factor XI and HK by the addition of a 31-amino acid peptide mimicking the factor XI-binding site on HK restored HK binding to HUVEC. Furthermore, HK inhibited thrombin-stimulated von Willebrand factor release by HUVEC but not thrombin receptor activation peptide (SFLLRN-amide)-stimulated von Willebrand factor release. Factor XI restored the ability of thrombin to stimulate von Willebrand factor release in the presence of low HK concentrations. These results suggest that free HK, or HK in complex with prekallikrein but not in complex with factor XI, interacts with the endothelium and can maintain endothelial cell quiescence by preventing endothelial stimulation by thrombin.


Time-domain discrete-dipole approximation for simulation of temporal response of plasmonic nanoparticles.
We develop the time-domain discrete dipole approximation (DDA), describing the temporal evolution of electric field in plasmonic nanostructures. The main equation is obtained by taking the inverse Fourier transform of the Taylor expansion of the frequency-domain DDA in terms of frequency deviation from the central frequency. Thus we assume that incident wavefronts of different frequencies accumulate relatively small phase difference when passing the particle. This assumption is always valid for nanoparticles much smaller than the wavelength. Being the time-domain method, the proposed approach also requires an analytic frequency dependence of electric permittivity, e.g. the Drude model. We present numerical results of application of the time-domain DDA to silver nanosphere, rod, and disk, which agree well with that obtained with its frequency-domain counterpart and the finite-difference time-domain method. Moreover, the time-domain DDA is the fastest of the three methods for incident pulses of several-femtoseconds width. Thus, it can effectively be applied for modeling the temporal responses of plasmonic nanostructures.


Modelling of healthcare-associated infections: a study on the dynamics of pathogen transmission by using an individual-based approach.
Prevention and control of Healthcare Associated Infections (HAIs) has become a high priority for most healthcare organizations. Mathematical models can provide insights into the dynamics of nosocomial infections and help to evaluate the effect of infection control measures. The model presented in this paper adopts an individual-based and stochastic approach to investigate MRSA outbreaks in a hospital ward. A computer simulation was implemented to analyze the dynamics of the system associated with the spread of the infection and to carry out studies on space and personnel management. This study suggests that a strict spatial cohorting might be ineffective, if it is not combined with personnel cohorting.


AAV capsid bioengineering in primary human retina models.
Adeno-associated viral (AAV) vector-mediated retinal gene therapy is an active field of both pre-clinical as well as clinical research. As with other gene therapy clinical targets, novel bioengineered AAV variants developed by directed evolution or rational design to possess unique desirable properties, are entering retinal gene therapy translational programs. However, it is becoming increasingly evident that predictive preclinical models are required to develop and functionally validate these novel AAVs prior to clinical studies. To investigate if, and to what extent, primary retinal explant culture could be used for AAV capsid development, this study performed a large high-throughput screen of 51 existing AAV capsids in primary human retina explants and other models of the human retina. Furthermore, we applied transgene expression-based directed evolution to develop novel capsids for more efficient transduction of primary human retina cells and compared the top variants to the strongest existing benchmarks identified in the screening described above. A direct side-by-side comparison of the newly developed capsids in four different in vitro and ex vivo model systems of the human retina allowed us to identify novel AAV variants capable of high transgene expression in primary human retina cells.


The marginating-pulmonary immune compartment in rats: characteristics of continuous inflammation and activated NK cells.
A significant role has been indicated for cellular immunity in controlling circulating cancer cells, but most autologous tumor cells seem resistant, in vitro, to natural killer cell (NKC) and cytotoxic T lymphocytes cytotoxicity. Addressing this apparent contradiction, we recently identified a unique leukocyte population, marginating-pulmonary (MP)-leukocytes, which exhibit potent natural killer (NK) cytotoxicity. Here, we characterize the MP-compartment in naive and immunostimulated rats, and assessed its cytotoxicity against "NK-resistant" tumors cells. Animals were treated with poly I-C (3x0.2 mg/kg) or saline, and circulating-leukocytes and MP-leukocytes were collected and analyzed in terms of cellular composition, cellular activation markers, and NK cytotoxicity of leukocytes and purified NKCs. Compared with circulating-leukocytes, MP-leukocytes showed greater proportion of granulocytes, monocytes, NKCs, and large NKCs; higher expression of activation and adhesion markers (CD25, CD11a, CD11b, and NKR-P1, IFN-gamma); and elevated NK cytotoxicity of leukocytes and purified NKCs against several syngeneic and xenogeneic NK-resistant target cells (from both F344 and BDX inbred rats). In immunostimulated animals (treated with poly I-C), but not in naive animals, purified NKCs from the MP-compartment showed markedly superior cytotoxicity, suggesting that poly I-C immunostimulation uniquely affect MP-NKCs, and that in naive animals other MP-leukocytes support NK cytotoxicity. Overall, the results suggest that the MP-compartment is characterized by a continuous activated inflammatory microenvironment uniquely affected by immunostimulation. If similarly potent MP-NKCs exist in patients, then circulating autologous tumor cells that are considered "NK-resistant" could actually be controlled by MP-NKCs. Innate immunity may assume greater role in controlling malignant spread, especially after immunostimulation.


Hepatic function and non-invasive hepatosplanchnic monitoring in patients with abdominal hypertension.
A better understanding of intra-abdominal hypertension with relation to the liver is vital to the management of all forms of liver pathophysiology. Supporting good hepatic function within the critically ill patient is important not only in maintaining synthetic function, but also in avoiding the multi-organ complications of liver dysfunction. The resulting reduction in hepato-splanchnic blood flow (HSBF) observed with increasing intra-abdominal pressure has been clearly documented and seen to be exaggerated in animals with established liver disease. Unfortunately the tools required to measure this, remain difficult to apply routinely in the clinical setting and as such goal directed therapy to specifically improve the hepatosplanchnic circulation remains elusive. Given the documented effects of IAP on HSBF and the relatively high incidence of intra-abdominal hypertension and the abdominal compartment syndrome within "liver patients" as a whole, close attention to IAP and timely correction by appropriate medical or surgical means would appear to be essential.


A blast wave from the 1843 eruption of eta Carinae.
Very massive stars shed much of their mass in violent precursor eruptions as luminous blue variables (LBVs) before reaching their most likely end as supernovae, but the cause of LBV eruptions is unknown. The nineteenth-century eruption of eta Carinae, the prototype of these events, ejected about 12 solar masses at speeds of 650 km s(-1), with a kinetic energy of almost 10(50) erg (ref. 4). Some faster material with speeds up to 1,000-2,000 km s(-1) had previously been reported but its full distribution was unknown. Here I report observations of much faster material with speeds up to 3,500-6,000 km s(-1), reaching farther from the star than the fastest material in previous reports. This fast material roughly doubles the kinetic energy of the nineteenth-century event and suggests that it released a blast wave now propagating ahead of the massive ejecta. As a result, eta Carinae's outer shell now mimics a low-energy supernova remnant. The eruption has usually been discussed in terms of an extreme wind driven by the star's luminosity, but the fast material reported here indicates that it may have been powered by a deep-seated explosion rivalling a supernova, perhaps triggered by the pulsational pair instability. This may alter interpretations of similar events seen in other galaxies.


Nano-Iron Oxide (Fe3O4) Mitigates the Effects of Microplastics on a Ryegrass Soil-Microbe-Plant System.
To understand microplastic-nanomaterial interactions in agricultural systems, a randomized block 90-day pot experiment was set up to cultivate ryegrass seedings in a typical red sandy soil amended with compost (1:9 ratio). Polyvinyl chloride (PVC) and polyethylene (PE) microplastic (MP) contaminants were added into pot soils at 0.1 and 10%, whereas nano-Fe3O4 (as nanoenabled agrochemicals) was added at 0.1% and 0.5% in comparison with chemical-free controls. The combination of nano-Fe3O4 and MPs significantly increased the soil pH (+3% to + 17%) but decreased the total nitrogen content (-9% to - 30%; P < 0.05). The treatment group with both nano-Fe3O4 and PE had the highest total soil C (29 g kg-1 vs 20 g kg-1 in control) and C/N ratio (13 vs 8 in control). Increased rhizosphere nano-Fe3O4 concentrations promoted ryegrass growth (+42% dry weight) by enhancing the chlorophyll (+20%) and carotenoid (+15%) activities. Plant leaf and root peroxidase enzyme activity was more significantly affected by nano-Fe3O4 with PVC (+15%) than with PE (+6%). Nano-Fe3O4 significantly changed the ryegrass bacterial community structure from belowground (the rhizoplane and root endosphere) to aboveground (the phylloplane). Under MP contamination, the addition of nano-Fe3O4 increased bacterial diversity (+0.35%) and abundance (+30%) in the phylloplane and further intensified the connectivity of ryegrass aboveground bacterial networks (positive association increased 17%). The structural equation model showed that the change in the plant microbiome was associated with the rhizosphere microbiome. Overall, these findings imply the positive influences of nano-Fe3O4 on the soil-microbe-plant system and establish a method to alleviate the harmful effects of MP accumulation in soils.


Oriented cell motility and division underlie early limb bud morphogenesis.
The vertebrate limb bud arises from lateral plate mesoderm and its overlying ectoderm. Despite progress regarding the genetic requirements for limb development, morphogenetic mechanisms that generate early outgrowth remain relatively undefined. We show by live imaging and lineage tracing in different vertebrate models that the lateral plate contributes mesoderm to the early limb bud through directional cell movement. The direction of cell motion, longitudinal cell axes and bias in cell division planes lie largely parallel to one another along the rostrocaudal (head-tail) axis in lateral plate mesoderm. Transition of these parameters from a rostrocaudal to a mediolateral (outward from the body wall) orientation accompanies early limb bud outgrowth. Furthermore, we provide evidence that Wnt5a acts as a chemoattractant in the emerging limb bud where it contributes to the establishment of cell polarity that is likely to underlie the oriented cell behaviours.


A YSK-Type Dehydrin from Nicotiana tabacum Enhanced Copper Tolerance in Escherichia coli.
Copper is an essential micronutrient for the maintenance of normal cell function but is toxic in excess. Dehydrins are group two late embryogenesis abundant proteins, which facilitate plant survival in harsh environmental conditions. Here, a YSK-type dehydrin, NtDhn17, was cloned from Nicotiana tabacum under copper toxicity and characterized using a heterologous expression system and in vitro or in vivo experiments and exhibited characteristics of intrinsic disorder during in vitro analyses. Heterologous expression of NtDHN17 enhanced the tolerance of E. coli to various metals, osmotic, and oxidative stress. NtDHN17 showed no Cu2+-binding properties in vivo or in vitro, indicating that metal ion binding is not universal among dehydrins. In vitro and in vivo experiments suggested that NtDHN17 behaved as a potent anti-aggregation agent providing strong protection to aggregated proteins induced by excess copper ions, an effect dependent on the K-segment but not on the Y- or S-segments. In summary, the protective role of NtDHN17 towards E. coli under conditions of copper toxicity may be related to anti-aggregation ability rather than its acting as an ion scavenger, which might be a valuable target for the genetic improvement of resistance to heavy metal stresses in plants.