Time-of-Flight MR Angiography for Detection of Cerebral Hyperperfusion Syndrome after Superficial Temporal Artery-Middle Cerebral Artery Anastomosis in Moyamoya Disease.
Cerebral hyperperfusion syndrome is a potential complication of superficial temporal artery-MCA anastomosis for Moyamoya disease. In this study, we evaluated whether TOF-MRA could assess cerebral hyperperfusion syndrome after superficial temporal artery-MCA anastomosis for this disease. This retrospective study included patients with Moyamoya disease who underwent superficial temporal artery-MCA single anastomosis. TOF-MRA and SPECT were performed before and 1-6 days after anastomosis. Bilateral ROIs on the source image of TOF-MRA were manually placed directly on the parietal branch of the superficial temporal artery just after branching the frontal branch of the superficial temporal artery and on the contralateral superficial temporal artery on the same axial image, respectively. The change ratio of the maximum signal intensity of the superficial temporal artery on TOF-MRA was calculated by using the following formula: (Postoperative Ipsilateral/Postoperative Contralateral)/(Preoperative Ipsilateral/Preoperative Contralateral). Of 23 patients (26 sides) who underwent the operation, 5 sides showed cerebral hyperperfusion syndrome postoperatively. There was a significant difference in the change ratio of signal intensity on TOF-MRA observed between the cerebral hyperperfusion syndrome and non-cerebral hyperperfusion syndrome groups (cerebral hyperperfusion syndrome group: 1.88 ± 0.32; non-cerebral hyperperfusion syndrome group: 1.03 ± 0.20; P = .0009). The minimum ratio value for the cerebral hyperperfusion syndrome group was 1.63, and the maximum ratio value for the non-cerebral hyperperfusion syndrome group was 1.30. Thus, no overlap was observed between the 2 groups for the change ratio of signal intensity on TOF-MRA. Diagnosis of cerebral hyperperfusion syndrome is indicated by an increase in the change ratio of signal intensity on TOF-MRA by more than approximately 1.5 times the preoperative levels.


Coronary revascularization for patients with severe left ventricular dysfunction.
The efficacy of coronary artery bypass graft surgery (CABG) and percutaneous coronary intervention (PCI) in patients with coronary artery disease has been well defined by randomized controlled trials. However, patients with severe left ventricular dysfunction (ejection fraction <35%) were underrepresented in these trials, and management of these complex patients remains unclear. The purpose of this study was to compare the outcomes of patients with coronary artery disease and left ventricular dysfunction undergoing CABG versus PCI. The Alberta Provincial Project for Outcome Assessment in Coronary Heart Disease (APPROACH), a clinical data collection and outcome monitoring initiative for the province of Alberta, Canada, was used to identify 2925 patients with coronary artery disease and left ventricular dysfunction undergoing CABG (n = 1,326) or PCI (n = 1,599) between 1995 and 2008. Patients were propensity matched to obtain comparable subgroups among left ventricular dysfunction patients. Cox proportional hazard analysis of the propensity-matched subgroups identified that CABG was significantly associated with lower rates of repeat revascularization and better survival compared with PCI at 1, 5, 10, and 15 years. Other significant independent predictors of poor long-term survival included age, renal failure, heart failure, diabetes mellitus, peripheral vascular disease, prior myocardial infarction, left main coronary artery disease, and prior CABG. For patients with coronary artery disease and left ventricular dysfunction, CABG was associated with lower rates of repeat revascularization and improved survival over PCI, after adjustment for baseline risk profile differences. Further research exploring the factors leading to use of a particular revascularization modality in this patient population is required.


The supplementation of L-carnitine in septic shock patients: Systematic review and meta-analysis.
Sepsis and septic shock are severe and difficult-to-treat conditions with high lethality. There is interest in identifying new adjunct therapies that are effective in reducing mortality. In this context, L-carnitine has been investigated in trials as a potentially beneficial drug. Therefore, the aim of this systematic review was to assess the clinical evidence to support the use of L-carnitine in septic shock patients to reduce the risk of mortality. The objective of this review was to evaluate the effect of L-carnitine compared to placebo or Usual Care (UC) on the mortality rate in hospitalized adult septic shock patients. The authors exclusively included randomized clinical trials that compared the use of L-carnitine versus placebo in adult (> 18 years old) septic shock patients. The outcome was a mortality rate of 28 days. This systematic review and meta-analysis were performed following the PRISMA guidelines and registered in PROSPERO with the ID CRD42020180499. Following the initial search, 4007 citations were identified, with 2701 remaining after duplicate removal. Eight citations were selected for body text reading, and two were selected for inclusion. The studies enrolled 275 patients, with 186 in the carnitine arm and 89 in the placebo arm. The effect of L-carnitine uses in septic shock patients showed a difference risk of -0.03 (95% Confidence Interval: -0.15-0.10, I2 = 77%, p = 0.69) compared to placebo/in mortality rate with low quality of evidence. There is low-quality evidence that the use of L-carnitine has no significant effect on reducing 28-day mortality in septic shock patients.


Psychometric properties of the Need for Recovery after work scale: test-retest reliability and sensitivity to detect change.
Monitoring worker health and evaluating occupational healthcare interventions requires sensitive instruments that are reliable over time. The Need for Recovery scale (NFR), which quantifies workers' difficulties in recovering from work related exertions, may be a relevant instrument in this respect. To examine (1) the NFR's test-retest reliability and (2) the NFR's sensitivity to detect the effect of a fatigue inducing change, namely an increase in working hours. Two year longitudinal data of 526 truck drivers and 144 nurses were used. Two week, one year, and two year test-retest reliability was examined in both stable and unstable work environments by calculating intraclass correlations (ICCs). Work environmental (in)stability was quantified by four events that might have occurred during the follow up period: (1) a reorganisation or merge (0 = yes, 1 = no), (2) a change of supervisor or management (0 = yes, 1 = no), (3) a change in working hours or work schedules (0 = yes, 1 = no), and (4) a change in work activities, position, or duties (0 = yes, 1 = no). The four scores constituted a work (in)stability index ranging from 0 to 4. The NFR's sensitivity to detect the effect of the increase in working hours was assessed indirectly by comparing it with an alternative scale, namely the Checklist Individual Strength. Test-retest reliability over a two year interval was good to excellent when applied in stable work environments (ICCs 0.68 to 0.80) but, as expected, poor to fair when applied in unstable work environments (ICCs 0.30 to 0.55). The NFR was sensitive in detecting an increase in work related fatigue due to the increase in working hours (effect size 0.40). The NFR's test-retest reliability and sensitivity to detect change are favourable. This implicates that the NFR may form a valuable part of health surveys and may be a useful tool for evaluating occupational healthcare interventions.


Portable and low-cost hologram verification module using a snapshot-based hyperspectral imaging algorithm.
One of the challenges in differentiating a duplicate hologram from an original one is reflectivity. A slight change in lighting condition will completely change the reflection pattern exhibited by a hologram, and consequently, a standardized duplicate hologram detector has not yet been created. In this study, a portable and low-cost snapshot hyperspectral imaging (HSI) algorithm-based housing module for differentiating between original and duplicate holograms was proposed. The module consisted of a Raspberry Pi 4 processor, a Raspberry Pi camera, a display, and a light-emitting diode lighting system with a dimmer. A visible HSI algorithm that could convert an RGB image captured by the Raspberry Pi camera into a hyperspectral image was established. A specific region of interest was selected from the spectral image and mean gray value (MGV) and reflectivity were measured. Results suggested that shorter wavelengths are the most suitable for differentiating holograms when using MGV as the parameter for classification, while longer wavelengths are the most suitable when using reflectivity. The key features of this design include low cost, simplicity, lack of moving parts, and no requirement for an additional decoding key.


Corticosteroids in Immunosuppression.
Corticosteroids have been utilized as mainstay pharmacological intervention for successful organ transplantation since the beginning. Several challenges exist in establishing a balance between achieving a tolerant atmosphere in the host immune system while minimizing the long-term impact of steroids on the body. Corticosteroids are used early in all solid organ transplantation but there is wide variability across various organs and centers in the duration of use and protocols of planned steroid wean. The adverse event profile of steroids is exhaustive and across many organ systems.


Partial characterization of T cell components related to defined VH (VT) markers.
Certain antigen-binding surface molecules and factors of T cells possess serological determinants related to immunoglobulin (Ig)-heavy-chain-variable regions (VH). We obtained sufficient quantities (greater than 100 micrograms) of homogenous VH-related T-cell molecules (VTM) for biochemical studies from normal murine thymocytes and by growing large quantities of monoclonal T-cell leukemia lines expressing the determinants. A solid phase immune adsorbent prepared from the IgG fraction of rabbit anti-IgT serum was used to isolate VTM from formic acid-solubilized T cells. The VTM from murine thymocytes and T-cell lines had Mr of 65,000-68,000. The VTM from distinct cell lines differ by isoelectric focusing and resolution of tryptic peptides indicating clonal restriction. VTM lack conventional light- or heavy-chain-constant region determinants but cross-react with antisera directed against defined VHa allotypes and JH peptides. The detection of a cross-reaction with a synthetic JH peptide is consistent with recently published data identifying JH-related sequences in putative T-cell receptor genes. The amino acid compositions of the VTM were distinct from those of mammalian Ig, major histocompatibility complex (MHC) antigens, and viral glycoproteins, but significant similarities occur with Ig V regions or heavy chains of primitive vertebrates. The results indicate that the VH-bearing T-cell products are not classical Ig, but bear limited VH-cross-reactive determinants.


Clinical, genetic, and histologic features of the trichoonychodental (TOD) syndrome.
A syndrome of scanty, fine, curled hair, thin dysplastic nails, taurodontic molars, hypoplastic-hypomature enamel, dysplasia of dentin, and hypohidrosis segregating as an autosomal dominant trait is described in a Japanese family. The patients received repeated doses of tetracycline during odontogenesis which fortuitously assisted in interpreting the pathogenesis of the dentin defect. The condition was previously described as familial congenital ectodermal dysplasia by Westerholm and as odontogenesis imperfecta by Siirilä and Heikinheimo. It is suggested that the name trichoonychodental (TOD) syndrome is a better descriptive term for the condition.


Surface electrostatics of lipid bilayers by EPR of a pH-sensitive spin-labeled lipid.
Many biophysical processes such as insertion of proteins into membranes and membrane fusion are governed by bilayer electrostatic potential. At the time of this writing, the arsenal of biophysical methods for such measurements is limited to a few techniques. Here we describe a, to our knowledge, new spin-probe electron paramagnetic resonance (EPR) approach for assessing the electrostatic surface potential of lipid bilayers that is based on a recently synthesized EPR probe (IMTSL-PTE) containing a reversibly ionizable nitroxide tag attached to the lipids' polar headgroup. EPR spectra of the probe directly report on its ionization state and, therefore, on electrostatic potential through changes in nitroxide magnetic parameters and the degree of rotational averaging. Further, the lipid nature of the probe provides its full integration into lipid bilayers. Tethering the nitroxide moiety directly to the lipid polar headgroup defines the location of the measured potential with respect to the lipid bilayer interface. Electrostatic surface potentials measured by EPR of IMTSL-PTE show a remarkable (within ±2%) agreement with the Gouy-Chapman theory for anionic DMPG bilayers in fluid (48°C) phase at low electrolyte concentration (50 mM) and in gel (17°C) phase at 150-mM electrolyte concentration. This agreement begins to diminish for DMPG vesicles in gel phase (17°C) upon varying electrolyte concentration and fluid phase bilayers formed from DMPG/DMPC and POPG/POPC mixtures. Possible reasons for such deviations, as well as the proper choice of an electrostatically neutral reference interface, have been discussed. Described EPR method is expected to be fully applicable to more-complex models of cellular membranes.


Smartphone Global Positioning System (GPS) Data Enhances Recovery Assessment After Breast Cancer Surgery.
We sought to determine whether smartphone GPS data uncovered differences in recovery after breast-conserving surgery (BCS) and mastectomy, and how these data aligned with self-reported quality of life (QoL). In a prospective pilot study, adult smartphone-owners undergoing breast surgery downloaded an application that continuously collected smartphone GPS data for 1 week preoperatively and 6 months postoperatively. QoL was assessed with the Short-Form-36 (SF36) via smartphone delivery preoperatively and 4 and 12 weeks postoperatively. Endpoints were trends in daily GPS-derived distance traveled and home time, as well as SF36 Physical (PCS) and Mental Component Scores (MCS) comparing BCS and mastectomy patients. Thirty-one patients were included. Sixteen BCS and fifteen mastectomy patients were followed for a mean of 201 (SD 161) and 174 (107) days, respectively. There were no baseline differences in demographics, PCS/MCS, home time, or distance traveled. Through 12 weeks postoperatively, mastectomy patients spent more time at home [e.g., week 4: 16.7 h 95% CI (14.3, 19.6) vs. 11.0 h (9.4, 12.9), p < 0.001] and traveled shorter distances [e.g., week 4: 52.5 km 95% CI (36.1, 76.0) vs. 107.7 km (75.8-152.9), p = 0.009] compared with BCS patients. There were no significant QoL differences throughout the study as measured by the MCS [e.g., week 4 difference: 7.83 95% CI (- 9.02, 24.7), p = 0.362] or PCS [e.g., week 4 difference: 8.14 (- 6.67, 22.9), p = 0.281]. GPS and QoL trends were uncorrelated (ρ < ± 0.26, p > 0.05). Differences in BCS and mastectomy recovery were successfully captured using smartphone GPS data. These data may describe currently unmeasured aspects of physical and mental recovery, which could supplement traditional and QoL outcomes to inform shared decision-making.


Epidemiology of nontuberculous mycobacteria in patients without HIV infection, New York City.
We reviewed medical records of patients without known HIV and with positive cultures for nontuberculous mycobacteria (NTM) isolated during 2000-2003 from 1 large hospital in New York, New York. Overall, 505 patients had positive NTM cultures; 119 (24%) met the criteria for NTM disease. The difference between demographic characteristics of case-patients in our study (66% female, 61% white, and 59% > 60 years of age) and those of the base population as determined by regional census data was statistically significant. Estimated incidences for positive cultures, all disease, and respiratory tract disease were 17.7, 2.7, and 2.0 per 100,000 persons, respectively. More patients with rapidly growing mycobacteria (61%), Mycobacterium kansasii (70%), or M. marinum (100%) met criteria for disease than did patients with M. avium complex (MAC) (27%, (p < 0.01). NTM disease in patients without HIV is increasing. Laboratory-based surveillance may be useful for detecting non-MAC and non-respiratory tract disease.


Limitations to the usefulness of N-succinimidyl propionate for the study of retrograde axonal transport.
Retrograde axonal transport of radiolabelled proteins was studied in rat sciatic nerve, after direct application of [3H]N-succinimidyl propionate. Waves of radiolabelled proteins were observed but only two proteins were predominantly labelled, one of molecular weight 68 kilodaltons (K) and the other of 19K. There was no evidence to confirm the waves as representing retrograde axonal transport of identifiable proteins, and the tendency of the covalent label to bind selectively in vivo to a small number of prominent proteins limits its usefulness for the detection of retrogradely transported proteins in general.


Implications of ACC/AHA Versus ESC/EAS LDL-C Recommendations for Residual Risk Reduction in ASCVD: A Simulation Study From DA VINCI.
Low-density lipoprotein cholesterol (LDL-C) recommendations differ between the 2018 American College of Cardiology/American Heart Association (ACC/AHA) and 2019 European Society of Cardiology/European Atherosclerosis Society (ESC/EAS) guidelines for patients with atherosclerotic cardiovascular disease (ASCVD) (< 70 vs. < 55 mg/dl, respectively). In the DA VINCI study, residual cardiovascular risk was predicted in ASCVD patients. The extent to which relative and absolute risk might be lowered by achieving ACC/AHA versus ESC/EAS LDL-C recommended approaches was simulated. DA VINCI was a cross-sectional observational study of patients prescribed lipid-lowering therapy (LLT) across 18 European countries. Ten-year cardiovascular risk (CVR) was predicted among ASCVD patients receiving stabilized LLT. For patients with LDL-C ≥ 70 mg/dl, the absolute LDL-C reduction required to achieve an LDL-C of < 70 or < 55 mg/dl (LDL-C of 69 or 54 mg/dl, respectively) was calculated. Relative and absolute risk reductions (RRRs and ARRs) were simulated. Of the 2039 patients, 61% did not achieve LDL-C < 70 mg/dl. For patients with LDL-C ≥ 70 mg/dl, median (interquartile range) baseline LDL-C and 10-year CVR were 93 (81-115) mg/dl and 32% (25-43%), respectively. Median LDL-C reductions of 24 (12-46) and 39 (27-91) mg/dl were needed to achieve an LDL-C of 69 and 54 mg/dl, respectively. Attaining ACC/AHA or ESC/EAS goals resulted in simulated RRRs of 14% (7-25%) and 22% (15-32%), respectively, and ARRs of 4% (2-7%) and 6% (4-9%), respectively. In ASCVD patients, achieving ESC/EAS LDL-C goals could result in a 2% additional ARR over 10 years versus the ACC/AHA approach.


Hepatic heat shock proteins, antioxidant-related genes, and immunocompetence of heat-stressed broilers in response to short periods of incubation during egg storage and thermal conditioning.
Short Periods of Incubation During Egg Storage (SPIDES) approach improves chick quality and hatching rates. Also, embryonic thermal conditioning (TC) is a strategy for enhancing thermotolerance in avian species. Until now, evaluating the effect of either SPIDES or embryonic TC effects has only been separately conducted, so we hypothesized that combining TC and SPIDES may enhance the response of broilers to thermal stress. Eight hundred Ross broiler eggs were divided into two groups; the first one was kept under appropriate storage room conditions, S0 (control) The 2nd was subjected to SPIDES for 5 h at 37.8 ○C ± 0.1 three times at days 5, 10, and 15 (S1) after egg collection respectively. On the 14th day of incubation (DOI) each of the two main groups was randomly divided into two equal subgroups; the control one was left under the appropriate incubation settings (TC0) whereas the other received prenatal heat conditioning (TC1) at 39.5 ○C ± 0.1 for 6 h/d from the 14th to the 18th embryonic day (E), resulting finally in four experimental subgroups (S0TC0, S1TC0, S0TC1 & S1TC1). RESULTS: showed that SPIDES treatment improved the hatchability of the stored eggs by almost 20% compared to untreated eggs. A combination of SPIDES and TC (S1TC1) increased significantly the levels of Immunoglobulin (IgG and IgM) production at hatch and heat-stressed birds. Our findings revealed that the hepatic heat shock proteins (hsp70, 90 A,90 B, 60 and hspA9), antioxidants-related genes (CAT, and SOD2), and NADPH4 were significantly downregulated in the thermally conditioned group that challenged with thermal stress conditions. As opposed to that, the SPIDES group showed a significant increase in hepatic heat shock proteins, antioxidants-related genes, and NADPH4 when subjected to thermal-stress conditions. In conclusion, the combination of SPIDES and TC has a positive effect on some pre and post-hatch traits of broiler chicks. Under heat stress challenge, thermal conditioning can modify the expression of antioxidant-related genes and Hsps, leading to the enhanced acquisition of thermotolerance as evidenced by lower expression of Hsps and NADPH4. While SPIDES does not have a significant role in thermotolerance acquisition.


Culture-free detection of methicillin-resistant Staphylococcus aureus by using self-driving diffusometric DNA nanosensors.
The rapid and robust detection of infectious bacteria is vital in sepsis treatment because of their ability to reveal multi-drug resistance. This study presents culture-free and self-driving DNA nanosensors by combining diffusometry and oligonucleotide probes to rapidly detect a lethal superbug, methicillin-resistant Staphylococcus aureus (MRSA). The DNA nanosensors were synthesized with conjugated fluorescent nanobeads and designed oligonucleotide probes that can recognize the target sequences on MRSA's genomic DNA. The high selectivity and specificity of this binding ensure the accuracy of detection. A DNA fragment tagged with gold nanoparticles (AuNPs) was attached to the same MRSA single-stranded DNA (ssDNA) to form a sandwiched configuration. The protrusive AuNPs surrounding the nanobeads decreased the diffusivity of the complexed nanobeads by increasing the bead size. Accordingly, diffusivity was inversely proportional to the concentration of the target MRSA ssDNA. Each measurement required only 10 s. An optimal limit of detection of 10 pM was achieved. This study successfully developed DNA nanosensors based on diffusometry for the rapid and robust detection of target superbugs and unknown pathogenic microorganisms.


Protein kinase C isoform ε negatively regulates ADP-induced calcium mobilization and thromboxane generation in platelets.
Members of the protein kinase C (PKC) family are shown to positively and negatively regulate platelet activation. Although positive regulatory roles are extensively studied, negative regulatory roles of PKCs are poorly understood. We investigated the mechanism and specific isoforms involved in PKC-mediated negative regulation of ADP-induced functional responses. A pan-PKC inhibitor, GF109203X, potentiated ADP-induced cPLA(2) phosphorylation and thromboxane generation as well as ERK activation and intracellular calcium (Ca(2+)(i)) mobilization, 2 signaling molecules, upstream of cPLA(2) activation. Thus, PKCs inhibit cPLA(2) activation by inhibiting ERK and Ca(2+)(i) mobilization. Because the inhibitor of classic PKC isoforms, GO-6976, did not affect ADP-mediated thromboxane generation, we investigated the role of novel class of PKC isoforms. ADP-induced thromboxane generation, calcium mobilization, and ERK phosphorylation were potentiated in PKCε null murine platelets compared with platelets from wild-type littermates. Interestingly, when thromboxane release is blocked, ADP-induced aggregation in PKCε knockout and wild-type was similar, suggesting that PKCε does not affect ADP-induced aggregation directly. PKCε knockout mice exhibited shorter times to occlusion in an FeCl(3)-induced arterial injury model and shorter bleeding times in tail-bleeding experiments. We conclude that PKCε negatively regulates ADP-induced thromboxane generation in platelets and offers protection against thrombosis.


Understanding the cometabolic degradation of sulfadiazine by an enriched ammonia oxidizing bacteria culture from both extracellular and intracellular perspectives.
Antibiotics are widely used drugs in the world and pose serious threats to ecosystems and human health. Although it has been reported that ammonia oxidizing bacteria (AOB) can cometabolize antibiotics, little has been reported on how AOB would respond to the exposure of antibiotics on extracellular and enzymatic levels, as well as the impact of antibiotics on the bioactivity of AOB. Therefore, in this study, a typical antibiotic, sulfadiazine (SDZ), was selected, and a series short-term batch tests using enriched AOB sludge were conducted to investigate the intracellular and extracellular responses of AOB along the cometabolic degradation process of SDZ. The results showed the cometabolic degradation of AOB made the main contribution to SDZ removal. When the enriched AOB sludge was exposed to SDZ, ammonium oxidation rate, ammonia monooxygenase activity, adenosine triphosphate concentration and dehydrogenases activity were negatively affected. The amoA gene abundance increased 1.5 folds within 24 h, which may enhance the uptake and utilization of substrates and maintain stable metabolic activity. In the tests with and without ammonium, the concentration of total EPS increased from 264.9 to 231.1 mg/gVSS to 607.7 and 538.2 mg/gVSS, respectively, under the exposure to SDZ, which was mainly contributed by the increase of proteins in tightly bound extracellular polymeric substances (EPS) and polysacharides in tightly bound EPS and soluble microbial products. The proportion of tryptophan-like protein and humic acid-like organics in EPS also increased. Moreover, SDZ stress stimulated the secretion of three quorum sensing signal molecules, C4-HSL (from 140.3 to 164.9 ng/L), 3OC6-HSL (from 17.8 to 42.4 ng/L) and C8-HSL (from 35.8 to 95.9 ng/L) in the enriched AOB sludge. Among them, C8-HSL may be a key signal molecule that promoted the secretion of EPS. The findings of this study could shed more light on the cometabolic degradation of antibiotics by AOB.


Kleptoplasty: Getting away with stolen chloroplasts.
Kleptoplasty, the process by which a host organism sequesters and retains algal chloroplasts, is relatively common in protists. The origin of the plastid varies, as do the length of time it is retained in the host and the functionality of the association. In metazoa, the capacity for long-term (several weeks to months) maintenance of photosynthetically active chloroplasts is a unique characteristic of a handful of sacoglossan sea slugs. This capability has earned these slugs the epithets "crawling leaves" and "solar-powered sea slugs." This Unsolved Mystery explores the basis of chloroplast maintenance and function and attempts to clarify contradictory results in the published literature. We address some of the mysteries of this remarkable association. Why are functional chloroplasts retained? And how is the function of stolen chloroplasts maintained without the support of the algal nucleus?


Dynamics of phytoplankton productivity and exopolysaccharides (EPS and TEP) pools in the Seine Estuary (France, Normandy) over tidal cycles and over two contrasting seasons.
Exopolysaccharides (EPS) play an important role in the carbon flux and may be directly linked to phytoplankton and microphytobenthos production, most notably in estuarine systems. However the temporal and spatial dynamics of estuarine EPS are still not well understood, nor how primary productivity triggers this variability at these different scales. The aim of this study was to investigate the primary productivity of phytoplankton and EPS dynamics in the Seine estuary over a tidal cycle in three different haline zones over two contrasted seasons. The other objectives was to investigate the origin of pools of soluble carbohydrates (S-EPS) and transparent exopolymeric particles (TEP) in phytoplankton, microphytobenthos or other compartments. High frequency measurements of productivity were made in winter and summer 2015. Physical and chemical parameters, biomass and EPS were measured at hourly intervals in sub-surface waters and just above the water sediment-interface. Our results confirmed that high frequency measurements improve the accuracy of primary productivity estimations and associated carbon fluxes in estuaries. The photosynthetic parameters were shown to be strongly controlled by salinity and by the concentrations of suspended particle matter at the smallest temporal and at spatial scales. At these scales, our results showed an inverse relationship between EPS concentrations and biomass and productivity, and a positive relationship with sediment resuspension. Additionally, the distribution of EPS appears to be linked to hydrodynamics with the tide at daily scale and with the winter at seasonal scale. At spatial scale, the maximum turbidity zone played an important role in the distribution of TEP. Our results suggest that, in the Seine estuary, between 9% and 33% of the S-EPS pool in the water column can be attributed to phytoplankton excretion, while only 0.4%-1.6% (up to 6.14% in exceptional conditions) originates from the microphytobenthos compartments. Most EPS was attributed to remobilization of detrital carbon pools in the maximum turbidity zone and in the sediment or allochthonous origin.


Elementary School-Aged Children in Therapeutic Residential Care: Examining Latent Classes, Service Provision, and Outcomes.
Approximately one-third of children in residential care are elementary-school aged. Yet, little is known about the subset of younger children in residential care and the nature of these placements. This study identified latent classes of younger children in residential care and compared the purposes for placement, treatment processes, and outcomes across classes. The sample included 216 children (ages 5-10) placed in therapeutic residential care. A three-step latent class model was used to estimate conditional effects of class membership on impairment at discharge, length of stay, and discharge placement. A content analysis of a randomly selected sample of case records from each class was used to explore placement processes. There were three classes identified (class 1: child welfare/multi-problem families; class 2: mental-health/angry-oppositional; class 3: strong families/attachment). All classes experienced large improvements in functioning. Children in class 3 were in care longer (CI95% 1.72, 15.48) and experienced greater reductions in impairment (CI95% -11.17, -32.06) than class 2. Classes did not differ in rates of discharge to family-based care, however, more children in classes 1 (20.9%) and 3 (21.6%) discharged to group-based placements than class 2 (11.1%). The content analysis revealed similarities in reasons for placement and treatment processes across classes with some distinctions. Service goals were similar across classes and focused on emotional management, social skills, and developing trust. The results supported individualized approaches to facilitate discharge to stable, family-based care and reduced risks for re-entry and prolonged out-of-home care for younger children.


Demonstration of immunoglobulin in brains of ferrets inoculated with an SSPE strain of measles virus: use of protein A conjugated to horseradish peroxidase.
Measles virus-specific immunoglobulin G (IgG) has been found in the brains of patients with subacute sclerosing panencephalitis (SSPE), a slowly progressing central nervous system (CNS) disease affecting children. IgG/albumin ratios indicate that the antibodies are probably synthesized in the CNS. In a ferret model system, protein A conjugated to horseradish peroxidase (PrAPx) was used to localize Ig's in brains of animals inoculated with a cell associated strain of SSPE. Ig's were found in plasma cells in various stages of antibody production both in perivascular inflammatory lesions and scattered throughout the cerebral cortex. These findings offer corroborative evidence that the Ig found in SSPE ferret brain and CSF is actively synthesized within the CNS. Antibody was also demonstrated in glial and neuronal cell bodies and processes and in postsynaptic profiles. These are the same sites where measles virus antigens are most frequently found and suggests the possibility of immune complex formation.


Influence of amino acids on hybridoma cell viability and antibody secretion.
It is generally accepted that the phase of cell decline observed in batch culture of mammalian cells is related to exhaustion of medium nutrients (principally glucose and glutamine) and/or to waste products accumulation. In the present paper, we have studied the influence of glutamine on the proliferation of mouse hybridoma cells. We showed that repeated addition of glutamine prolonged the life span of the culture and significantly increased the secretion of monoclonal antibody. Flow cytometry analysis suggests that this effect of glutamine is related to a delay in cell death rather than to a stimulation of proliferation.Addition of glutamine and glucose failed however to prevent the death of the culture. Determinations of amino acid consumption in glutamine-supplemented samples and experiments carried out with complementary sources of amino acids (e.g. tryptose phosphate) strongly suggest that amino acid supply is a critical factor governing cell growth and productivity.


Fibrous soft tissue tumors: factors predictive of recurrence.
Fibrous soft tissue tumors (FSTT) of infancy and childhood present a challenge to surgeons due to ambiguous histological interpretation, propensity for local recurrence, and uncertainty regarding appropriate therapy. Records were reviewed for patients diagnosed and treated for FSTT at our institution from 1992 to 2010. Data included demographics, tumor location, operative treatment, histopathology, margin status, mitotic index, and recurrence. Eighty-six patients underwent treatment of FSTT. Mean follow-up time was 62 months (range 1 to 201 months). There were 11 recurrences (13%), with a mean time to recurrence of 23 months. Extremity and digital lesions were most likely to recur with 21% of such lesions recurring compared to 7% of lesions located elsewhere. Recurrences were seen more frequently in older children. Despite the presence of positive margins in 92% of cases, margin status, histopathology, and mitotic index were not significant predictors of recurrence. Digital and extremity fibrous soft tissue tumors and tumors of older children have an increased risk for local recurrence. Margin status did not predict recurrence rates in this series. While the surgical goal is to obtain complete local resection with clear margins, this may be challenging in particular locations and may not ultimately impact recurrence.


Variants in FGF10 cause early onset of severe childhood interstitial lung disease: A detailed description of four affected children.
Fibroblast growth factor 10 (FGF10) is a signaling molecule with a well-established role for lung branching morphogenesis. Rare heterozygous, deleterious variants in the FGF10 gene are known causes of the lacrimo-auriculo-dento-digital (LADD) syndrome and aplasia of lacrimal and salivary glands. Previous studies indicate that pathogenic variants in FGF10 can cause childhood Interstitial Lung Disease (chILD) due to severe diffuse developmental disorders of the lung, but detailed reports on clinical presentation and follow-up of affected children are lacking. We describe four children with postnatal onset of chILD and heterozygous variants in FGF10, each detected by exome or whole genome sequencing. All children presented with postnatal respiratory failure. Two children died within the first 2 days of life, one patient died at age of 12 years due to right heart failure related to severe pulmonary hypertension (PH) and one patient is alive at age of 6 years, but still symptomatic. Histopathological analysis of lung biopsies from the two children with early postpartum demise revealed diffuse developmental disorder representing acinar dysplasia and interstitial fibrosis. Sequential biopsies of the child with survival until the age of 12 years revealed alveolar simplification and progressive interstitial fibrosis. Our report extends the phenotype of FGF10-related disorders to early onset chILD with progressive interstitial lung fibrosis and PH. Therefore, FGF10-related disorder should be considered even without previously described syndromic stigmata in children with postnatal respiratory distress, not only when leading to death in the neonatal period but also in case of persistent respiratory complaints and PH.


Sustained replication of dengue pseudoinfectious virus lacking the capsid gene by trans-complementation in capsid-producing mosquito cells.
A simple system for the generation of pseudoinfectious particles of dengue virus was developed to facilitate studies of virus replication and vaccine development. Selected clones of the C6/36 mosquito cell line expressing an anchored form of the dengue virus capsid protein served as host cells for the trans-complementation of partially capsid-deleted viral RNA generated in vitro. Transfection of the partially capsid-deleted viral RNA into the anchored capsid-expressing C6/36 cells resulted in moderate titers of infectious virus. Progeny viruses multiplied in the capsid trans-complementing C6/36 cells for up to three weeks, but only initiated single rounds of replication in Vero cells lacking the capsid protein. Employing this trans-complementation system, it was found that nearly all of the capsid-coding sequence in the viral RNA was dispensable for the generation of pseudoinfectious dengue virus particles in mosquito cells.


Comparison of the effects of serotonin in the hippocampus and the entorhinal cortex.
Among the molecular, cellular, and systemic events that have been proposed to modulate the function of the hippocampus and the entorhinal cortex (EC), one of the most frequently cited possibilities is the activation of the serotonergic system. Neurons in the hippocampus and in the EC receive a strong serotonergic projection from the raphe nuclei and express serotonin (5-HT) receptors at high density. Here we review the various effects of 5-HT on intrinsic and synaptic properties of neurons in the hippocampus and the EC. Although similar membrane-potential changes following 5-HT application have been reported for neurons of the entorhinal cortex and the hippocampus, the effects of serotonin on synaptic transmission are contrary in both areas. Serotonin mainly depresses fast and slow inhibition of the principal output cells of the hippocampus, whereas it selectively suppresses the excitation in the entorhinal cortex. On the basis of these data, we discuss the possible role of serotonin under physiological and pathophysiological circumstances.


Fractional seroprevalence rates in common prey species can cause more than half of feral cats to be exposed to Toxoplasma gondii annually.
Rodents comprise a major component of cat (Felis catus) diets in many ecosystems, and life cycle diagrams of Toxoplasma gondii typically depict small rodents as quintessential intermediate hosts. Counter-intuitively, small rodents often experience a lower T. gondii seroprevalence than do larger sympatric mammals. This observation has repeatedly caused confusion about the relative importance of small rodents to the ecology of T. gondii. To address this confusion, we modified the Reed-Frost epidemic model to develop a simple binomial equation to model T. gondii transmission from prey to feline predators. This equation takes into account variations in prey seroprevalence and the frequency with which they are consumed by felids. Even when T. gondii seroprevalence in prey is < 1%, computation reveals that the risk of feline exposure to T. gondii can easily exceed 50 % annually. For example, if cats eat an average of 1 mouse per day, a seroprevalence of 0.2 % (1/500) in mice will cause 51.9 % of cats to be exposed to T. gondii annually. Our simple equation demonstrates that both prey seroprevalence and the rate at which prey are consumed are of approximately equal importance to the ecology of T. gondii. When inferring the importance of various prey species to the ecology of T. gondii, researchers must consider the predation and dietary habits of felids from within their study system. Our simple binomial equation could also be used to predict T. gondii exposure rates of humans or other carnivorous animals from various dietary sources or be applied to other predator-prey parasite life cycles.


Analysis of coenzyme A activated compounds in actinomycetes.
Acyl-CoAs are crucial compounds involved in essential metabolic pathways such as the Krebs cycle and lipid, carbohydrate, and amino acid metabolisms, and they are also key signal molecules involved in the transcriptional regulation of lipid biosynthesis in many organisms. In this study, we took advantage of the high selectivity of mass spectrometry and developed an ion-pairing reverse-phase high-pressure liquid chromatography electrospray ionization high-resolution mass spectrometry (IP-RP-HPLC/ESI-HRMS) method to carry on a comprehensive analytical determination of the wide range of fatty acyl-CoAs present in actinomycetes. The advantage of using a QTOF spectrometer resides in the excellent mass accuracy over a wide dynamic range and measurements of the true isotope pattern that can be used for molecular formula elucidation of unknown analytes. As a proof of concept, we used this assay to determine the composition of the fatty acyl-CoA pools in Mycobacterium, Streptomyces, and Corynebacterium species, revealing an extraordinary difference in fatty acyl-CoA amounts and species distribution between the three genera and between the two species of mycobacteria analyzed, including the presence of different chain-length carboxy-acyl-CoAs, key substrates of mycolic acid biosynthesis. The method was also used to analyze the impact of two fatty acid synthase inhibitors on the acyl-CoA profile of Mycobacterium smegmatis, which showed some unexpected low levels of C24 acyl-CoAs in the isoniazid-treated cells. This robust, sensitive, and reliable method should be broadly applicable in the studies of the wide range of bacteria metabolisms in which acyl-CoA molecules participate.


Functional characterization and development of novel human kinase insert domain receptor chimeric antigen receptor T-cells for immunotherapy of non-small cell lung cancer.
CAR-T cell therapy, in which T cells are transfected or transduced with a chimeric antigen receptor (CAR), is a transformative type of cancer immunotherapy. Despite outstanding success in hematological malignancies, their efficacy against solid tumors has been limited. Here, we aimed to explore whether T cells modified by a CAR targeting the vascular endothelial growth factor 2 receptor/ kinase insert domain receptor (KDR) could destroy tumors and their vasculature. A second-generation KDR-CAR was constructed and transfected into T cells using lentivirus. The 3D structure of the CAR construct and target antigen was predicted. Moreover, in silico analysis, including molecular docking and molecular dynamics (MD) simulation, were used to evaluate the minimum energy of interaction and stability of the complex. The anti-cancer effect of KDR-specific CAR-T cells was tested with KDR-expressing and KDR overexpressing A549 cell line. The in-silico study suggested that this CAR construct could be effective for lung cancer therapy. We evaluated this using both in vitro and in vivo experiments. The KDR-CAR-T cells targeted and killed KDR-A549 with high efficiency by expressing IFN-γ and releasing granzyme B. The in vivo study showed that KDR-CAR-T cells dramatically inhibited the growth of lung cancer KDR-A549 xenografts in BALB/c-nu mice at day 10. The characterization of T cells modified by KDR-CAR by computational biology and wet-lab experiments suggested its applicability as a new treatment strategy for lung cancer and, potentially, for other vascularized solid tumors.


The CombinADO study to assess the impact of a combination intervention strategy on viral suppression, antiretroviral therapy adherence, and retention in HIV care among adolescents and young people living with HIV: protocol for a cluster-randomized controlled trial.
Adolescents and youth living with HIV (AYAHIV) have worse HIV outcomes than other age groups, particularly in sub-Saharan Africa (SSA). AYAHIV in SSA face formidable health system, interpersonal- and individual-level barriers to retention in HIV care, uptake of ART, and achievement of viral suppression (VS), underscoring an urgent need for multi-component interventions to address these challenges. This cluster-randomized control trial (cRCT) aims to evaluate the effectiveness and monitor implementation of a community-informed multi-component intervention ("CombinADO strategy") addressing individual-, facility-, and community-level factors to improve health outcomes for AYAHIV. This trial will be conducted in 12 clinics in Nampula Province, Northern Mozambique. All clinics will implement an optimized standard of care (control) including (1) billboards/posters and radio shows, (2) healthcare worker (HCW) training, (3) one-stop adolescent and youth-friendly services, (4) information/motivation walls, (5) pill containers, and (6) tools to be used by HCW during clinical visits. The CombinADO strategy (intervention) will be superadded to control conditions at 6 randomly selected clinics. It will include five additional components: (1) peer support, (2) informational/motivational video, (3) support groups for AYAHIV caregivers, (4) AYAHIV support groups, and (5) mental health screening and linkage to adolescent-focused mental health support. The study conditions will be in place for 12 months; all AYAHIV (ages 10-24 years, on ART) seeking care in the participating sites will be exposed to either the control or intervention condition based on the clinic they attend. The primary outcome is VS (viral load < 50 copies/mL) at 12 months among AYAHIV attending participating clinics. Secondary outcomes include ART adherence (self-reported and TDF levels) and retention in care (engagement in the preceding 90 days). Uptake, feasibility, acceptability, and fidelity of the CombinADO strategy during implementation will be measured. Trial outcomes will be assessed in AYAHIV, caregivers, healthcare workers, and key informants. Statistical analyses will be conducted and reported in line with CONSORT guidelines for cRCTs. The CombinADO study will provide evidence on effectiveness and inform implementation of a novel community-informed multi-component intervention to improve retention, adherence, and VS among AYAHIV. If found effective, results will strengthen the rationale for scale up in SSA. ClinicalTrials.gov NCT04930367 . Registered on 18 June 2021.


15-deoxy-Δ12,14 -prostaglandin J2 reduces recruitment of bone marrow-derived monocyte/macrophages in chronic liver injury in mice.
15-Deoxy-Δ(12,14) -Prostaglandin J(2) (15d-PGJ(2) ), a natural peroxisome proliferator-activated receptor gamma (PPAR-γ) ligand, has been implicated as a new antiinflammatory compound with possible clinical applications. Based on this concept, this study was designed to evaluate the effects of 15d-PGJ(2) on bone marrow-derived monocyte/macrophage (BMM) migration, phagocytosis, and cytokine expression after liver injury using mouse models induced by cholestasis or carbon tetrachloride. Mice were lethally irradiated and received bone marrow transplants from enhanced green fluorescent protein transgenic mice. Our results showed that recruitment of BMM was significantly increased during chronic liver injury, and that 15d-PGJ(2) administration reduced BMM, but not neutrophil, dendritic, or T cell migration toward the damaged liver, involving reactive oxygen species generation and independently of PPAR-γ. Moreover, 15d-PGJ(2) inhibited the phagocytic activity of BMM and down-regulated inflammatory cytokine expression in vivo and in vitro. Accordingly, hepatic inflammation and fibrosis were strikingly ameliorated after 15d-PGJ(2) administration. Our findings strongly suggest the antiinflammation and antifibrogenic potential of 15d-PGJ(2) in chronic liver diseases.


Repetition, not number of sources, increases both susceptibility to misinformation and confidence in the accuracy of eyewitnesses.
Are claims more credible when made by multiple sources, or is it the repetition of claims that matters? Some research suggests that claims have more credibility when independent sources make them. Yet, other research suggests that simply repeating information makes it more accessible and encourages reliance on automatic processes-factors known to change people's judgments. In Experiment 1, people took part in a "misinformation" study: people first watched a video of a crime and later read eyewitness reports attributed to one or three different eyewitnesses who made misleading claims in either one report or repeated the same misleading claims across all three reports. In Experiment 2, people who had not seen any videos read those same reports and indicated how confident they were that each claim happened in the original event. People were more misled by-and more confident about-claims that were repeated, regardless of how many eyewitnesses made them. We hypothesize that people interpreted the familiarity of repeated claims as markers of accuracy. These findings fit with research showing that repeating information makes it seem more true, and highlight the power of a single repeated voice.


New HPLC method for separation of blood plasma phospholipids.
The aim of the present work was to develop a new HPLC method for separation of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI) and lysophosphatidylcholine (LPC) from small-volume samples of blood plasma. Human plasma glycerophospholipids were separated by liquid-liquid extraction method followed by solid phase extraction (SPE) on aminopropyl columns. Reversed-phase Sephasil C8 column (10 cm x 2.1 mm, I.D. 5 microm) and micropreparative chromatograph "SMART" were used for separation of PC, PE, LPC and PI from SPE phospholipids extract. Binary-step gradient of eluent A: acetonitrile-methanol (130:5, v/v) and B (0.01% trifluoroacetic acid) provided good, fast and reproducible resolution of investigated phospholipids classes in 12 min at 30 degrees C. Eluted phospholipids were detected at wavelengths lambda=235 and 254 nm. This method made it possible to determine quantitatively: 5 microg ml(-1) PC, 1 microg ml(-1) LPC, 4 microg ml(-1) PE and 3 microg ml(-1) PI in blood plasma samples.


Porcine bone grafts defatted by lipase: efficacy of defatting and assessment of cytocompatibility.
Defatting is an important procedure for the preparation of bone grafts because lipids in bone grafts strongly influence the osteointegration. Lipases have been widely used in different fields. However, study on the application to defatting process for bone grafts preparation has never been found so far. In this study, bone samples were treated respectively by lipase, NaHCO(3)/Na(2)CO(3), acetone and deionized water. The lipids content of processed bone grafts was calculated in Soxhlet extractor method. Surface morphology of the bone grafts was observed under scanning electron microscope (SEM). DNA content of processed bone grafts was measured. Cytocompatibility was evaluated by co-culturing mouse preosteoblasts (MC3T3-E1) on defatted bone cubes. Proliferation rates of MC3T3-E1 were examined by cell counting kit-8 (CCK-8) assay. No statistically significant difference was found between lipids amount of bone processed by lipase (0.46 ± 0.16 %) and acetone (1.11 ± 0.13 %) (P > 0.05). Both of them were significantly lower than that in groups processed by Na(2)CO(3)/NaHCO(3) (3.46 ± 0.69 %) and deionized water (8.88 ± 0.18 %) (P = 0.000). Only cell debris were discovered over the surface of bone processed by lipase or acetone, while lipid droplets were observed on bone processed by Na(2)CO(3)/NaHCO(3) or water by SEM. The difference of DNA concentration between the bone processed by lipase (3.16 ± 0.81 ng/μl) and acetone (4.14 ± 0.40 ng/μl) is not statistically significant (P > 0.05). Both of them are significantly lower than that groups processed by Na(2)CO(3)/NaHCO(3) (5.22 ± 0.38 ng/μl) and water (7.88 ± 0.55 ng/μl) (P < 0.05). MC3T3-E1 cells maintained their characteristic spreading on the trabecular surfaces of bone processed by lipase. There were no statistically significant differences among absorbance of lipase, acetone groups in CCK-8 assay. The application of lipase to bone tissue defatting appears to be a very promising technique for bone grafts preparation.


Multimodal fusion models for pulmonary embolism mortality prediction.
Pulmonary embolism (PE) is a common, life threatening cardiovascular emergency. Risk stratification is one of the core principles of acute PE management and determines the choice of diagnostic and therapeutic strategies. In routine clinical practice, clinicians rely on the patient's electronic health record (EHR) to provide a context for their medical imaging interpretation. Most deep learning models for radiology applications only consider pixel-value information without the clinical context. Only a few integrate both clinical and imaging data. In this work, we develop and compare multimodal fusion models that can utilize multimodal data by combining both volumetric pixel data and clinical patient data for automatic risk stratification of PE. Our best performing model is an intermediate fusion model that incorporates both bilinear attention and TabNet, and can be trained in an end-to-end manner. The results show that multimodality boosts performance by up to 14% with an area under the curve (AUC) of 0.96 for assessing PE severity, with a sensitivity of 90% and specificity of 94%, thus pointing to the value of using multimodal data to automatically assess PE severity.


Production of the kringle fragments of human apolipoprotein(a) by continuous lactose induction strategy.
A novel lactose induction strategy for the production of rhLK68, the kringle fragments of human apolipoprotein(a) (apo(a)) as a novel anti-angiogenic protein, was investigated. A scale-up of the production was accompanied by a decrease in expression level, and severe aggregation occurred during the solubilization of rhLK68 from the inclusion body during a conventional single introduction of lactose. To overcome this problem, a continuous induction strategy was applied where lactose was mixed with glycerol and fed continuously in a dissolved oxygen (DO)-stat manner. With the sub-optimal feed medium consisted of 1:50 of lactose/glycerol (w/w), the expression level reached 16% of the total cellular protein, which was 1.6-fold higher than that obtained from the conventional lactose induction. Moreover, the solubilization yield of rhLK68 from the inclusion body increased from 30 +/- 5 to 85 +/- 3% compared to the conventional single introduction of lactose. This result suggests that the continuous lactose induction strategy beneficially influenced the expression level of rhLK68 and the quality of its inclusion body.


Integrally calcified solitary fibrous tumor in the retroperitoneum: a case report and review of the literature.
Solitary fibrous tumor (SFT) is a rare stromal neoplasm and usually occurs in the thoracic cavity. We here report a case of retroperitoneal SFT with prominent calcification. A 64-year-old man presented with an incidentally detected retroperitoneal mass in the right upper abdomen. Imaging tests indicated an integrally calcified mass. The lesion was observed for 2 years and laparoscopically resected according to the patient's wish. Microscopically, the mass was mostly occupied by calcification and proliferous spindle cells were scattered with positive CD34 expression. We diagnosed morphologically benign SFT and the patient remained disease-free 1 year after the excision. There has been no report of such integrally calcified SFT. Retroperitoneal SFT is difficult to make a preoperative diagnosis, and careful follow-up after the excision is recommended because morphological malignancy does not always correspond to clinical malignancy.


The packaging of siRNA within the mesoporous structure of silica nanoparticles.
Mesoporous silica nanoparticle (MSN) is a promising material for biomedical applications, such as delivering drugs or biological molecules (siRNA or DNA), to the target cells or tissues. With positive-charge functionalization on their surface, MSNs have already been used as vectors for siRNA delivery. Nevertheless, such siRNA packaging strategy avoids utilizing the mesopores and consequently hinders further modifications on the delivery vehicle surface. To solve these problems, we have successfully packaged siRNA into the mesopores of magnetic mesoporous silica nanoparticles (M-MSNs) under a strongly dehydrated solution condition. The siRNA-loaded M-MSNs were mixed with polyethyleneimine (PEI) to form a polymer layer on their external surface. The obtained aggregates were further treated by ultrasonication in acidic solution to prepare well dispersed siRNA delivery vehicles (M-MSN_siRNA@PEI). Such delivery vehicles, with effective siRNA protective effect and negligible cytotoxicity, could be internalized into cancer cells and release siRNA in the cytoplasm. In gene silencing experiments, these delivery vehicles mediated, with high efficiency, knockdown of both exogenous enhanced green fluorescent protein (EGFP) gene and endogenous B-cell lymphoma 2 (Bcl-2) gene. In summary, our siRNA packaging strategy extends the application potential of M-MSNs and the resulting siRNA delivery vehicles can be further tested for in vivo experiments.


Endovascular coiling of small intracranial aneurysms using a very soft bare platinum coil: A comparison of the packing performance of new and old HyperSoft® helical coils.
Soft and small coils are known to be appropriate for the treatment of small aneurysms. The purpose of this study was to determine whether the new HyperSoft® helical coil, which is softer and smaller than its predecessors, has any effect on the packing performance in a matched-pair study with an old HyperSoft® helical coil. Thirty-six consecutive patients harboring 43 cerebral aneurysms treated with new HyperSofts were included in this study. Forty-one aneurysms treated with old HyperSofts were identified from our database as matched controls based on similar volumes and locations. Packing attenuation, adverse events during the procedures, and angiographic occlusions were observed and compared between the two groups. The mean packing density was significantly higher in the new HyperSoft® group compared to the control group (35.5% vs. 26.9%), and a larger proportion of the aneurysms embolized with the 1.5 mm size coil, which has higher packing density. There was no difference in immediate and midterm angiographic outcomes. There was no difference in the rate of intraprocedural perforation, but there was no intraprocedural rupture related to the 1.5 mm coil. The use of new HyperSoft® helical coils allows higher packing density comparable with the old technology. New HyperSoft® coils, especially those with 1.5 mm loop diameter, can be expected to fill smaller residual spaces in small aneurysms and may be helpful in preventing recanalization.


Solubility advantage of amorphous pharmaceuticals: II. Application of quantitative thermodynamic relationships for prediction of solubility enhancement in structurally diverse insoluble pharmaceuticals.
To quantitatively assess the solubility advantage of amorphous forms of nine insoluble drugs with a wide range of physico-chemical properties utilizing a previously reported thermodynamic approach. Thermal properties of amorphous and crystalline forms of drugs were measured using modulated differential calorimetry. Equilibrium moisture sorption uptake by amorphous drugs was measured by a gravimetric moisture sorption analyzer, and ionization constants were determined from the pH-solubility profiles. Solubilities of crystalline and amorphous forms of drugs were measured in de-ionized water at 25°C. Polarized microscopy was used to provide qualitative information about the crystallization of amorphous drug in solution during solubility measurement. For three out the nine compounds, the estimated solubility based on thermodynamic considerations was within two-fold of the experimental measurement. For one compound, estimated solubility enhancement was lower than experimental value, likely due to extensive ionization in solution and hence its sensitivity to error in pKa measurement. For the remaining five compounds, estimated solubility was about 4- to 53-fold higher than experimental results. In all cases where the theoretical solubility estimates were significantly higher, it was observed that the amorphous drug crystallized rapidly during the experimental determination of solubility, thus preventing an accurate experimental assessment of solubility advantage. It has been demonstrated that the theoretical approach does provide an accurate estimate of the maximum solubility enhancement by an amorphous drug relative to its crystalline form for structurally diverse insoluble drugs when recrystallization during dissolution is minimal.


Experimental spectroscopic (FTIR, FT-Raman, FT-NMR, UV-Visible) and DFT studies of 1-ethyl-1,4-dihydro-7-methyl-4oxo-1,8 napthyridine-3-carboxylic acids.
The solid phase FTIR and FT-Raman spectra of 1-ethyl-1,4-dihydro-7-methyl-4oxo-1,8 napthyridine-3-carboxylic acid (EDMONCA) have been recorded in the regions 4000-500 and 4000-400 cm(-1) respectively. The equilibrium geometry, harmonic vibrational frequencies have been investigated by DFT/B3LYP and B3PW91 methods with 6-311G (d,p) basis set. The different between the observed and scaled wave number values of most of the fundamental is very small. The assignments of the vibrational spectra have been carried out with the aid of normal coordinate analysis (NCA) following the scaled quantum mechanical force field methodology (SQMFFM). Stability of the molecule arising from hyper conjugative interactions, charge delocalization has been analyzed using natural bond orbital (NBO) analysis. UV-Visible spectrum of the compound was recorded and the electronic properties HOMO and LOMO energies were measured. The electric dipole moment (μD) and first hyperpolarizability (βtot) values of the investigated molecule were computed using ab initio quantum mechanical calculations. The calculated results also show that the EDMONCA molecule may have microscopic nonlinear optics (NLO) behavior with non-zero values. (1)H and (13)C NMR spectra were recorded and (1)H and (13)C nuclear magnetic resonance chemical shift of the molecule were calculated using the gauge independent atomic orbital (GIAO) method. Thermal stability of EDMONCA was studied by thermogravimetric analysis (TGA). Next Fukui function was calculated to explain the chemical selectivity or reactivity site in EDMONCA. Finally molecular electrostatic potential (MEP) and other molecular properties were performed.


Mental disorders in primary care: epidemiologic, diagnostic, and treatment research directions.
An extensive series of investigations over the past 2 decades clearly demonstrate that mental disorders are present in approximately 25% of primary care patients and that physicians underdiagnose these illnesses. The factors producing this bias are poorly understood and should be focused upon in future research. Also requiring much more study is the efficacy of pharmacologic and psychosocial treatments initially validated with psychiatric populations. Clinical trials should determine whether standardized interventions can be utilized with medical patients whose symptom profiles and organic comorbidity may differ from those of psychiatric populations.


Unveiling the treatment-driven secondary risks of pharmaceuticals: New lessons from calcium channel blocker transformation by multiple oxidants.
The worldwide detection of numerous pharmaceuticals and their transformation products (TPs) in different environmental matrices has gained considerable concern about their potential ecological hazards. Increasing evidence suggested that calcium channel blockers (CCBs) are ubiquitous pharmaceutical pollutants in natural waters. However, their TPs, reaction pathways, and secondary risks have been limitedly known during oxidative water treatment. This study systematically assessed the TP formation and transformation mechanisms of two typical CCBs (i.e., amlodipine, AML; verapamil, VER) oxidized by ferrate(VI), permanganate, and ozone, as well as the in silico prediction on the TPs' properties. The high-resolution mass spectrometer analysis suggested a total of 16 TPs of AML and 8 TPs of VER identified for these reaction systems. Transformation of AML mainly proceeded through hydroxylation of the aromatic ring, ether bond cleavage, NH2 substitution by a hydroxyl group, and H-abstraction, while VER was oxidized via hydroxylation/opening of the aromatic ring and cleavage of the CN bond. Notably, certain TPs of both CCBs were estimated with low biodegradation, multi-endpoint toxicity, and high persistence and bioaccumulation, suggesting their severe risks to aquatic ecosystems. This study has implications for understanding the environmental behaviors, fate, and secondary risks of the globally prevalent and concerned CCBs under oxidative water treatment scenarios.


Resveratrol improves diabetic cardiomyopathy by preventing asymmetric dimethylarginine-caused peroxisome proliferator-activated receptor-γ coactivator-1α acetylation.
Cardiac protection of resveratrol is related to the improvement of mitochondrial function through sirtuin1 (SIRT1) activation and peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) deacetylation. Asymmetric dimethylarginine (ADMA) as an endogenous inhibitor of nitric oxide synthases is associated with diabetic cardiovascular complications and has a cross-talk with lysine acetylation. This study was to determine whether resveratrol reverses ADMA's pathogenic role in diabetic cardiomyopathy and elucidate the underlying mechanisms in type 2 diabetic (T2DM) rats and cardiomyocytes. T2DM Rats were induced by high-fat diet plus small-dose streptozotocin injection (35 mg/kg). Resveratrol was given by gavage (50 mg/kg/d) to some rats for 16w. Cardiac function was measured by echocardiography, and PGC-1α acetylation was detected by immunoprecipitation. Mitochondrial DNA and ATP contents were analyzed to evaluate mitochondrial biogenesis and function. Endogenous ADMA accumulation and its signal disorders were associated with cardiac and mitochondrial dysfunctions in accompany with increased PGC-1α acetylation and decreased PGC-1α expression in the myocardium of T2DM rats compared with control rats. Resveratrol treatment attenuated ADMA accumulation, cardiac and mitochondrial dysfunctions in parallel with reversing altered PGC-1α expression and acetylation in the myocardium of T2DM rats. Exogenous ADMA not only reproduced mitochondrial dysfunction and cardiac hypertrophy but also reduced PGC-1α expression and enhanced PGC-1α acetylation in accompany of down-regulating SIRT1 and up-regulating acetyltransferase expression, all of which could be prevented by resveratrol pretreatment in cardiomyocytes. These results indicate that ADMA promotes PGC-1α acetylation as a potential therapeutic target for resveratrol of management diabetic cardiomyopathy in T2DM rats.


Pollutant plume delineation from tree core sampling using standardized ranks.
There are currently contradicting results in the literature about the way chloroethene (CE) concentrations from tree core sampling correlate with those from groundwater measurements. This paper addresses this issue by focusing on groundwater and tree core datasets in CE contaminated site, Czech Republic. Preliminary analyses revealed strongly and positively skewed distributions for the tree core dataset, with an intra-tree variability accounting for more than 80% of the total variability, while the spatial analyses based on variograms indicated no obvious spatial pattern for CE concentration. Using rank transformation, it is shown how the results were improved by revealing the initially hidden spatial structure for both variables when they are handled separately. However, bivariate analyses based on cross-covariance functions still failed to indicate a clear spatial correlation between groundwater and tree core measurements. Nonetheless, tree core sampling and analysis proved to be a quick and inexpensive semi-quantitative method and a useful tool.


Cyto-3D-print to attach mitotic cells.
The Cyto-3D-print is an adapter that adds cytospin capability to a standard centrifuge. Like standard cytospinning, Cyto-3D-print increases the surface attachment of mitotic cells while giving a higher degree of adaptability to other slide chambers than available commercial devices. The use of Cyto-3D-print is cost effective, safe, and applicable to many slide designs. It is durable enough for repeated use and made of biodegradable materials for environment-friendly disposal.


Determination of selected oxygenated polycyclic aromatic hydrocarbons (oxy-PAHs) in diesel and air particulate matter standard reference materials (SRMs).
Oxygenated polycyclic aromatic hydrocarbons (oxy-PAHs) have recently received much attention in discussions regarding the negative impacts of particulate matter (PM) on human health and the environment. The National Institute of Standards and Technology provides several environmental matrix standard reference materials (SRMs) with certified and reference values for polycyclic aromatic hydrocarbons (PAHs) and nitrated PAHs. In this study, the concentrations of oxygenated PAHs are determined in three air PM SRMs (1649b, 1648a, and 2786) and three diesel PM SRMs (1650b, 2975, and 1975) using two independent gas chromatography-mass spectrometry methods. Concentrations of oxy-PAHs were at the milligrams per kilogram level with higher overall concentrations in diesel PM (up to 50 mg/kg for 9,10-anthraquinone). One of the highest oxy-PAH concentrations (up to 5 mg/kg) measured in the air particulate SRMs was for 7,12-benz[a]anthracenquinone. These results suggest that oxygenated PAHs should not be neglected in the analysis of PM as their concentrations can be as high as those of some PAHs and are one to two orders of magnitude higher than those for nitro-PAHs.


A 1H-NMR study of bovine casein micelles; influence of pH, temperature and calcium ions on micellar structure.
The influence of pH, temperature and Ca depletion on bovine casein micelle suspensions in D2O containing simulated milk ultrafiltrate was studied by 1H-NMR spectroscopy. In the pH range of 5.8-7.5 the spectrum of the micelles showed very little pH dependence, indicating that no changes occurred in the dynamic behaviour of the proteins constituting the micelle. The NMR spectrum of casein micelles was strongly temperature dependent, particularly in the temperature range of 60-98 degrees C. Increase in temperature resulted in a strong increase in spectral intensity concomitant with changes in the spectral characteristics. In micelle suspensions these changes were reversible, and indicated that at elevated temperatures the rigid structure of the casein micelle started to melt, leading to an increased mobility of appreciable parts of the proteins in the micelle. Ca depletion of the casein micelles by addition of EDTA resulted in an increase in spectral intensity, which arose from the presence of casein components in the serum phase. The spectrum of these serum phase particles resembled closely the spectrum of a solution of total casein in simulated milk ultrafiltrate and was quite different from the spectrum of casein micelles. The implications of these results with respect to models of the structure of bovine casein micelles are discussed.


Phytotoxicity of short-term exposure to excess zinc or copper in Scots pine seedlings in relation to growth, water status, nutrient balance, and antioxidative activity.
The toxic effects of heavy metals pose a significant threat to the productivity and stability of forest ecosystems. Changes in the agrochemical properties of polluted forest soils due to global climate changes can increase the bioavailability of previously immobilized heavy metals. To test this hypothesis, we studied the effects of short-term shock exposure to ZnSO4 (50, 150, 300 μM) or CuSO4 (2.5, 5, 10 μM) in hydroculture on 4- to 6-week-old seedlings of Scots pine (Pinus sylvestris L.) with well-developed root systems. The effects of the excess heavy metals on mineral nutrients and the functioning of low-molecular-weight antioxidants and glutathione in protecting plants from oxidative damage were studied. Even short-term exposure to exogenous metals led to their rapid accumulation in the root system and their subsequent transport to aboveground organs. An increase in the 4-hydroxyalkenals content in seedling needles exposed to excess Cu led to an increase in the content of proanthocyanidins and catechins, which act as scavengers of reactive oxygen species. The impact of both metals led to the rapid development of mineral nutrient imbalances in the seedlings, which were most pronounced in the presence of excess Zn. Exposure to excess Zn led to a disruption in the translocation of Fe and a decrease in the Fe content in the needles. The most dramatic consequence of Zn exposure was the development of Mn deficiency in the roots, which was the likely cause of the inhibition of phenolic compound synthesis. A deficiency in phenolic compounds can have serious environmental consequences for pine populations that are at risk of contamination by Zn and Cu salts.


Percutaneous absorption of neat and aqueous solutions of 2-butoxyethanol in volunteers.
To study the influence of the presence of water on the dermal absorption of 2-butoxyethanol (BE) in volunteers. Six male volunteers were dermally exposed to 50%, 90% or neat w/w BE for 4 h on the volar forearm over an area of 40 cm(2). An inhalation exposure with a known input rate and duration served as a reference dosage. The dermal absorption parameters were calculated from 24-h excretion of total (free + conjugated) butoxyacetic acid (BAA) in urine and BE in blood, measured after both inhalation and dermal exposure. The dermal absorption of BE from aqueous solutions was markedly higher than that of neat BE. The time-weighted average dermal fluxes were calculated from the urine and blood data and expressed in milligrammes per square centimetre per hour. The dermal fluxes obtained from cumulative 24-h excretion of BAA amounted to 1.34+/-0.49, 0.92+/-0.60 and 0.26+/-0.17 mg cm(-2) h(-1 )for 50%, 90% and neat BE, respectively. The dermal fluxes calculated from the BE blood data amounted to 0.92+/-0.34 and 0.74+/-0.25 mg cm(-2) h(-1 )for 50% and 90% BE, respectively. The permeation rates into the blood reached a plateau between 60 and 120 min after the start of exposure, indicating achievement of steady-state permeation. The apparent permeability coefficient K(p), was 1.75+/-0.53x10(-3) and 0.88+/-0.42x10(-3) cm h(-1 )for 50% and 90% BE, respectively. The percutaneous absorption of BE from aqueous solution increased markedly when compared with neat BE. Even water content as low as 10% led to an approximate fourfold increase in the permeation rates. These findings are important for the health risk assessment of occupational exposure to BE, since BE is commonly used in mixtures that contain water. Exposure to aqueous solutions of 50% and 90% of BE may result in substantial skin absorption: if a 60-min skin contact of 1,000 cm(2) is assumed, dermal uptake would be four-times higher than the pulmonary uptake of an 8-h occupational exposure at a TLV of 100 mg m(-3). This clearly justifies the skin notation for BE. For the purpose of biological monitoring, both BE in blood and BAA in urine were shown to be reliable indicators of exposure.


Regulation of phorbol ester-mediated TRAF1 induction in human colon cancer cells through a PKC/RAF/ERK/NF-kappaB-dependent pathway.
Tumor necrosis factor (TNF) receptor-associated factors (TRAFs) are cytoplasmic adapter proteins that link a wide variety of cell surface receptors to the apoptotic signaling cascade. The purpose of this study was to delineate the signaling pathways and TRAF1 promoter elements responsible for phorbol ester-mediated TRAF1 induction in human colon cancers. Here, we found that the PKC activators, phorbol 12-myristate 13-acetate (PMA) and bryostatin I, induced TRAF1 mRNA expression; pretreatment with actinomycin D blocked PMA-mediated TRAF1 expression suggesting induction at the transcriptional level. In contrast, expression of other TRAFs (TRAF2, 3 and 4) was minimally altered by PMA. Various PKC isoform-selective inhibitors blocked PMA-mediated TRAF1 mRNA and promoter stimulation; rottlerin, a selective PKCdelta inhibitor, had no effect suggesting that Ca(2+)-dependent PKC isoforms (e.g., PKCalpha and betaI) play a role in TRAF1 regulation. In addition, the MEK/ERK inhibitors, PD98059 and UO126, suppressed PMA-stimulated TRAF1 promoter activity indicating a role for ERK in TRAF1 induction. Moreover, cotransfection of a dominant-negative Raf-1 (Raf-C4) significantly reduced PMA-stimulated TRAF1 promoter activity whereas transfection of dominant-negative Ras or treatment with Ras inhibitors had minimal to no effect on TRAF1 induction suggesting dependence on Raf, but not Ras, activation. Finally, site-specific mutagenesis of functional NF-kappaB sites (particularly the most proximal site) in the TRAF1 promoter significantly decreased PMA-mediated promoter activity. In conclusion, our results demonstrate selective induction of TRAF1 in human colon cancer cells through a Ca(2+)-dependent PKC/Raf-1/ERK/NF-kappaB-dependent pathway.


Effect of side chain location in (2-aminoethyl)-aminomethyl-2-phenylquinolines as antitumor agents.
Three new derivatives of 2-phenylquinoline having an (2-aminoethyl)aminomethyl group in 7-, 6-, or 4'- (para position of 2-phenyl ring) positions of aromatic system have been prepared. The antitumor activity of these compounds together with 8- or 4- substituted isomers against the HeLa cell is in the order of 8- > 7- > 4- approximately 6- approximately 4'- substituted ones, which is almost in good agreement with that of DNA-binding ability evaluated by means of DNA-titration of UV-VIS spectra, DNA melting experiment, and ethidium displacement assay. Two representative compounds (8- and 4- isomers) are confirmed to have an ability to intercalate into double stranded DNA by topoisomerase I superhelix unwinding assay.


First clinical postmarketing experiences in the treatment of epilepsies with brivaracetam: a retrospective observational multicentre study.
Brivaracetam (BRV) is the latest approved antiepileptic drug and acts as a synaptic vesicle protein 2A ligand. The aim of the present study was to evaluate the efficacy and tolerability of BRV in the clinical setting. Retrospective, observational multicentre study. We retrospectively collected clinical data of patients who received BRV in 10 epilepsy centres using a questionnaire that was answered by the reporting neurologist. Data of 615 epilepsy patients treated with BRV were included in the study. Efficacy regarding seizure frequency and tolerability of BRV were evaluated. Descriptive statistics complemented by X2 contingency tests and effect sizes were performed. Overall, 44% of the patients had a decreased, 38% a stable and 18% an increased seizure frequency. 17% of patients achieved seizure freedom after initiation of BRV. The seizure frequency decreased in 63% of 19 patients with BRV monotherapy. 27% reported adverse effects, but only 10% of patients with monotherapy. Brivaracetam was significantly more often associated with decreased seizure frequency in levetiracetam (LEV) naïve patients (p=0.012), but BRV also led to a decreased seizure frequency in 42% of patients who had been treated with LEV before, including 17% of patients who were completely seizure free. Adverse effects under LEV improved in 62% and deteriorated in 2% of patients after the switch to BRV. At latest follow-up (mean±SD = 26.3±6.5 months), 68% were still on BRV. The present study shows that results of the phase III studies on BRV match data from real life clinical settings. Brivaracetam seems to be a useful alternative in patients who have suffered adverse effects while taking LEV.


Formulation and evaluation of mefenamic acid emulgel for topical delivery.
Emulgels have emerged as a promising drug delivery system for the delivery of hydrophobic drugs. The objective of the study was to prepare emulgel of mefenamic acid, a NSAID, using Carbapol 940 as a gelling agent. Mentha oil and clove oil were used as penetration enhancers. The emulsion was prepared and it was incorporated in gel base. The formulations were evaluated for rheological studies, spreading coefficient studies, bioadhesion strength, skin irritation studies, in vitro release, ex vivo release studies, anti-inflammatory activity and analgesic activity. Formulation F2 and F4 showed comparable analgesic and anti-inflammatory activity when they compared with marketed diclofenac sodium gel. So, it can be concluded that topical emulgel of mefenamic acid posses an effective anti-inflammatory and analgesic activity.


Effect of patients' functional status on satisfaction with outcomes 12 months after elective spine surgery for lumbar degenerative disease.
Comprehensive assessment of quality of care includes patient-reported outcomes, safety of care delivered, and patient satisfaction. The impact of the patient-reported Oswestry Disability Index (ODI) scores at baseline and 12 months on satisfaction with outcomes following spine surgery is not well documented. This study aimed to determine the impact of patient disability (ODI) scores at baseline and 12 months on satisfaction with outcomes following surgery. Analysis of prospectively collected longitudinal web-based multicenter data. Patients undergoing elective surgery for degenerative lumbar disease were entered into a prospective multicenter registry. Primary outcome measures were ODI, North American Spine Society satisfaction (NASS) questionnaire. Baseline and 12-month ODI scores were recorded. Satisfaction at 12 months after surgery was measured using NASS questionnaire. Multivariable proportional odds logistic regression analysis was conducted to determine the impact of baseline and 12-month ODI on satisfaction with outcomes. Of the total 5,443 patients, 64% (n=3,460) were satisfied at a level where surgery met their expectations (NASS level 1) at 12 months after surgery. After adjusting for all baseline and surgery-specific variables, the 12-month ODI score had the highest impact (Wald χ2=1,555, 86% of the total χ2) on achieving satisfaction with outcomes compared with baseline ODI scores (Wald χ2=93, 5% of the total χ2). The level of satisfaction decreases with increasing 12-month ODI score. Greater change in ODI is required to achieve a better satisfaction level when the patient starts with a higher baseline ODI score. Absolute 12-month ODI following surgery had a significant association on satisfaction with outcomes 12 months after surgery. Patients with higher baseline ODI required a larger change in ODI score to achieve satisfaction. No single measure can be used as a sole yardstick to measure quality of care after spine surgery. Satisfaction may be used in conjunction with baseline and 12-month ODI scores to provide an assessment of the quality of spine surgery provided in a patient centric fashion.


The health education of folk midwives: the Serviço Especial de Saúde Pública and mother-child assistance (1940-1960).
The article addresses an endeavor by Serviço Especial de Saúde Pública (Sesp) to train folk midwives who worked in rural communities and to exercise control over these women's activities. The task was entrusted to the agency's prenatal and child hygiene programs, established between the 1940s and 1960s. The agency believed this training and control initiative would be of major importance in helping ensure the success of its project to establish local sanitary services offering mother-child assistance. The goal of working directly with the folk midwives was not only to force them to employ strict hygiene standards when delivering and caring for newborns but especially to use their influence and prestige within these communities to convince the general population to adopt good health practices.


Hydrodynamics-based delivery of the viral interleukin-10 gene suppresses experimental crescentic glomerulonephritis in Wistar-Kyoto rats.
Gene therapy is expected to revolutionize the treatment of kidney diseases. Viral interleukin (vIL)-10 has a variety of immunomodulatory properties. We examined the applicability of vIL-10 gene transfer to the treatment of rats with crescentic glomerulonephritis, a T helper 1 (Th 1) predominant disease. To produce the disease, Wistar-Kyoto rats were injected with a rabbit polyclonal anti-rat glomerular basement membrane antibody. After 3 h, a large volume of plasmid DNA expressing vIL-10 (pCAGGS-vIL-10) solution was rapidly injected into the tail vein. pCAGGS solution was similarly injected into control rats (pCAGGS rats). We confirmed the presence of vector-derived vIL-10 mainly in the liver and observed high serum vIL-10 levels in pCAGGS-vIL-10-injected rats. Compared with the pCAGGS rats, the pCAGGS-vIL-10 rats showed significant therapeutic effects: reduced frequency of crescent formation, decrease in the number of total cells, macrophages, and CD4+ T cells in the glomeruli, decrease in urine protein, and attenuation of kidney dysfunction. Using quantitative real-time polymerase chain reaction, we also observed that this model was Th1-predominant in the glomeruli and that the ratio of the transcripts of CD4, interferon-gamma, tumor necrosis factor-alpha, and monocyte chemotactic protein-1 to the transcripts of glucose-6-phosphate dehydrogenase in the glomeruli were all significantly lower in the pCAGGS-vIL-10 rats than in the pCAGGS rats. These results demonstrate that pCAGGS-vIL-10 gene transfer by hydrodynamics-based transfection suppresses crescentic glomerulonephritis.


Seasonal variations in the total lipid content and fatty acid composition of cultured and wild Crassostrea madrasensis in Sri Lanka.
The marketplace contains a choice of both cultured and wild oysters, often subtle differences in taste and characteristics are observed between wild and cultured oysters. Therefore, seasonal variations of total lipid and total fatty acid compositions were studied in both, wild and cultured populations of Crassostrea madrasensis in Gangewadiya in Kala-Oya estuary and in cultured populations in the Puttalam lagoon in Sri Lanka over a 13 month period. The fatty acid profile was analyzed using Gas Chromatography. The average lipid percentage was 1.28 ± 0.02% and there was no significant difference between cultured and wild oysters (p > 0.05). A total of 17 fatty acids was detected. Significant monthly variations were seen in all fatty acids, except docosapentaenoic acid (DPA/C22.5(n-3)) and docosahexaenoic acid C226(n-3). Significantly high concentrations (p < 0.05) of omega 3 fatty acids were recorded during October while significantly higher omega 6 concentrations were recorded during April (p > 0.05). Saturated fatty acids had the highest concentration followed by polyunsaturated fatty acids and monounsaturated fatty acids. Palmitic acid was the major saturated fatty acid and docosahexaenoic acid was the major polyunsaturated fatty acid. The ratio of total n-3 PUFA to total n-6 PUFA were 2.6, 2.9 and 3.0 in Gangewadiya wild, Gangewadiya cultured and Kandakuliya cultured oysters, respectively. Though there were no significant differences in omega 3 to omega 6 ratios in cultured and wild oysters in Gangewadiya, a significantly higher ratio was calculated for cultured oysters in Kandakuliya. Hence, a discriminant analysis indicated that in terms of the fatty acid composition, cultured C. madrasensis in Kandakuliya is distinct compared to wild and cultured populations from Gangewadiya. Since, both wild and cultured C. madrasensis shared the same waters in Gangewadiya, the lack of differences can be explained. However, in both sites and under wild and cultured conditions, fatty acid ratio confirms the importance of C. madrasensis as an ideal source of omega 3.


Synthesis of peptide bioconjugates.
Bioconjugates play an important role in several fields of biomolecular and biomedicinal sciences. Protein/polypeptide-based conjugates with covalently attached epitope peptides are considered as potential synthetic vaccine candidates and/or target antigens in affinity-based bioassays. This chapter describes the synthesis of two- and three-component bioconjugates using water-soluble branched chain polymeric polypeptides with multiple amino and/or carboxyl groups as macromolecular partners and oligopeptides as epitopes with small molecular mass. The synthetic procedures outline three major strategies for the incorporation of multiple copies of uniformly oriented peptide epitopes. In the first example, chloroacetylated polypeptide is conjugated with SH-peptide to form a thioether linkage. Second, two independent oligopeptides are introduced into a macromolecule by amide and disulfide bonds, respectively. In the third example, a new procedure is reported for the formation of disulfide bridges by the use of Npys-modified polypeptide and SH-peptide.


The use of hybridoma antibodies to probe the antigenic determinants of thyroglobulin.
Using monoclonal antibodies we have begun to define the epitopes of the murine thyroglobulin molecule that elicit autoimmune responses. Based on the principle of complementation, 18 monoclonal antibodies were classified into 8 groups, defined by their reactions with the same or neighboring determinants. Further distinctions between the monoclonals were drawn from comparisons of their cross-reactions with thyroglobulins of other species and their patterns in isoelectric focusing. One low molecular weight fragment of bovine thyroglobulin, which cross-reacts extensively with thyroglobulins of other species, has been isolated and partially characterized.


What sources of organic carbon drive food webs in billabongs? A study based on stable isotope analysis.
We used the stable isotopes of carbon and nitrogen to examine the food webs of three small flood-plain lakes (billabongs) in south-eastern Australia. With few exceptions, stable carbon isotope analysis could not be used to discriminate among the conspicuous potential sources of fringing, emergent or floating vegetation or benthic detritus. These primary sources showed little spatial or temporal variation in δ13C values, with means ranging from-28.5 to-26.8‰ in spring and-29.1 to-25.4‰ in late summer. Submerged vegetation had similar δ13C values to the above sources in spring but showed greater spatial variation and were less 13C-depleted, considerably so in some species, in late summer. Epiphytes and algae were 13C-depleted in spring compared with the other primary sources but became more 13C-enriched in late summer. Mean δ13C values for primary and secondary consumers were not only far more variable (-37.4 to-22.7‰) but in general were more negative than the potential food sources, particularly in spring. Using the combined information from stable carbon and nitrogen isotope analysis, we could narrow down the list of potential primary sources driving food webs in these billabongs. The freshwater crayfish (Cherax) was one of the few taxa that appeared to obtain its biomass carbon from detrital material. Gastropods and leptocerid caddis larvae on emergent or submerged vegetation obtained a mixture of carbon from epiphytes and macrophytes; in both taxa, epiphytes contributed more to biomass carbon than did the macrophytes. However, other common grazers and collector/gatherers sampled from macrophytes, e.g. baetid mayflies, chironomid larvae and atyid shrimps, were often too 13C-depleted even to have derived their biomass carbon solely from epiphytes. Many other primary consumers, including zooplankton, and mussels (Velesunio), and most of the secondary consumers, including water mites (Hydracarina), phantom midge larvae (Chaoborus) and fish, were also 13C-depleted. The enormous biomass of littoral and fringing vegetation could contribute to metazoan food webs in these billabongs only if an additional highly 13C-depleted source was consumed simultaneously. Methane released from billabong sediments could provide such a 13C-depleted carbon source that is re-introduced into metazoan food webs via the consumption of methanotrophic bacteria. Alternatively, food webs in these water bodies are largely driven by an unknown and inconspicuous 13C-depleted primary producer, such as planktonic Chlorophyta.


Evaluation of virus removal efficiency of coagulation-sedimentation and rapid sand filtration processes in a drinking water treatment plant in Bangkok, Thailand.
In order to properly assess and manage the risk of infection by enteric viruses in tap water, virus removal efficiency should be evaluated quantitatively for individual processes in actual drinking water treatment plants (DWTPs); however, there have been only a few studies due to technical difficulties in quantifying low virus concentration in water samples. In this study, the removal efficiency of indigenous viruses was evaluated for coagulation-sedimentation (CS) and rapid sand filtration (RSF) processes in a DWTP in Bangkok, Thailand by measuring the concentration of viruses before and after treatment processes using real-time polymerase chain reaction (qPCR). Water samples were collected and concentrated from raw source water, after CS, and after RSF, and inhibitory substances in water samples were reduced by use of a hydrophobic resin (DAX-8). Pepper mild mottle virus (PMMoV) and JC polyomavirus (JC PyV) were found to be highly prevalent in raw waters, with concentrations of 10(2.88 ± 0.35) and 10(3.06 ± 0.42) copies/L (geometric mean ± S.D.), respectively. Step-wise removal efficiencies were calculated for individual processes, with some variation observed between wet and dry seasons. During the wet season, PMMoV was removed less by CS and more by RSF on average (0.40 log10 vs 1.26 log10, respectively), while the reverse was true for JC PyV (1.91 log10 vs 0.49 log10, respectively). Both viruses were removed similarly during the dry season, with CS removing the most virus (PMMoV, 1.61 log10 and 0.78 log10; JC PyV, 1.70 log10, and 0.59 log10; CS and RSF, respectively). These differences between seasons were potentially due to variations in raw water quality and the characteristics of the viruses themselves. These results suggest that PMMoV and JC PyV, which are more prevalent in environmental waters than the other enteric viruses evaluated in this study, could be useful in determining viral fate for the risk management of viruses in water treatment processes in actual full-scale DWTPs.


A simple method for in vitro preparation of natural killer cells from cord blood.
Cord Blood (CB) has been considered a promising source of natural killer (NK) cells for cellular immunotherapy. However, it is difficult to expand the large numbers of highly pure NK cells from CB without cell sorting and feeder cells/multiple cytokines. In this study, we try to develop a simple, safe and economical method for ex vivo expansion and purification of NK cells from CB without cell sorting and feeder cells/multiple cytokines. The large numbers (mean: 1.59 × 1010) of highly pure (≥90%) NK cells from CB could be obtained through interleukin-2, group A streptococcus and zoledronate stimulation of mononuclear cells using the 21-day culture approach. When compared to resting NK cells, expanded NK cells were a higher expression of activating receptors CD16, NKG2D, NKp30, NKp44, NKp46 and activating markers CD62L and CD69, while the inhibitory receptors, CD158a and CD158b remained largely unchanged. In addition, these cells showed a higher concentration of IFN-γ, TNF-α and GM-CSF secretion and cytotoxicity to K562 cells and acute myeloid leukemia targets than resting NK cells. We develop a simple, safe and economical method to obtain high yield, purity, and functionality NK cells from CB without cell sorting and feeder cells/multiple cytokines.