Influence of maternal infection on offspring immune response in murine larval toxocariasis.
The impact of Toxocara canis infection on the proportion of CD4+ and CD8+ T splenocytes, the serum concentrations of cytokine IFN-gamma and IL-5, and the production of Toxocara-specific antibodies were studied in two C57BL6/J mouse groups and their offspring. The mice were infected with 1000 T. canis eggs on the day of mating (early infection) and on day 14 of pregnancy (late infection). Early infection resulted in a significant increase of CD4+ T-cell subtype, however, a decline in CD8+ in comparison with late infection, as well as with non-infected control. The IFN-gamma serum concentrations decreased in infected mothers after the birth when compared with non-infected mothers, while in the offspring this cytokine was barely or not detectable. In the mothers of both infected groups, the humoral immune response included both parasite-specific IgM and IgG2 antibodies. While IgG1 levels remained constant throughout the whole experiment in mothers with early infection, late-infected mothers became seropositive only 3 weeks after delivery. IgM was not detectable in any offspring. Pups from early-infected mothers had IgG1 antibodies. Conversely, IgG2 was detectable in pups of both experimental infection groups. A significant difference was observed in the amounts of pups/litter of the infected mothers in comparison with the non-infected ones. Only 56% of females after early infection and 79% of those after late infection had a successful pregnancy. However, all mice of the control group produced a litter. The first T. canis larvae were detected in the muscles of the offspring of both groups on day 5 after the birth. These data show the changes in regulatory and cytotoxic immunity mechanisms of the infected mothers and their offspring and the high level of pregnancy loss as a result of larval toxocariasis.


Effect of Uniaxial Tensile Cyclic Loading Regimes on Matrix Organization and Tenogenic Differentiation of Adipose-Derived Stem Cells Encapsulated within 3D Collagen Scaffolds.
Adipose-derived mesenchymal stem cells have become a popular cell choice for tendon repair strategies due to their relative abundance, ease of isolation, and ability to differentiate into tenocytes. In this study, we investigated the solo effect of different uniaxial tensile strains and loading frequencies on the matrix directionality and tenogenic differentiation of adipose-derived stem cells encapsulated within three-dimensional collagen scaffolds. Samples loaded at 0%, 2%, 4%, and 6% strains and 0.1 Hz and 1 Hz frequencies for 2 hours/day over a 7-day period using a custom-built uniaxial tensile strain bioreactor were characterized in terms of matrix organization, cell viability, and musculoskeletal gene expression profiles. The results displayed that the collagen fibers of the loaded samples exhibited increased matrix directionality with an increase in strain values. Gene expression analyses demonstrated that ASC-encapsulated collagen scaffolds loaded at 2% strain and 0.1 Hz frequency showed significant increases in extracellular matrix genes and tenogenic differentiation markers. Importantly, no cross-differentiation potential to osteogenic, chondrogenic, and myogenic lineages was observed at 2% strain and 0.1 Hz frequency loading condition. Thus, 2% strain and 0.1 Hz frequency were identified as the appropriate mechanical loading regime to induce tenogenic differentiation of adipose-derived stem cells cultured in a three-dimensional environment.


Genetic diagnosis of pancreatic cancer.
Genetic analysis of pancreatic juice is a promising aid for the accurate and early diagnosis of pancreatic cancer. K- ras mutation is frequently observed in pancreatic cancer; however, it is not specific for carcinoma because pancreatic adenoma and pancreatitis also show this mutation. Overexpression of p53 protein is solely detected in pancreatic juice from pancreatic cancer patients, but the positivity rate differs among various reports. Telomerase activity in pancreatic juice was detected in 20 of 24 (83.3%) pancreatic cancer patients and in only 1 of 23 (4.3%) pancreatic adenoma patients, while none of 23 (0%) pancreatitis patients showed evident telomerase activity. The relative value for telomerase activity was significantly higher in pancreatic cancer than in adenoma and pancreatitis. Centrosome abnormalities are very frequently seen in pancreas cancer tissues and the detection of these abnormalities is expected to be a potent new diagnostic tool for the genetic analysis of pancreatic juice. Genetic analysis of pancreatic juice will improve the sensitivity and specificity of pancreatic cancer diagnosis.


Bioelectrical homeostasis as a component of acupuncture mechanism.
Low frequency electrical current and super-high frequency electromagnetic field were applied to acupuncture points of stomach meridian in dogs. The stimulation effect on Bioelectrical potentials of 5 acupuncture points of stomach, spleen, liver, kidney, small intestine meridians and non-acupuncture skin zones was studied in conditions of blocked autonomic ganglia or neuro-muscular junctions of the dog. The influence of ganglioblockading and myorelaxating drugs on Bioelectrical potentials of acupuncture points was also researched. The results are discussed from the neurohumoural and bioelectrical hypotheses points of view. The conclusion that both mechanisms of acupuncture supplement each other is drawn. The principle of bioelectrical homeostasis as a component of acupuncture mechanism is proposed. Bioelectrical homeostasis along with other kinds of homeostasis forms a system of first level homeostats which is united into second level homeostat by the autonomic nervous system.


Eustachian tube function and size of the mastoid air cell system in middle ear surgery.
Myringoplasty results in patients with poor tubal function and small cell systems are often disappointing, to both patient and surgeon. Surgical procedures to improve tubal function have been introduced but they all have the disadvantage of being rather elaborate and difficult to perform. The mastoid air cell system on the other hand does not offer the same problems of access. With the concept of creating an air-filled cavity in connection with the middle ear, mastoidectomy was performed in myringoplasty cases with poor tubal function and small (less than 9 cm2 lateral area) cell systems. Preliminary results show a healing rate of 82% in patients managed according to these principles, against 50% in corresponding cases when myringoplasty alone was done.


[Biochemical risk factor for atherosclerosis in young men. Preliminary report].
Atherosclerotic damage of cardiovascular system, including kidneys, is an increasing problem not only in the modern cardiology but also in nephrology and dialysotherapy. The purpose of our study was to evaluate the frequency of particular biochemical risk factors for atherosclerosis (RFfA) in young men being over-pressed with psychological stress because of decisive character of their professional job. There were 68 men in mean age 33.3 +/- 7.4 years, being employed in the state administration for mean 10.9 +/- 7.5 years. On the basis of the results of environmental questionnaire the studied group was divided into two subgroups: subgroup A "passive" (30 persons) which was physically passive (employed in the administration) and subgroup B "active" (38 persons) whose professional job was connected with physical activity. The following biochemical RFfA in the blood were examined: cholesterol and its fractions, triglycerides, urea acid, glucose, fibrinogen, C-reactive protein, homocysteine and antibodies against Chlamydia pneumoniae. The results were as follows: total cholesterol above 200 mg/dl in 63%, LDL cholesterol above 130 mg/dl in 60% and HDL cholesterol below 50 mg/dl in 31% of men. Among "new" RFfA we found "cardiological" (above 0.2 mg/dl) levels of CRP almost in 25% of the whole group. There were no statistically significant differences with respect to the studied RFfA between "active" and "passive" group. On the basis of these preliminary results we presume, that potentially healthy men over-pressed with psychological stress because of decisive character of their professional job, are characterized--independently on the proclaimed physical activity--by high frequency of risk factors for premature atherosclerosis.


Experimentally induced cam impingement in the sheep hip.
Sheep hips have a natural non-spherical femoral head similar to a cam-type deformity in human beings. By performing an intertrochanteric varus osteotomy, cam-type femoro-acetabular impingement (FAI) during flexion can be created. We tested the hypotheses that macroscopic lesions of the articular cartilage and an increased Mankin score (MS) can be reproduced by an experimentally induced cam-type FAI in this ovine in vivo model. Furthermore, we hypothesized that the MS increases with longer ambulatory periods. Sixteen sheep underwent unilateral intertrochanteric varus osteotomy of the hip with the non-operated hip as a control. Four sheep were sacrificed after 14, 22, 30, and 38-weeks postoperatively. We evaluated macroscopic chondrolabral alterations, and recorded the MS, based on histochemical staining, for each ambulatory period. A significantly higher prevalence of macroscopic chondrolabral lesions was found in the impingement zone of the operated hips. The MS was significantly higher in the acetabular/femoral cartilage of the operated hips. Furthermore, these scores increased as the length of the ambulatory period increased. Cam-type FAI can be induced in an ovine in vivo model. Localized chondrolabral degeneration of the hip, similar to that seen in humans (Tannast et al., Clin Orthop Relat Res 2008; 466: 273-280; Beck et al., J Bone Joint Surg Br 2005; 87: 1012-1018), can be reproduced. This experimental sheep model can be used to study cam-type FAI.


Biodiversity and technological potential of lactic acid bacteria isolated from spontaneously fermented amaranth sourdough.
Spontaneous fermented sourdoughs prepared from amaranth flour were investigated for the presence of autochthonous lactic acid bacteria (LAB) predominating microbiota. The doughs were fermented with daily backslopping on a laboratory scale at 30°C for 10 days. LAB counts ranged from 2·60 to 8·54 log CFU g(-1) with a pH declined from 6·2 to 3·8 throughout fermentation. The combined use of randomly amplified polymorphic DNA (RAPD)-PCR analysis and sequence analysis of 16S rRNA was applied for LAB intraspecies differentiation and taxonomic identification, respectively. Enterococcus, Pediococcus and Lactobacillus species were present in amaranth sourdoughs (AS). After the first refreshment step, Lactobacillus plantarum dominated AS until the end of fermentation. In coincidence, when DGGE analysis was performed, the occurrence of a progressive change in bacterial communities allowed the selection of Lact. plantarum as a dominant species. Moreover, technological, functional and safety characteristics of representative RAPD-biotypes were investigated. Lact. plantarum CRL1898 was selected as a potential candidate for gluten-free amaranth sourdough starter. Nowadays, there is an increasing interest in ancient noncereal gluten-free (GF) crops such as amaranth, due to their reported nutritional and health benefits. However, the use of these grains is still limited to traditional foods and bread making processes that are not yet well standardized. Results on the dynamics of autochthonous lactic acid bacteria (LAB) microbiota during laboratory spontaneous amaranth sourdoughs (AS) fermentation will contribute to overcome challenges for GF-fermented products development. In addition, knowledge about LAB diversity involving Enterococcus, Pediococcus and Lactobacillus species, with Lactobacillus plantarum predominating during AS fermentation, and their technological and functional properties provides the basis for the selection of autochthonous strains as starters cultures for novel gluten-free bakery products with enhanced nutritional, sensory and/or safety quality.


Thai-Lepto-on-admission probability (THAI-LEPTO) score as an early tool for initial diagnosis of leptospirosis: Result from Thai-Lepto AKI study group.
Leptospirosis is one of the most important zoonosis in the tropics. Currently, specific laboratory diagnostic test for leptospirosis such as polymerase chain reaction (PCR) or direct culture cannot be applied at the primary care setting especially in the resource- limited countries. Therefore, clinical presentation and laboratory examination are still the primary diagnostic tools for leptospirosis. To detect clinical factors for predicting leptospirosis in suspected cases, and to create a clinical prediction score (THAI-LEPTO) that is practical and easy to use in general practice while awaiting laboratory results. We performed a prospective multicenter study with a development and a validation cohort of patients presenting with clinical suspicion of leptospirosis as per the WHO clinical criteria. The development cohort was conducted at 11 centers in 8 provinces around Thailand. The validation cohort was conducted at 4 centers in 1 province from the Northeastern part of Thailand. Leptospirosis confirmed cases were defined if any one of the tests were positive: microscopic agglutination test, direct culture, or PCR technique. Multivariable logistic regression was used to identify predictors of leptospirosis. The clinical prediction score was derived from the regression coefficients (original) or from the odds ratio values (simplified). We used receiver operating characteristic (ROC) curve analysis to evaluate the diagnostic ability of our score and to find the optimal cutoff values of the score. We used a validation cohort to evaluate the accuracy of our methods. In the development cohort, we enrolled 221 leptospirosis suspected cases and analyzed 211. Among those, 105 (50%) were leptospirosis confirmed cases. In logistic regression adjusted for age, gender, day of fever, and one clinical factor at a time, leptospirosis group had more hypotension OR = 2.76 (95% CI 1.07-7.10), jaundice OR = 3.40 (95%CI 1.48-8.44), muscle pain OR = 2.12 (95%CI 1.06-4.26), acute kidney injury (AKI) OR = 2.90 (95%CI 1.31-6.15), low hemoglobin OR = 3.48 (95%CI 1.72-7.04), and hypokalemia with hyponatremia OR = 3.56 (95%CI 1.17-10.84) than non-leptospirosis group. The abovementioned factors along with neutrophilia and pulmonary opacity were used in the development of the score. The simplified score with 7 variables was the summation of the odds ratio values as follows: hypotension 3, jaundice 2, muscle pain 2, AKI 1.5, low hemoglobin 3, hypokalemia with hyponatremia 3, and neutrophilia 1. The score showed the highest discriminatory power with area under the curve (AUC) 0.82 (95%CI 0.67-0.97) on fever day 3-4. In the validation cohort we enrolled 96 leptospirosis suspected cases and analyzed 92. Of those, 69 (75%) were leptospirosis confirmed cases. The performance of the simplified score with 7 variables at a cutoff of 4 was AUC 0.78 (95%CI 0.68-0.89); sensitivity 73.5; specificity 73.7; positive predictive value 87.8; negative predictive value 58.3. THAI-LEPTO score is a newly developed diagnostic tool for early presumptive diagnosis of leptospirosis in patients presenting with severe clinical suspicion of the disease. The score can easily be applied at the point of care while awaiting confirmatory laboratory results. Each predictor used has been supported by evidence of clinical and pathophysiological correlation.


Funisitis associated with leptospiral abortion in an equine placenta.
Funisitis, inflammation of the umbilical cord, is well recognized in human placentas. This report describes a case of funisitis associated with leptospiral infection in the placenta of a Thoroughbred foal born prematurely. The umbilical cord had diffuse superficial yellow discoloration along its entire length. Microscopic evaluation showed an exudate of neutrophils admixed with fibrin on the surface. Warthin-Starry staining showed spirochetes in the Wharton's jelly of the umbilical cord. A locally extensive, severe placentitis not involving the star and allantoic cystic hyperplasia were the other lesions observed in the allantochorion. Leptospira funisitis is similar to the funisitis of congenital syphilis in humans, although there are some major microscopic differences. In Leptospira funisitis, lesions were limited to the cord surface, whereas in lesions in human umbilical cords with Treponema pallidum infection, the changes are observed mostly around the vessels and in the Wharton's jelly.


Hepatitis B viral replication markers and hepatic fibrosis in untreated chronic hepatitis B virus infection with and without HIV coinfection in Zambia.
To inform novel therapies, a more nuanced understanding of HIV's impact on hepatitis B virus (HBV) natural history is needed, particularly in high burden countries. In Lusaka, Zambia, we compared prospectively recruited adults (18+ years) with chronic HBV infection, with and without HIV. We excluded those with prior antiviral treatment experience or HBV diagnosis due to clinical suspicion (rather than routine testing). We assessed HBV DNA levels, hepatitis B e antigen (HBeAg), CD4 + (if HIV coinfection), and liver disease (transient elastography, serum alanine aminotransferase). In multivariable analyses, we evaluated the association of HIV overall and by level of CD4 + count on these markers. Among 713 adults analyzed, median age was 33 years, 63% were male, and 433 had HBV/HIV coinfection. Median CD4 + count was 200 cells/μl. HBV DNA was greater than 2000 IU/ml for 311 (51.0%) and 227 (32.5%) were HBeAg-positive. 15.5% had advanced fibrosis or cirrhosis. HIV coinfection was associated with five-fold increased HBV DNA levels [adjusted geometric mean ratio, 5.78; 95% confidence interval (CI), 2.29-14.62] and two times the odds of HBeAg-positivity (adjusted odds ratio, 2.54; 95% CI, 1.59-4.08). These associations were significant only at CD4 + counts 100-350 and <100 cells/μl. HIV was not associated with markers of fibrosis or ALT. HIV's impact on HBV natural history likely depends on the degree and duration of immune suppression. There is strong rationale to monitor HBV DNA in people with HBV/HIV coinfection and immune suppression. A better understanding is needed of mechanisms of increased liver-related mortality in people with HBV/HIV coinfection.


Diabetes Mellitus Treated with Medical Nutritional Therapy and Self Blood Glucose Monitoring: A Randomized Controlled Trial.
To assess the effect of Medical Nutritional Therapy (MNT) combined with self blood glucose monitoring (SBGM) in the clinical outcomes of patients with type 2 diabetes. A randomized controlled trial was conducted on patients with uncontrolled, not-using insulin type 2 diabetes at Her Royal Highness Princess Maha Chakri Sirindhorn Medical Center Sixty patients were recruited and randomized equally into intervention group (MNT with SBGM) and control group (usual care). The primary endpoint was improvement of hemoglobin A₁c (HbA₁c) at 12 and 24 weeks. At 12 and 24 weeks the intervention group had significantly improved their glycemic control in comparison to control group (median decrement of HbA₁c at 12 weeks 0.72% vs. 0.15%; p < 0.001 and at 24 weeks 0.85% vs. 0.20%; p < 0.001). Oral hypoglycemic agents were reduced or discontinued in 7 patients in the intervention group and 1 patient in control group who achieved HbA₁c goal after 24 weeks (p = 0.037). After 24 weeks, body weight was significantly decreased from baseline (2.3 kg, p < 0.001) in the intervention group while only non-significant decrease was observed in control group (0.1 kg, p = 0.632). MNT combined with SBGM is an effective non-pharmacological intervention for patients with uncontrolled type 2 diabetes.


How Changes in Reimbursement Practices Influence the Financial Sustainability of Medicine Policy: Lessons Learned from Slovakia.
Objectives: The aim of this study was to review the impact of new reimbursement requirements for medicines in the Slovak Republic based on legislation that came into force in January 2018. Methods: The new legislation was reviewed. The reimbursement dossiers for medicines and health technology assessments and appraisals, justifications for reimbursement decisions, final reimbursement decisions, and all aspects of the appeal mechanisms have been transparently published on the website of the Slovak Ministry of Health and were used for this analysis. Results: Based on the new legislation, there was no need to submit information about relative effectiveness and cost-effectiveness of medicines with less than 1:50,000 eligible patients prior to reimbursement decisions, and the cost-effectiveness threshold has been increased for all other medicines. The estimated impact of the 2-year budget for the 59 medicines submitted for reimbursement without relative effectiveness and cost-effectiveness analysis was €181,273,698, based on the published submission dossiers. The estimated impact of the 2-year budget for the 45 medicines with evidence of relative effectiveness and cost-effectiveness was €178,566,634. In contrast to the easier market access criteria for new original medicines, the new legislation enforces stricter price erosion criteria for generic and biosimilar medicines. Consequently, the number of generic and biosimilar entries was reduced from 242 in 2017 to 224 in 2018. Conclusions: Although some of the new reimbursement applications were not approved by the Ministry of Health, many new medicines were added to the Slovak pharmaceutical reimbursement list based on "balanced assessment" requirements; hence, the system became financially unsustainable. It was necessary to change the legislation from January 2019.


Influence of growth conditions on Candida albicans adhesion, hydrophobicity and cell wall ultrastructure.
The effect of cultivation in 13 media (10 complex, and three synthetic), as well as altering growth conditions, on Candida albicans adhesion, cell surface hydrophobicity and cell wall ultrastructure was studied. Adhesion of C. albicans to buccal epithelial cells (BECs) was significantly modified by all of the factors tested, particularly growth medium. In general, optimal adhesive activity for C. albicans was observed when the cells were grown in defined media (depending on the carbohydrate used) and/or at 25 degrees C. Moreover, significant differences in adhesion to BECs were noted when C. albicans was grown in the same complex medium from different manufacturers and in different batches of medium from the same manufacturer. Electron microscopy revealed significant differences in surface topography and cell wall ultrastructure of C. albicans grown in different media but none of these differences, including presence or absence of an outer floccular layer, appeared to correlate with the adhesive changes noted, which raises questions regarding the location and nature of the Candida adhesin(s). Likewise, cell surface hydrophobicity could not be correlated with adhesion to BECs but may have influenced yeast coadhesion. The results indicate that Candida adhesion is highly dependent upon the cultivation conditions of the yeast cells tested, and may explain discrepancies in the literature regarding the biochemical nature of the surface component(s) responsible for C. albicans adhesion.


Itch regulation of innate and adaptive immune responses in mice and humans.
The E3 ubiquitin ligase Itch has long been appreciated to be a critical suppressor of inflammation, first identified as a regulator of Th2 differentiation and lung inflammation. Recent studies have revealed novel roles for this protein in mouse and human disease, and it is now clear that Itch also limits the function of other lymphocytes, innate immune cells, and nonhematopoietic cells to regulate immunity. In addition to Th2 cells, Itch also regulates Th17 and regulatory T cells. Itch regulates humoral immunity through direct roles in T follicular helper cells and T follicular regulatory cells, and B cells. Furthermore, Itch limits innate immune responses, such as macrophage cytokine production. Through these cell-intrinsic functions, Itch regulates the interplay between innate and adaptive immune cells, resulting in profound autoinflammation in Itch-deficient mice. Whereas Itch deficiency was previously thought to be an extremely rare occurrence humans, whole exome sequencing of patients with unexplained autoimmune disease has revealed at least two additional cases of Itch deficiency in the last year alone, each caused by distinct mutations within the Itch gene. The recent identification of these patients suggests that Itch mutations may be more common than previously thought, and demonstrates the need to understand how this protein regulates inflammation and autoimmune disease.


Recent Evolution of Coastal Tidal Flats and the Impacts of Intensified Human Activities in the Modern Radial Sand Ridges, East China.
The coastal tidal flats of the modern Radial Sand Ridges (RSRs) are typical silt-muddy tidal flats in Central Jiangsu Province. These tidal flats play a critical role in coastline protection and biodiversity conservation, and against storm surges, but have recently been displaying drastic changes in geomorphic dynamics because of human activities. However, a comprehensive understanding of spatiotemporal changes in tidal flats in RSRs remains lacking. Hence, we employed a novel remote sensing method by obtaining the instantaneous high/low tide line positions from over 112 scenes of Landsat satellite images of the study area from 1975 to 2017, which were used to track the recent evolution of the coastal tidal flats in the modern RSRs over the past four decades. We found that the shoreline of the tidal flats showed an advanced seaward trend, and the waterline of the tidal flat presented a gradual process during different periods. The total tidal flat area in the study area showed an obviously decreasing trend overall, and approximately 992 km2 of the tidal flat was lost. We also found that the coastal tidal flats in the modern RSRs were generally undergoing erosion in the low tidal flats, especially in the Northern Swing and Southern Swing areas, while the high tidal flats showed a slowed accretionary change. Land reclamation was the main factor affecting the reduction in the tidal flat area, as the reclamation area has increased by 1300 km2, with an average of 35.14 km2/year. In addition, the erosion of the tidal flats was associated with a reduced sediment supply. Our findings will provide useful information for local managers and researchers to support future environmental management because increasing demand for land and rising sea levels are expected in the future.


Toxicity of Pekinenin C from Euphorbia Pekinensis Radix on Rat Small Intestinal Crypt Epithelial Cell and Its Apoptotic Mechanism.
Pekinenin C is a casbane diterpenoid separated from the root of the traditional Chinese medicine, Euphorbia pekinensis Rupr., which is used as drug for the treatment of edema, ascites, and hydrothorax. Whereas pekinenin C exhibits severe cytotoxicity, the exact toxicity mechanism is unclear. In this study, the effects of pekinenin C on cell inhibition, cell cycle, and cell apoptosis were examined to explain its toxic mechanism. The proliferation of IEC-6 cells was accessed via MTT colorimetric assay after incubated with different concentrations of pekinenin C. Pekinenin C-treated IEC-6 cells labeled with RNase/PI and Annexin V/PI were analyzed by flow cytometric analyses for evaluation of cell cycle distribution and cell apoptosis, respectively. The apoptosis mechanism of pekinenin C on IEC-6 was investigated through assaying the activities of caspase-3, 8, 9 by enzyme-linked immunosorbent assay (ELISA), protein expression of Bax, Bcl-2, apoptosis-inducing factor (AIF), Apaf-1, Fas-associated death domain (FADD) and type 1-associated death domain (TRADD) by Western-blot, mRNA expression of Fas receptor (FasR), Fas ligand (FasL), tumor necrosis factor receptor (TNFR1) and NF-κB by RT-PCR. The results showed that pekinenin C has exhibited obvious IEC-6 cells toxicity and the IC50 value was 2.1 μg·mL(-1). Typical apoptosis characteristics were observed under a transmission electron microscopy, and it was found that pekinenin C could cause G0/G1 phase arrest in IEC-6 cells in a dose-dependent manner and induce apoptosis of IEC-6 cells. Additionally, pekinenin C could increase the expressions of Bax, AIF, Apaf-1, FasR, FasL, TNFR1 and NF-κB, suppress the expression of Bcl-2, FADD and TRADD, then activate caspase-3, 8, 9 cascades, and at last result in apoptosis. These results demonstrated that pekinenin C effectively promoted cell apoptosis, and induced IEC-6 cells apoptosis through both the mitochondrial and death receptor pathways.


Adenine nucleotide carrier protein dysfunction in human disease.
Adenine nucleotide translocase (ANT) is the prototypical member of the mitochondrial carrier protein family, primarily involved in ADP/ATP exchange across the inner mitochondrial membrane. Several carrier proteins evolutionarily related to ANT, including SLC25A24 and SLC25A25, are believed to promote the exchange of cytosolic ATP-Mg2+ with phosphate in the mitochondrial matrix. They allow a net accumulation of adenine nucleotides inside mitochondria, which is essential for mitochondrial biogenesis and cell growth. In the last two decades, mutations in the heart/muscle isoform 1 of ANT (ANT1) and the ATP-Mg2+ transporters have been found to cause a wide spectrum of human diseases by a recessive or dominant mechanism. Although loss-of-function recessive mutations cause a defect in oxidative phosphorylation and an increase in oxidative stress which drives the pathology, it is unclear how the dominant missense mutations in these proteins cause human diseases. In this review, we focus on how yeast was productively used as a model system for the understanding of these dominant diseases. We also describe the relationship between the structure and function of ANT and how this may relate to various pathologies. Particularly, mutations in Aac2, the yeast homolog of ANT, were recently found to clog the mitochondrial protein import pathway. This leads to mitochondrial precursor overaccumulation stress (mPOS), characterized by the toxic accumulation of unimported mitochondrial proteins in the cytosol. We anticipate that in coming years, yeast will continue to serve as a useful model system for the mechanistic understanding of mitochondrial protein import clogging and related pathologies in humans.


Nanoparticle Targeting with Antibodies in the Central Nervous System.
Treatments for disease in the central nervous system (CNS) are limited because of difficulties in agent penetration through the blood-brain barrier, achieving optimal dosing, and mitigating off-target effects. The prospect of precision medicine in CNS treatment suggests an opportunity for therapeutic nanotechnology, which offers tunability and adaptability to address specific diseases as well as targetability when combined with antibodies (Abs). Here, we review the strategies to attach Abs to nanoparticles (NPs), including conventional approaches of chemisorption and physisorption as well as attempts to combine irreversible Ab immobilization with controlled orientation. We also summarize trends that have been observed through studies of systemically delivered Ab-NP conjugates in animals. Finally, we discuss the future outlook for Ab-NPs to deliver therapeutics into the CNS.


Utility of TMS to understand the neurobiology of speech.
According to a traditional view, speech perception and production are processed largely separately in sensory and motor brain areas. Recent psycholinguistic and neuroimaging studies provide novel evidence that the sensory and motor systems dynamically interact in speech processing, by demonstrating that speech perception and imitation share regional brain activations. However, the exact nature and mechanisms of these sensorimotor interactions are not completely understood yet. Transcranial magnetic stimulation (TMS) has often been used in the cognitive neurosciences, including speech research, as a complementary technique to behavioral and neuroimaging studies. Here we provide an up-to-date review focusing on TMS studies that explored speech perception and imitation. Single-pulse TMS of the primary motor cortex (M1) demonstrated a speech specific and somatotopically specific increase of excitability of the M1 lip area during speech perception (listening to speech or lip reading). A paired-coil TMS approach showed increases in effective connectivity from brain regions that are involved in speech processing to the M1 lip area when listening to speech. TMS in virtual lesion mode applied to speech processing areas modulated performance of phonological recognition and imitation of perceived speech. In summary, TMS is an innovative tool to investigate processing of speech perception and imitation. TMS studies have provided strong evidence that the sensory system is critically involved in mapping sensory input onto motor output and that the motor system plays an important role in speech perception.


Changing pattern of Clostridium difficile associated diarrhoea in a tertiary care hospital: a 5 year retrospective study.
Frequent use of broad spectrum antibiotics in hospitalized patients has increased the incidence of Clostridium difficile diarrhoea in recent years. In our tertiary care hospital in north India, C. difficile was responsible for 15 per cent of cases of nosocomial diarrhoea in 1999. A retrospective study was carried out to determine the frequency of C. difficile associated diarrhoea (CdAD) in our hospital, and to assess the effect of awareness among the hospital personnel and control measures taken to present C. difficile infection following the previous report. A retrospective chart review of all suspected cases of CdAD diagnosed at the hospital from January 2001 to December 2005 was done. Clinical specimens comprised 524 stool samples. All the samples were analyzed for C. difficile using culture and ELISA for toxin A and B. Attempts were made to type isolates using antibiogram, SDS-PAGE, gas liquid chromatography (GLC), PCR for toxin A and B gene fragments and restriction fragment length polymorphism (RFLP). A total of 37 (7.1%) specimens were positive for C. difficile toxin (11.2% in 2001, 9.4% in 2002, 8.6% in 2003, 5% in 2004 and 4% in 2005). The highest number of C. difficile toxin positive cases were from stool samples of patients hospitalized in the haematology/oncology ward (67.5% of all positive cases) followed by gastrointestinal surgery, neurology and nephrology wards. Of the C. difficile toxin positive samples, 15 (41%) were also positive for C. difficile culture. The isolates were grouped in to one, 3 and 5 groups using antibiogram, SDS-PAGE and PCR RFLP respectively. We observed an increase in the number of stool specimens tested for C. difficile infection but a decrease in C. difficile positives. A decrease in the number of C. difficile positive cases were noted during the 5 year study period though number of samples tested was increased. This may be due to stringent surveillance and an improved antibiotic policy followed in the hospital.


Photocross-linking of nucleic acids to associated proteins.
Photocross-linking is a useful technique for the partial definition of the nucleic acid-protein interface of nucleoprotein complexes. It can be accomplished by one or two photon excitations of wild-type nucleoprotein complexes or by one photon excitation of nucleoprotein complexes bearing one or more substitutions with photoreactive chromophores. Chromophores that have been incorporated into nucleic acids for this purpose include aryl azides, 5-azidouracil, 8-azidoadenine, 8-azidoguanine, 4-thiouracil, 5-bromouracil, 5-iodouracil, and 5-iodocytosine. The various techniques and chromophores are described and compared, with attention to the photochemical mechanism.


Stringent doxycycline-dependent control of gene activities using an episomal one-vector system.
Conditional expression systems are of pivotal importance for the dissection of complex biological phenomena. Here, we describe a novel EBV-derived episomally replicating plasmid (pRTS-1) that carries all the elements for conditional expression of a gene of interest via Tet regulation. The vector is characterized by (i) low background activity, (ii) high inducibility in the presence of doxycycline (Dox) and (iii) graded response to increasing concentrations of the inducer. The chicken beta actin promoter and an element of the murine immunoglobin heavy chain intron enhancer drive constitutive expression of a bicistronic expression cassette that encodes the highly Dox-sensitive reverse tetracycline controlled transactivator rtTA2(S)-M2 and a Tet repressor-KRAB fusion protein (tTS(KRAB)) (silencer) placed downstream of an internal ribosomal entry site. The gene of interest is expressed from the bidirectional promoter P(tet)bi-1 that allows simultaneous expression of two genes, of which one may be used as surrogate marker for the expression of the gene of interest. Tight down regulation is achieved through binding of the silencer tTS(KRAB) to P(tet)bi-1 in the absence of Dox. Addition of Dox releases repression and via binding of rtTA2(S)-M2 activates P(tet)bi-1.


The role of extruded disk material in thoracolumbar intervertebral disk disease: a retrospective study in 40 dogs.
The objective of the study was to determine the effect of the dispersed or nondispersed form of the extruded disk material (EDM) on the neurological status and surgical outcomes in Hansen thoracolumbar intervertebral disk disease Type I (IVDD-I). Medical records of 40 dogs with IVDD-I were reviewed, including neurologic status on admission, findings on magnetic resonance imaging (MRI), intraoperative findings, and surgical outcomes. In MRI evaluations, EDM was on the right in 16, on the left in 18, and centrally in 6 cases; in all cases, findings were confirmed by surgery. Extruded disk material was localized and classified as dispersed disk (DD) or nondispersed disk (NDD) according to its dispersion in the epidural space on MRI. Twenty-five dogs had DD and 15 had NDD on both MRI and surgery. There was no significant difference between DD and NDD in preoperative neurological status and surgical outcomes (P > 0.05).


The regulation of hematopoietic growth factor production from cord mononuclear cells and its effect on newborn rat hematopoiesis.
Newborns are characterized by significant developmental immaturities in hematopoiesis and phagocytic immunity. Defects in myelopoiesis and thrombopoiesis predispose newborns to peripheral cytopenias, especially during states of increased demand. However, committed myeloid (CFU-GM) and megakaryocytic (CFU-Meg) progenitor cells in circulating cord blood are increased compared to that of adult bone marrow and/or adult peripheral blood. This increase in hematopoietic committed progenitor cells has allowed the use of cord blood for hematopoietic reconstitution following myeloablative therapy in transplantation. Decreased production of GM-CSF, G-CSF, and IL-3 has been demonstrated in cord-versus-adult mononuclear cells with a similar decrease in their mRNA transcript expression. Other growth factors, however, including steel factor, are increased in the basal state and further increased during stimulation from umbilical-vein endothelial cells versus adult-aortic endothelial cells. The use of single, sequential, or simultaneous administration of both early- and late-acting hematopoietic growth factors, however, has profound effects in the modulation of neonatal-rat hematopoiesis. The combination of G-CSF and early-acting CSFs, such as steel factor or IL-3, induces a significant increase in peripheral neutrophilia and bone marrow progenitor pools.


Spousal loss, children, and the risk of nursing home admission.
OBJECTIVES. Informed by a life course perspective, this study investigates the effects of spousal loss and availability of adult children on elderly husbands' and wives' risk of nursing home entry. Based on longitudinal data from the Health and Retirement Study, we studied 2,116 couples who were community residents in 1998. We estimate proportional hazards models for husbands' and wives' duration to first nursing home admission during 8 years of follow-up. Overall, 438 (20.7%) husbands and 382 (18.1%) wives were institutionalized, and 362 (17.1%) husbands and 701 (33.1%) wives lost their spouse. Accounting for measured covariates, the risk of nursing home entry doubled for men following spousal death, but was unchanged for women. Results indicate that adult children reduced wives' risk of nursing home admission regardless of husbands' vital status, but buffered husbands' risk only after the death of their wives. We uncover suggestive evidence of parent-child gender concordance in children's buffering effect of widowed parents' risk of institutionalization. Discussion. Our findings are consistent with gender variations in spousal caregiving and in husbands' and wives' relative reliance on care from a partner and children. This study provides new evidence on the relationship between institutionalization and family structure among married elderly persons.


Incidence of ANCA-associated vasculitis in a UK mixed ethnicity population.
We aimed to estimate the incidence of ANCA-associated vasculitis in the UK and how this varied by ethnic group. We identified incident cases of ANCA-associated vasculitis between March 2007 and June 2013 in the Nottingham-Derby urban area from medical records using multiple sources. We derived the denominator population from the 2011 census, and we calculated incidence rate ratios using Poisson regression. Overall, we identified 107 cases of ANCA-associated vasculitis, giving an incidence of 23.1 per million person-years (95% CI: 18.9, 27.9). The incidence among the white population was 25.8 per million person-years (95% CI: 21.0, 31.3) and among the black and minority ethnic (BME) population 8.4 per million person-years (95% CI: 3.1, 18.3). After adjustment for age and sex, the difference between ethnic groups was not statistically significant (incidence rate ratio 0.7, 95% CI: 0.3, 1.5, P = 0.3). Overall, the incidence of ANCA-associated vasculitis was similar to other epidemiological studies. Crude incidence rates were lower in the BME than in the white population, but this was partly explained by the older age profile among the white compared with BME population.


Medial Nucleus Accumbens Projections to the Ventral Tegmental Area Control Food Consumption.
Decades of research have shown that the NAc is a critical region influencing addiction, mood, and food consumption through its effects on reinforcement learning, motivation, and hedonic experience. Pharmacological studies have demonstrated that inhibition of the NAc shell induces voracious feeding, leading to the hypothesis that the inhibitory projections that emerge from the NAc normally act to restrict feeding. While much of this work has focused on projections to the lateral hypothalamus, the role of NAc projections to the VTA in the control food intake has been largely unexplored. Using a retrograde viral labeling technique and real-time monitoring of neural activity with fiber photometry, we find that medial NAc shell projections to the VTA (mNAc→VTA) are inhibited during food-seeking and food consumption in male mice. We also demonstrate that this circuit bidirectionally controls feeding: optogenetic activation of NAc projections to the VTA inhibits food-seeking and food intake (in both sexes), while optogenetic inhibition of this circuit potentiates food-seeking behavior. Additionally, we show that activity of the NAc to VTA pathway is necessary for adaptive inhibition of food intake in response to external cues. These data provide new insight into NAc control over feeding in mice, and contribute to an emerging literature elucidating the role of inhibitory midbrain feedback within the mesolimbic circuit.SIGNIFICANCE STATEMENT The medial NAc has long been known to control consummatory behavior, with particular focus on accumbens projections to the lateral hypothalamus. Conversely, NAc projections to the VTA have mainly been studied in the context of drug reward. We show that NAc projections to the VTA bidirectionally control food intake, consistent with a permissive role in feeding. Additionally, we show that this circuit is normally inactivated during consumption and food-seeking. Together, these findings elucidate how mesolimbic circuits control food consumption.


Microwave-assisted synthesis of new benzimidazole derivatives with lipase inhibition activity.
A practical protocol has been used for the synthesis of benzimidazoles. The reaction of iminoester hydrochlorides of phenylacetic with 4,5-dichloro-1,2-phenylenediamine under microwave irradiation leads to the benzimidazole derivatives with good yields and in short reaction times. After the synthesis of benzimidazoles, we synthesized ester and hydrazide derivatives under microwave irradiation with good yields. All compounds were evaluated with regard to pancreatic lipase activity and 3b, 3c, 5a and 6a showed lipase inhibition at various concentrations.


Differential induction of peroxisomal beta-oxidation enzymes by clofibric acid and aspirin in piglet tissues.
Peroxisomal beta-oxidation (POX) of fatty acids is important in lipid catabolism and thermogenesis. To investigate the effects of peroxisome proliferators on peroxisomal and mitochondrial beta-oxidation in piglet tissues, newborn pigs (1-2 days old) were allowed ad libitum access to milk replacer supplemented with 0.5% clofibric acid (CA) or 1% aspirin for 14 days. CA increased ratios of liver weight to body weight (P < 0.07), kidney weight to body weight (P < 0.05), and heart weight to body weight (P < 0.001). Aspirin decreased daily food intake and final body weight but increased the ratio of heart weight to body weight (P < 0.01). In liver, activities of POX, fatty acyl-CoA oxidase (FAO), total carnitine palmitoyltransferase (CPT), and catalase were 2.7-, 2.2-, 1.5-fold, and 33% greater, respectively, for pigs given CA than for control pigs. In heart, these variables were 2.2-, 4.1-, 1.9-, and 1.8-fold greater, respectively, for pigs given CA than for control pigs. CA did not change these variables in either kidney or muscle, except that CPT activity was increased approximately 110% (P < 0.01) in kidney. Aspirin increased only hepatic FAO and CPT activities. Northern blot analysis revealed that CA increased the abundance of catalase mRNA in heart by approximately 2.2-fold. We conclude that 1) POX and CPT in newborn pigs can be induced by peroxisomal proliferators with tissue specificity and 2) the relatively smaller induction of POX in piglets (compared with that in young or adult rodents) may be related to either age or species differences.


The effectiveness of physical and organisational ergonomic interventions on low back pain and neck pain: a systematic review.
Ergonomic interventions (physical and organisational) are used to prevent or reduce low back pain (LBP) and neck pain among workers. We conducted a systematic review of randomised controlled trials (RCTs) on the effectiveness of ergonomic interventions. A total of 10 RCTs met the inclusion criteria. There was low to moderate quality evidence that physical and organisational ergonomic interventions were not more effective than no ergonomic intervention on short and long term LBP and neck pain incidence/prevalence, and short and long term LBP intensity. There was low quality evidence that a physical ergonomic intervention was significantly more effective for reducing neck pain intensity in the short term (ie, curved or flat seat pan chair) and the long term (ie, arm board) than no ergonomic intervention. The limited number of RCTs included make it difficult to answer our broad research question and the results should be interpreted with care. This review, however, provides a solid overview of the high quality epidemiological evidence on the (usually lack of) effectiveness of ergonomic interventions on LBP and neck pain.


Acardiac twin: a systematic review of minimally invasive treatment modalities.
This review of the literature aimed to determine pregnancy outcomes after minimally invasive treatment for occluding vascular supply to acardiac twins. A MEDLINE search was performed until 2002 using the terms 'acardiac', 'acardius', 'twin reversed arterial perfusion (TRAP) sequence' and 'chorangiopagus parasiticus' and 207 articles published in the English-language literature were identified. These articles and the corresponding cited references in English were reviewed. We identified 32 reports involving 74 cases of acardiac twin treated by minimally invasive techniques. Seventy one cases were included for analysis including 40 treated by cord occlusion and 31 by intrafetal ablation. Cord occlusion was first attempted by embolization (n = 5), cord ligation (n = 15), laser coagulation (n = 10), bipolar diathermy (n = 7) and monopolar diathermy (n = 3). Intrafetal ablation was performed by alcohol (n = 5), monopolar diathermy (n = 9), interstitial laser (n = 4) and radiofrequency (n = 13). The overall median gestational ages at treatment and delivery were 21 (range, 14-28) weeks and 36 (range, 19-42) weeks, respectively, with a median treatment-delivery interval of 13 (range, 0-25) weeks. The overall pump twin survival rate was 76% (n = 54). Intrafetal ablation was associated with later median gestational age at delivery (37 vs. 32 weeks, P = 0.04) and higher median treatment-delivery interval (16 vs. 9.5 weeks, P = 0.02) compared with cord occlusion techniques. It was also associated with a lower technical failure rate (13% vs. 35%, P = 0.03), lower rate of premature delivery or rupture of membranes before 32 weeks (23% vs. 58%, P = 0.003) and higher rate of clinical success (77% vs. 50%, P = 0.02) than cord occlusion techniques. There were no statistically significant differences in terms of outcome between the subgroups of fetoscopic- or ultrasound-guided cord occlusion techniques. This review suggests that intrafetal ablation is the treatment of choice for acardiac twins because it is simpler, safer and more effective when compared with the cord occlusion techniques.


Lack of HER-2 gene amplification and association with pathological and clinical characteristics of differentiated thyroid cancer.
Human epidermal growth factor receptor 2 (HER-2) is a well recognized prognostic and predictive factor in breast cancer. However, the role of HER-2 in thyroid cancer remains controversial. The aim of this study was to evaluate HER-2 expression in differentiated thyroid cancer (DTC) and determine whether there is an association with other clinical and pathological characteristics. A total of 69 patients with DTC were investigated, 58 of whom had papillary and 11 follicular carcinomas. HER-2 was detected by immunohistochemical examination on sections from formalin-fixed, paraffin-embedded tumor tissues. Tumors with HER-2 expression classed as +1 and +2 were retested with chromogenic in situ hybridization. Clinicopathological data were retrieved from the hospital records of the patients. HER-2 overexpression was found in 4 (6.9%) of the 58 patients with papillary carcinoma, whereas there was no HER-2 overexpression in any of the 11 cases of follicular carcinoma. There was no association of HER-2 expression with tumor size, pathological grade and cervical lymph node metastasis. In conclusion, there were no HER-2 positive cases of follicular carcinoma and the incidence of HER-2 overexpression in papillary carcinoma was very low. Thus, HER-2 cannot be used routinely as a prognostic or predictive factor in DTC. The expression of other epidermal growth factor receptors in DTC merits further investigation.


The Role of Within-Host Competition for Coexistence in Multiparasitoid-Host Systems.
Multiparasitism (females of multiple species parasitizing the same host) is a ubiquitous phenomenon in parasitoids, yet the role of within-host competition has been mostly ignored in multiparasitoid-host models. Here we study the effect of varying the degree of competition at different life stages: competition over oviposition sites (between-adult competition) and larval competition over resources within the host (within-host competition). We adapt a Nicholson-Bailey model to allow for varying levels of between-adult competition (varying the overlap in species distributions) and within-host competition (varying the number of offspring that can successfully emerge from a host). Surprisingly, while stronger between-adult competition reduces coexistence, stronger within-host competition promotes it. Asymmetric between-adult competition (a fecundity difference between the two species) reduces coexistence when compared to symmetric competition; this can be counteracted by asymmetric within-host competition (within-host competitive advantage of the lower-fecundity species), but only when within-host competition is strong and the correlation between the parasitoids' distributions is intermediate. We discuss our results in the context of the interaction between two parasitoid species, Nasonia vitripennis and Nasonia giraulti, which have strongly correlated distributions and high levels of multiparasitism in the field. We conclude that either low or asymmetric within-host competition is unlikely to explain their coexistence.


Mechanical challenges and cytoskeletal impairments in focal segmental glomerulosclerosis.
Focal segmental glomerulosclerosis (FSGS) is a histologically defined form of kidney injury typically mediated by podocyte dysfunction. Podocytes rely on their intricate actin-based cytoskeleton to maintain the glomerular filtration barrier in the face of mechanical challenges resulting from pulsatile blood flow and filtration of this blood flow. This review summarizes the mechanical challenges faced by podocytes in the form of stretch and shear stress, both of which may play a role in the progression of podocyte dysfunction and detachment. It also reviews how podocytes respond to these mechanical challenges in dynamic fashion through rearranging their cytoskeleton, triggering various biochemical pathways, and, in some disease states, altering their morphology in the form of foot process effacement. Furthermore, this review highlights the growing body of evidence identifying several mutations of important cytoskeleton proteins as causes of FSGS. Lastly, it synthesizes the above evidence to show that a better understanding of how these mutations leave podocytes vulnerable to the mechanical challenges they face is essential to better understanding the mechanisms by which they lead to disease. The review concludes with future research directions to fill this gap and some novel techniques with which to pursue these directions.


Long-acting insulin allergy in a diabetic child.
Insulin allergy has been uncommon since the introduction of human recombinant insulin preparations; the prevalence is 2.4%. Insulin injection could elicit immediate reactions, which are usually induced by an IgE-mediated mechanism, within the first hour after drug administration. In the present study, we describe the case of a child who experienced immediate urticaria after long-acting insulin injection. A 9-year-old girl affected by type I diabetes mellitus referred a history of three episodes of urticaria 30 min after insulin subcutaneous injection. During the first week of insulin therapy, she developed generalized immediate urticaria twice after long-acting insulin glargine first and then once after insulin degludec administration. Symptoms resolved within a few hours after treatment with oral antihistamine. She tolerated rapid insulin lispro. Her personal allergological history was negative. Skin prick tests with degludec, glargine and detemir were performed, showing negative results. Intradermal 1:100000-diluted tests were immediately positive for both degludec and glargine but not for detemir. In light of these findings, detemir was administered without any reaction. Our results show that detemir is tolerated by patients with clinical hypersensitivity reactions to degludec and glargine. Although reactions could be attributable to additives allergy, such as zinc or metacresol, this was excluded since all three preparations contain the same components. So, insulin itself acted as offending allergen. Detemir differs from degludec and glargine in a few aminoacids. Therefore, it is possible that the conformational rather than the linear epitope may be responsible for the reaction. This result suggests integrating intradermal tests in the diagnostic flowchart for insulin allergy. Insulin allergy should always be suspected in patients with immediate symptoms after drug injection. As allergologic work-up, prick by prick test and intradermal test to insulin preparations should be performed. In case of negative results of cutaneous tests, insulin analogs may be administered.


Aging is not a risk factor for femoral and tibial fractures in motor vehicle crashes.
To determine the effect of aging on the occurrence of femoral and tibial fractures during vehicular crashes. The Crash Injury Research and Engineering Network (CIREN), which includes occupants of a vehicle < 8 years old with at least one AIS > or = 3 or two AIS > or = 2 injuries in different body regions, comprised the study population. The occurrence of femoral and tibial fractures during vehicular crashes was analyzed in relation to age and other confounders [gender, BMI, stature, change in velocity (Deltav), restraint use, occupant position (driver vs. passenger) and principal direction of force (PDOF)] using chi2, Mantel-Haenszel chi2 and student t test. Multiple logistic regression (MLR) models were built for the prediction of femoral and tibial fractures with age as the independent variable and possible confounders as co-variates. An alpha = 0.05 was used for all statistics. The incidence of femoral and tibial fractures in the study population (N=1,418) was 23% and 27%, respectively. Univariate analyses revealed a negative association between increasing age and femoral fractures and no association between age and tibial fractures. MLR models revealed no clear effect of increasing age on the occurrence of either femoral or tibial fractures. Obesity, frontal PDOF, and high Deltav affected the occurrence of femoral fractures. Tibial fractures were influenced by occupant position (driver), frontal PDOF, high Deltav and shorter stature. Despite the known changes in bone composition and strength with aging, elderly vehicular occupants do not experience higher odds of incurring femoral and tibial fractures during crashes.


Structural characterization of the closed conformation of mouse guanylate kinase.
Guanylate kinase (GMPK) is a nucleoside monophosphate kinase that catalyzes the reversible phosphoryl transfer from ATP to GMP to yield ADP and GDP. In addition to phosphorylating GMP, antiviral prodrugs such as acyclovir, ganciclovir, and carbovir and anticancer prodrugs such as the thiopurines are dependent on GMPK for their activation. Hence, structural information on mammalian GMPK could play a role in the design of improved antiviral and antineoplastic agents. Here we present the structure of the mouse enzyme in an abortive complex with the nucleotides ADP and GMP, refined at 2.1 A resolution with a final crystallographic R factor of 0.19 (R(free) = 0.23). Guanylate kinase is a member of the nucleoside monophosphate (NMP) kinase family, a family of enzymes that despite having a low primary structure identity share a similar fold, which consists of three structurally distinct regions termed the CORE, LID, and NMP-binding regions. Previous studies on the yeast enzyme have shown that these parts move as rigid bodies upon substrate binding. It has been proposed that consecutive binding of substrates leads to "closing" of the active site bringing the NMP-binding and LID regions closer to each other and to the CORE region. Our structure, which is the first of any guanylate kinase with both substrates bound, supports this hypothesis. It also reveals the binding site of ATP and implicates arginines 44, 137, and 148 (in addition to the invariant P-loop lysine) as candidates for catalyzing the chemical step of the phosphoryl transfer.


The Dialogue Continues: The Future of Public Health Nursing.
This editorial reflects upon the current discussion among nursing academics and educators about the division between individual level care and population health and its implications for the subdiscipline of public health nursing, from the perspective of a current practitioner in the field.


The circadian neuropeptide PDF signals preferentially through a specific adenylate cyclase isoform AC3 in M pacemakers of Drosophila.
The neuropeptide Pigment Dispersing Factor (PDF) is essential for normal circadian function in Drosophila. It synchronizes the phases of M pacemakers, while in E pacemakers it decelerates their cycling and supports their amplitude. The PDF receptor (PDF-R) is present in both M and subsets of E cells. Activation of PDF-R stimulates cAMP increases in vitro and in M cells in vivo. The present study asks: What is the identity of downstream signaling components that are associated with PDF receptor in specific circadian pacemaker neurons? Using live imaging of intact fly brains and transgenic RNAi, we show that adenylate cyclase AC3 underlies PDF signaling in M cells. Genetic disruptions of AC3 specifically disrupt PDF responses: they do not affect other Gs-coupled GPCR signaling in M cells, they can be rescued, and they do not represent developmental alterations. Knockdown of the Drosophila AKAP-like scaffolding protein Nervy also reduces PDF responses. Flies with AC3 alterations show behavioral syndromes consistent with known roles of M pacemakers as mediated by PDF. Surprisingly, disruption of AC3 does not alter PDF responses in E cells--the PDF-R(+) LNd. Within M pacemakers, PDF-R couples preferentially to a single AC, but PDF-R association with a different AC(s) is needed to explain PDF signaling in the E pacemakers. Thus critical pathways of circadian synchronization are mediated by highly specific second messenger components. These findings support a hypothesis that PDF signaling components within target cells are sequestered into "circadian signalosomes," whose compositions differ between E and M pacemaker cell types.


Quercetin induces apoptosis and cell cycle arrest in triple-negative breast cancer cells through modulation of Foxo3a activity.
Quercetin, a plant-derived flavonoid found in fruits, vegetables and tea, has been known to possess bioactive properties such as anti-oxidant, anti-inflammatory and anti-cancer. In this study, anti-cancer effect of quercetin and its underlying mechanisms in triple-negative breast cancer cells was investigated. MTT assay showed that quercetin reduced breast cancer cell viability in a time and dose dependent manner. For this, quercetin not only increased cell apoptosis but also inhibited cell cycle progression. Moreover, quercetin increased FasL mRNA expression and p51, p21 and GADD45 signaling activities. We also observed that quercetin induced protein level, transcriptional activity and nuclear translocation of Foxo3a. Knockdown of Foxo3a caused significant reduction in the effect of quercetin on cell apoptosis and cell cycle arrest. In addition, treatment of JNK inhibitor (SP 600125) abolished quercetin-stimulated Foxo3a activity, suggesting JNK as a possible upstream signaling in regulation of Foxo3a activity. Knockdown of Foxo3a and inhibition of JNK activity reduced the signaling activities of p53, p21 and GADD45, triggered by quercetin. Taken together, our study suggests that quercetin induces apoptosis and cell cycle arrest via modification of Foxo3a signaling in triple-negative breast cancer cells.


TATTOO-ASSOCIATED UVEITIS WITH CHOROIDAL GRANULOMA: A RARE PRESENTATION OF SYSTEMIC SARCOIDOSIS.
The authors report a rare case of systemic sarcoidosis manifesting as bilateral granulomatous panuveitis and multiple tattoo granulomas. Case report. A 30-year-old man presented with headaches, ocular pain, photophobia, and blurred vision. He was found to have significant skin inflammation, predominantly involving areas of tattoos. Fundoscopic examination revealed bilateral granulomatous panuveitis with small focal choroidal lesions. Skin biopsy demonstrated granulomatous inflammation of the dermis with tattoo pigment evident in numerous histiocytes. He was diagnosed with systemic sarcoidosis. The authors report another rare case of tattoo-associated uveitis, in which inflammation is limited to tattooed skin and the uvea. This uncommon presentation may prove informative in elucidating the pathogenesis of systemic sarcoidosis.


Vasopressin Expressed in Hypothalamic CRF Neurons Causes Impaired Water Diuresis in Secondary Adrenal Insufficiency.
Patients with secondary adrenal insufficiency can present with impaired free water excretion and hyponatremia, which is due to the enhanced secretion of vasopressin (AVP) despite increased total body water. AVP is produced in magnocellular neurons in the paraventricular nucleus of the hypothalamus (PVH) and supraoptic nucleus and in parvocellular corticotropin-releasing factor (CRF) neurons in the PVH. This study aimed to elucidate whether magnocellular AVP neurons or parvocellular CRF neurons coexpressing AVP are responsible for the pathogenesis of hyponatremia in secondary adrenal insufficiency. The number of CRF neurons expressing copeptin, an AVP gene product, was significantly higher in adrenalectomized AVP-floxed mice (AVPfl/fl) than in sham-operated controls. Adrenalectomized AVPfl/fl mice supplemented with aldosterone showed impaired water diuresis under ad libitum access to water or after acute water loading. They became hyponatremic after acute water loading, and it was revealed under such conditions that aquaporin-2 (AQP2) protein levels were increased in the kidney. Furthermore, translocation of AQP2 to the apical membrane was markedly enhanced in renal collecting duct epithelial cells. Remarkably, all these abnormalities observed in the mouse model for secondary adrenal insufficiency were ameliorated in CRF-AVP-/- mice that lacked AVP in CRF neurons. Our study demonstrates that CRF neurons in the PVH are responsible for the pathogenesis of impaired water excretion in secondary adrenal insufficiency.


Interactions between cladribine (2-chlorodeoxyadenosine) and standard antileukemic drugs in primary cultures of human tumor cells from patients with acute myelocytic leukemia.
The semiautomated fluorometric microculture cytotoxicity assay (FMCA), based on the measurement of fluorescence generated from cellular hydrolysis of fluorescein diacetate (FDA) to fluorescein in microtiter plates, was used for in vitro evaluation of Cladribine (2-chlorodeoxyadenosine, CdA) interactions with five standard antileukemic drugs: amsacrine (Am), etoposide (VP16), daunorubicin (Dnr), cytosine arabinoside (AraC), and mitoxantrone (Mit). Samples from 31 patients with acute myelocytic leukemia (AML) were tested with continuous drug exposure. A large heterogeneity with respect to cell kill was observed for all combinations tested. An additive model provided a significantly better fit of the data compared to the effect of the most active single agent of the combination (Dmax) only for CdA+AraC. When the frequency of additive and synergistic interactions were calculated according to the multiplicative concept for drug interactions, the highest frequencies were observed for CdA+AraC and CdA+Dnr. This interaction pattern was confirmed by isobologram analysis. Cross-resistance analysis revealed high correlations between CdA and AraC whereas the correlations were weaker between CdA and the other drugs. The highest frequency of synergistic interactions was obtained for AraC+CdA, despite their cross-resistance. Of the non-cross-resistant drugs tested, Dnr appears to be the most effective adjunct to CdA in terms of interactions at the cellular level.


Kinetics of the nanohydrogel flattening at a hydrophobic interface.
Hydrophobitized polysaccharides were designed to form the self-assembled nanohydrogels (hydrogel nanoparticles) in the aqueous conditions. For improving their biocompatibilities, they were decorated with the biomembrane-mimetic 2-methacryloyloxyethyl phosphorylcholine (MPC) polymers. The interfacial roles of the decorated membrane-mimetic nanohydrogels were investigated by choosing MPC branched choresteryl-bearing pullulan (CHP). Tapping-mode atomic force microscopy was used to study its adsorption mechanism on the hydrophobic highly oriented pyrolytic graphite (HOPG) surface in aqueous conditions. Dynamic observation at the interfaces revealed two distinctive patterns: the immobilized nanohydrogel particles and the flatten layers. The flattening (unfolding) kinetics with and without MPC branched nanohydrogel revealed that the flattening energy was at approximately 37 k(B)T. The flattening rate of the MPC decorated nanohydrogels was approximately 1.7 times faster than that without MPC decoration, corresponding to minor reduction of the flattening activation energy.


Localization of tones in a room by moving listeners.
It is difficult to localize the source of a tone in a room because standing waves lead to complicated interaural differences that become uninterpretable localization cues. This paper tests the conjecture that localization improves if the listener can move to explore the complicated sound field over space and time. Listener head and torso movements were free and uninstructed. Experiments at low and high frequencies with eight human listeners in a relatively dry room indicated some modest improvement when listeners were allowed to move, especially at high frequencies. The experiments sought to understand listener dynamic localization strategies in detail. Head position and orientation were tracked electronically, and ear-canal signals were recorded throughout the 9 s of each moving localization trial. The availability of complete physical information enabled the testing of two model strategies: (1) relative null strategy, using instantaneous zeros of the listener-related source angle; and (2) inferred source strategy, using a continuum of apparent source locations implied by the listener's instantaneous forward direction and listener-related source angle. The predicted sources were given weights determined by the listener motion. Both models were statistically successful in coping with a great variety of listener motions and temporally evolving cues.


Workshop on human health impacts of halogenated biphenyls and related compounds.
A workshop on the Human Health Impacts of Halogenated Biphenyls and Related Compounds was held to assess the state of current research on these chemicals and to make recommendations for future studies. Participants discussed results from laboratory animal experiments on PCBs, PBBs, dioxins, and dibenzofurans which demonstrate a common mode of toxicological action while also revealing large variations in toxicological potency both within and between these chemical families. These variations demonstrate the importance of congener-specific analyses in future studies of effects of exposure to these compounds. Results from epidemiological studies of environmentally exposed adult and pediatric populations from the U.S., Japan, and Taiwan and occupationally exposed cohorts from around the world were considered. It was concluded that available evidence did not demonstrate serious adverse effects such as cancer, in exposed adult cohorts but did provide indications of possible neurobehavioral effects in children exposed in utero. In addition, workshop participants described newly developed markers of exposure and techniques for assessing endocrinological, immunological, and neurological effects and suggested these be applied to epidemiological studies of the effects of polyhalogenated compounds. Other recommendations included identification of other cohorts and development of a large registry of exposed individuals; performance of detailed studies of reproductive function and outcomes in exposed populations; and follow up of neurobehavioral effects in offspring of exposed women.


[Experimental Influence of Food Waste Fermentation Broth on the Soil Quality in a Loess Hilly Area].
Loess is widely distributed in northwestern China. Due to the arid climate and rainstorm erosion, lack of nutrients and microorganisms, as well as severe salinization limits the ecosystem carrying capacity of loess soil, which has become one of the major causes of regional land desertification. The fermentation broth derived from food waste usually contains substantial organic acids and nutrients such as nitrogen and phosphorus, and it has the advantages of being easily produced industrially and applied as fertilizer. Hence, this broth has the potential to become a soil amendment for loess soils. This work studied the Lanzhou loess, which is a typical soil of the Loess Plateau of China, fertilized with fermentation broth for the evaluation of physicochemical properties and microbial analyses. After the application of the broth amendment, the total nitrogen, available phosphorus and potassium, and organic matter content increased by 363%, 577%, 308%, and 204%, respectively. After planting grass, including Halogeton arachnoideus Moq. and Medicago sativa L., the comprehensive soil fertility level was further improved and the total salt content of the soil was decreased by 2.3 g·kg-1 and 1.2 g·kg-1, respectively. Meanwhile, the fermentation broth promoted the growth of microorganisms, including bacteria and archaea, which increased by 22 times, and fungi by 8.3 times. Therefore, food waste fermentation broth is conducive to further forming plant-microorganisms symbiosis, improving the ecological environment quality of loess soils.


A cost-effectiveness analysis of hypertrophic cardiomyopathy sudden cardiac death risk algorithms for implantable cardioverter defibrillator decision-making.
To conduct a contemporary cost-effectiveness analysis examining the use of implantable cardioverter defibrillators (ICD) for primary prevention in patients with hypertrophic cardiomyopathy (HCM). A discrete-time Markov model was used to determine the cost-effectiveness of different ICD decision-making rules for implantation. Several scenarios were investigated including the reference scenario of implantation rates according to observed real world practice. A 12-year time horizon with an annual cycle length was used. Transition probabilities used in the model were obtained using Bayesian analysis. The study has been reported according to the Consolidated Health Economic Evaluation Reporting Standards (CHEERS) checklist. Using a 5-year SCD risk threshold of 6% was cheaper than current practice and has marginally better total quality adjusted life years (QALYs). This is the most cost-effective of the options considered, with an incremental cost effectiveness ratio of £834 per QALY. Sensitivity analyses highlighted that this decision is largely driven by what health related quality of life (HRQL) is attributed to ICD patients and time horizon. We present a timely new perspective on HCM ICD cost-effectiveness, using methods reflecting real-world practice. While we have shown that a 6% 5-year SCD risk cut-off provides the best cohort stratification to aid ICD decision-making, this will also be influenced by the particular values of costs and HRQL for subgroups or at a local level. The process of explicitly demonstrating the main factors which drive conclusions from such an analysis will help to inform shared decision-making in this complex area for all stakeholders concerned.


[Effect of the isolation method and staining procedure on the morphology of the virion ribonucleoprotein of Sendai virus].
Electron microscopic examinations of morphology of Sendai virus ribonucleoprotein (RNP) isolated from purified virions by two methods and simultaneous staining of the preparations with uranyl acetate and phosphotungstate acid (PTA) were carried out. The staining method was shown not to influence the kind of nucleoproteins which had been isolated from disrupted virions by sucrose density gradient centrifugation. With both strains RNP appeared as strongly helixed filaments. The staining method, however, strongly influenced morphology of RNP isolated from disrupted virions by equilibrium centrifugation in cesium chloride density gradient. After uranyl acetate staining RNP had an appearance of hard filaments whereas in the same preparation stained with PTA partially and completely unwound spirals of nucleocapsid were found alongside with strongly spiralized structures.


Minimally invasive versus traditional chest surgical approach in patients with reduced renal function.
All surgical access approaches to the chest wall cause a different degree of muscle damage and freeing of substances as myogloblin into the bloodstream thus compromising kidney function. The aim of this study was to evaluate the potential kidney damage in relation to entity of muscle lesions caused by the different surgical approaches. The hematic levels of creatine phosphokinase (CPK), myoglobin, lactate dehydrogenase (LDH), creatinine as well as the amount of the diuresis at different intervals of time were taken of 66 patients who underwent a thoracic surgical operation with diverse surgical access approaches. Surgery determines muscle substances to be freed into the bloodstream. Myoglo-blin levels resulted to be correlated to those of CPK (r=0.83; P<0.00005). Although serum levels of myogloblin are not determined as a routine procedure, high levels of CPK must induce to dose myogloblin. The amount of muscle substances freed depend on the width of the surgical access (r=0.7; P<0.00005) and not upon extension (r=0.36; P=0.18) or duration of surgery. (r=0.4; P=0.093). In patients with a reduced renal function or affected by kidney failure a minimally invasive or thoracoscopic approach is indicated whenever possible in order to reduce the amount of myogloblin in the bloodstream.


[Comparison of retrograde amnesia changes within different injury levels of cerebral concussion in rats].
To investigate the retrograde amnesia changes within different injury levels of cerebral concussion in rats. A metallic pendulum striker device of brain injury was deployed to duplicate CC models of different injury levels within Sprague-Dawley (S-D) rats. The investigated animals were divided into two groups according to classification standard, that is, Pure Cerebral Concussion (PCC) group and Complicated Cerebral Concussion (CCC) group. One control group was used, and each group included 8 animals. The retrograde amnesia of each group was assessed by Morris Water Maze (MWM) Test from 3 days preinjury to 7 days postconcussion. Compared with the control group, the retrograde amnesia was detected within 3 days in PCC group, and 5 days in CCC group after injury. At the same time, the two groups both manifested space recognition deficit. The retrograde amnesia existed in both pure cerebral concussion group and complicated cerebral concussion. Furthermore, the lasting time of retrograde amnesia in animals correlates to the injury level of brain concussion.


Substance P in ovarian carcinoid.
A high content of substance P (124 ng/g wet tissue) was demonstrated by radioimmunoassay in lymph node metastases in a patient with bilateral ovarian carcinoid tumours. The plasma substance P level was elevated but urinary 5-hydroxyindoleacetic acid (5-HIAA) was low. It is suggested that, in some carcinoid patients, an elevated plasma substance P level may help diagnosis and follow-up when urinary 5-HIAA is normal.


Metabolic stability and its role in the discovery of new chemical entities.
Determination of metabolic profiles of new chemical entities is a key step in the process of drug discovery, since it influences pharmacokinetic characteristics of therapeutic compounds. One of the main challenges of medicinal chemistry is not only to design compounds demonstrating beneficial activity, but also molecules exhibiting favourable pharmacokinetic parameters. Chemical compounds can be divided into those which are metabolized relatively fast and those which undergo slow biotransformation. Rapid biotransformation reduces exposure to the maternal compound and may lead to the generation of active, non-active or toxic metabolites. In contrast, high metabolic stability may promote interactions between drugs and lead to parent compound toxicity. In the present paper, issues of compound metabolic stability will be discussed, with special emphasis on its significance, in vitro metabolic stability testing, dilemmas regarding in vitro-in vivo extrapolation of the results and some aspects relating to different preclinical species used in in vitro metabolic stability assessment of compounds.


Coordinate production of PGE2 and IL-1 beta in the gingival crevicular fluid of adults with periodontitis: its relationship to alveolar bone loss and disruption by twice daily treatment with ketorolac tromethamine oral rinse.
The inflammatory mediators prostaglandin E2 (PGE2) and interleukin-1 beta (IL-1 beta) play critical roles in the inflammatory process leading to alveolar bone and connective tissue loss in periodontal disease. Data from a previously published 6-month clinical study demonstrated that twice daily use of 0.1% ketorolac tromethamine oral rinse prevented alveolar bone loss in adults with periodontitis. We further analyzed data from this study to examine the relationship between PGE2. IL-1 beta and bone loss. Patient mean PGE2 and IL-1 beta levels in gingival crevicular fluid (M-GCF) measured throughout the course of the study were directly compared to the maximum amount of alveolar bone height loss observed at a single study site in each patient. The maximum amount of bone loss measured was chosen for the analysis since the pattern of bone loss was clearly episodic in nature. A statistically significant correlation (r = 0.73, p = 0.001) exists between M-GCF PGE2 concentration and the maximum amount of bone height lost at individual patient study sites. The correlation between M-GCF IL-1 beta concentration and maximum bone height lost is also statistically significant (r = 0.66, p = 0.005). Over the 6-month duration of the study, both PGE2 and IL-1 beta were coordinately expressed in the placebo treatment group as reflected in the significant correlation between M-GCF concentrations of the 2 mediators (r = 0.81, p < 0.001). Treatment of patients with 0.1% ketorolac tromethamine twice daily for 6 months resulted in reductions of PGE2 in GCF and a negligible correlation between M-GCF PGE2 and M-GCF IL-1 beta (r = 0.42, p = 0.088). This lack of a strong association between the 2 mediators in the ketorolac treatment group provides a direct biochemical readout of the anti-inflammatory efficacy of ketorolac tromethamine oral rinse in patients with periodontitis. Further studies are warranted to determine the full diagnostic potential of M-GCF levels of PGE2 and IL-1 beta for predicting risk of alveolar bone loss in patients with periodontitis and monitoring periodontal therapy effectiveness.


Supplemental oxygen and sleep at altitude.
Windsor, Jeremy S., and George W. Rodway. Supplemental oxygen and sleep at altitude. High Alt. Med. Biol. 7:307-311, 2006.--The purpose of this study was to examine the effect supplemental oxygen has on the respiratory and cardiovascular system of a mountaineer during sleep at high altitude by using a novel ambulatory, multisensor, continuous monitoring device. Supplemental oxygen was administered to a healthy subject via a nasal demand system (0, 16.7, 33.3, or 50 mL/sec per pulse dose delivered over 1 sec) during the first three nights of sleep at 4900 and 5700 m. Increases in pulse dose resulted in a consistent rise in Sa(O(2)) and a fall in minute ventilation (p < 0.05). The 50-mL pulse dose resulted in the greatest changes, with an increase in Sa(O(2)) from 68.5% to 81% (p < 0.05) and a fall in minute ventilation from 13.1 to 10.9 L/min (p < 0.05) being noted. Changes in Sa(O(2)) and minute ventilation also coincided with a fall in apnea/hypopnea index (AHI). At 4900 m the AHI fell from 12.5-52.3 (breathing air) to 0-7.5 (50-mL oxygen pulse), whereas at 5700 m a decrease from 49.1-80.4 to 3.5-10.0 was observed. No changes in respiratory rate or heart rate were identified when different pulse doses were compared (p < 0.05). The multisensor monitoring device proved to be a highly effective system, demonstrating marked improvements in Sa(O(2)), tidal volume, and AHI in our participant when supplemental oxygen was administered via a nasal demand system.


Sports mega-events and changing world order.
Processes of making, sustaining, reforming, and un-making world orders are constants in global politics and development. Understood in the neo-Gramscian tradition pioneered by Robert Cox, ideas, institutions, and material capabilities combine to shape the range of possibilities for more and less stable orders. Sports mega-events (SMEs)-most prominently, the Olympic Games-have played an underappreciated role in this process. This paper examines the ways in which the Olympics manifested and supported the rise of globalized neoliberal hegemony in the early 1980s, the reconfiguration and erosion of this order through the 1990s and 2000s, and efforts to fundamentally revise this order in the new millennium. Particular emphasis is placed on the dual role of SMEs and the Olympics as manifestations of conspicuous consumption and the pursuit of prestige on the one hand, and as focal points for sanctions campaigns and boycotts on the other.


Chromosome diversity and evolution in Liliaceae.
There is an extensive literature on the diversity of karyotypes found in genera within Liliaceae, but there has been no attempt to analyse these data within a robust phylogenetic framework. In part this has been due to a lack of consensus on which genera comprise Liliaceae and the relationships between them. Recently, however, this changed with the proposal for a relatively broad circumscription of Liliaceae comprising 15 genera and an improved understanding of the evolutionary relationships between them. Thus there is now the opportunity to examine patterns and trends in chromosome evolution across the family as a whole. Based on an extensive literature survey, karyo-morphometric features for 217 species belonging to all genera in Liliaceae sensu the APG (Angiosperm Phylogeny Group) were obtained. Included in the data set were basic chromosome number, ploidy, chromosome total haploid length (THL) and 13 different measures of karyotype asymmetry. In addition, genome size estimates for all species studied were inferred from THLs using a power regression model constructed from the data set. Trends in karyotype evolution were analysed by superimposing the karyological data onto a phylogenetic framework for Liliaceae. Combining the large amount of data enabled mean karyotypes to be produced, highlighting marked differences in karyotype structure between the 15 genera. Further differences were noted when various parameters for analysing karyotype asymmetry were assessed. By examining the effects of increasing genome size on karyotype asymmetry, it was shown that in many but not all (e.g. Fritillaria and all of Tulipeae) species, the additional DNA was added preferentially to the long arms of the shorter chromosomes rather than being distributed across the whole karyotype. This unequal pattern of DNA addition is novel, contrasting with the equal and proportional patterns of DNA increase previously reported. Overall, the large-scale analyses of karyotype features within a well-supported phylogenetic framework enabled the most likely patterns of chromosome evolution in Liliaceae to be reconstructed, highlighting diverse modes of karyotype evolution, even within this comparatively small monocot family.


Is the European Patent Office's approach to assessing obviousness of antibody inventions consistent?
Antibodies can be highly effective in the diagnosis and/or treatment of a wide range of diseases, including certain cancers, viral infections and inflammatory diseases. The development of any new therapeutic antibody is very time consuming, requires significant resources and only a small percentage of initial candidates ever make it onto the market. Patent protection therefore plays a key role in allowing companies to obtain a good return on this investment. However, in Europe, the bar for assessing the inventiveness (obviousness) of antibodies is currently set particularly high, so there is a significant risk that new antibodies may be denied patent protection despite having beneficial properties. This article looks at the rationale behind the European Patent Office's (EPO) current approach to assessing the patentability of antibodies, contrasting it with US practice as well as the EPO's assessment of obviousness in the chemical field. It also explores whether a recent decision by an EPO Appeal Board may help to bring about a change in this practice.


The projection and synaptic organisation of NTS afferent connections with presympathetic neurons, GABA and nNOS neurons in the paraventricular nucleus of the hypothalamus.
Elevated sympathetic nerve activity, strongly associated with cardiovascular disease, is partly generated from the presympathetic neurons of the paraventricular nucleus of the hypothalamus (PVN). The PVN-presympathetic neurons regulating cardiac and vasomotor sympathetic activity receive information about cardiovascular status from receptors in the heart and circulation. These receptors signal changes via afferent neurons terminating in the nucleus tractus solitarius (NTS), some of which may result in excitation or inhibition of PVN-presympathetic neurons. Understanding the anatomy and neurochemistry of NTS afferent connections within the PVN could provide important clues to the impairment in homeostasis cardiovascular control associated with disease. Transynaptic labelling has shown the presence of neuronal nitric oxide synthase (nNOS)-containing neurons and GABA interneurons that terminate on presympathetic PVN neurons any of which may be the target for NTS afferents. So far NTS connections to these diverse neuronal pools have not been demonstrated and were investigated in this study. Anterograde (biotin dextran amine - BDA) labelling of the ascending projection from the NTS and retrograde (fluorogold - FG or cholera toxin B subunit - CTB) labelling of PVN presympathetic neurons combined with immunohistochemistry for GABA and nNOS was used to identify the terminal neuronal targets of the ascending projection from the NTS. It was shown that NTS afferent terminals are apposed to either PVN-GABA interneurons or to nitric oxide producing neurons or even directly to presympathetic neurons. Furthermore, there was evidence that some NTS axons were positive for vesicular glutamate transporter 2 (vGLUT2). The data provide an anatomical basis for the different functions of cardiovascular receptors that mediate their actions via the NTS-PVN pathways.


An observation on subsurface defects of ultra high molecular weight polyethylene due to rolling contact.
Ultra high molecular weight polyethylene (UHMWPE) has been used in artificial joints for a few decades, and wear of UHMWPE has been one of the main problems. Though many other materials have been tested over the years, the best clinical results are still achieved with UHMWPE. This makes the study of UHMWPE, especially in relation to artificial joints, very important. Frequently, more severe wear can be observed in artificial knee joints than in artificial hip joints especially when the flaking-like wear occurs. This flaking-like wear can lead to significant destruction of the artificial knee joint. Macroscopically, artificial knee joints have combinational movements of rolling and sliding in order to simulate the motion of the normal knee joint. The components of motion are separated to make study easier. Fatigue tests of UHMWPE under the rolling contact condition were performed in this study. Three ceramic spheres were rolled over the UHMWPE specimen using 37 degrees C distilled water as a lubricant. The UHMWPE specimen was observed by the scanning acoustic tomography, microscopy, and SEM. Some subsurface defects could be observed by SAT even before experiments. Although the apparent wear is not observed on the surface, there was an increase in the number of observable subsurface cracks in the UHMWPE specimen. This shows that cracks occur under the surface after a 10(7) rolling contact loading, which is very close to the cyclic loading and unloading with very little friction compared to the sliding contact.


The clinical need for clustered AChR cell-based assay testing of seronegative MG.
Trial eligibility in myasthenia gravis (MG) remains largely dependent on a positive autoantibody serostatus. This significantly hinders seronegative MG (SNMG) patients from receiving potentially beneficial new treatments. In a subset of SNMG patients, acetylcholine receptor (AChR) autoantibodies are detectable by a clustered AChR cell-based assay (CBA). Of 99 SNMG patients from two academic U.S. centers, 18 (18.2%) tested positive by this assay. Autoantibody positivity was further validated in 17/18 patients. In a complementary experiment, circulating AChR-specific B cells were identified in a CBA-positive SNMG patient. These findings corroborate the clinical need for clustered AChR CBA testing when evaluating SNMG patients.


Changing Trends Towards Herbal Supplements: Need An Insight Into Safety And Herb-Drug Interaction.
Herbs have been used as sustenance and medicine for a very long time, often in conjunction with other prescribed medications. Even though they are thought to be natural and secure, many of these herbs can interact with other medications and cause potentially dangerous adverse effects or decrease the benefits of the medication. The complex and diverse pharmacological functions carried out by the active ingredients in herbs unavoidably alter the pharmacokinetics of chemical drugs when administered in vivo. Drug transporter expression has a direct impact on how medications are absorbed, distributed, metabolized, and excreted in living organisms. Changes in substrate pharmacokinetics can affect the effectiveness and toxicity of a drug when the active ingredients of a herb inhibit or stimulate the expression of transporters. By reviewing published clinical and preclinical studies, this review aims to raise awareness of herb-drug interactions and discuss their evidence-based mechanisms and clinical consequences. More clinical information on herb-drug interactions is required to make choices regarding patient safety as the incidence and severity of herb-drug interactions are rising due to an increase in the use of herbal preparations globally.This review seeks to increase understanding of herb-drug interactions and explore their evidence-based mechanisms and clinical implications by reviewing published clinical and preclinical studies. The incidence and severity of herb-drug interactions are on the rise due to an increase in the use of herbal preparations worldwide, necessitating the need for more clinical data on these interactions in order to make decisions regarding patient safety. Healthcare workers and patients will become more alert to potential interactions as their knowledge of pharmacokinetic herb-drug interactions grows. The study's objective is to raise readers' awareness of possible interactions between herbal supplements and prescription medications who regularly take them.


Tissue-specific extravasation of albumin-bound Evans blue in hypothermic and rewarmed rats.
The effects of hypothermia and rewarming on endothelial integrity were examined in intestines, kidney, heart, gastrocnemius muscle, liver, spleen, and brain by measuring albumin-bound Evans blue loss from the vasculature. Ten groups of twelve rats, normothermic with no pentobarbital, normothermic sampled at 2, 3, or 4 h after pentobarbital, hypothermic to 20, 25, or 30 degrees C, and rewarmed from 20, 25, or 30 degrees C, were cooled in copper coils through which water circulated. Hypothermic rats were cooled to the desired core temperature and maintained there for 1 h; rewarmed rats were cooled to the same core temperatures, maintained there for 1 h, and then rewarmed. Following Evans blue administration, animals were euthanized with methoxyflurane, tissues removed, and Evans blue extracted. Because hypothermia and rewarming significantly decrease blood flow, organ-specific flow rates for hypothermic and rewarmed tissues were used to predict extravasation. Hypothermia decreased extravasation in tissues with continuous endothelium (brain, muscle) and increased it in tissues with discontinuous endothelium (liver, lung, spleen). All tissues exhibited significant (p < 0.05) differences from normothermic controls. These differences are attributed to a combination of anesthesia, flow, and (or) change in endothelial permeability, suggesting that appropriate choice of organ and temperature would facilitate testing pharmacological means of promoting return to normal perfusion.