Decrease in the rate of secondary amyloidosis in Turkish children with FMF: are we doing better?
Familial Mediterranean fever (FMF) is the most common autoinflammatory disease in the world. The most serious complication of FMF is the development of secondary amyloidosis. Besides genetic factors, environment has been implicated in the development of this complication. The main objective of this study is to analyze whether there has been a substantial decrease of secondary amyloidosis in Turkey and possible effective factors. For this purpose, clinical features of the patients diagnosed with secondary amyloidosis between the years 1978 and 1990 were compared with those diagnosed between 2000 and 2009. Severity scores were determined by the use of a scoring system modified for children. Median ages of the group diagnosed between 1978 and 1990 (n = 115; 12.1% among a total of 947 renal biopsies) and diagnosed after 2000 (n = 19; 2% among a total of 974 renal biopsies) were 12 and 13 years, respectively. There were no significant differences between the two patient groups according to gender, age, age of onset, disease duration, and disease severity. There was, however, a clear decrease in the percentage of biopsies with secondary amyloidosis from 12.1% (1978-1990) to 2% (after 2000; p < 0.001). Our results have shown that there has been a significant decrease in the rate of secondary amyloidosis in Turkey. The main reason for this decrease is better medical care with increased awareness and treatment of the disease. However, we suggest that the improvement of infectious milieu may possibly have had a positive effect on the course of this monogenic disease, since inflammatory pathways related to innate immunity are deregulated.


Palmitoleic acid (n-7) attenuates the immunometabolic disturbances caused by a high-fat diet independently of PPARα.
Palmitoleic acid (PMA) has anti-inflammatory and antidiabetic activities. Here we tested whether these effects of PMA on glucose homeostasis and liver inflammation, in mice fed with high-fat diet (HFD), are PPAR-α dependent. C57BL6 wild-type (WT) and PPAR-α-knockout (KO) mice fed with a standard diet (SD) or HFD for 12 weeks were treated after the 10th week with oleic acid (OLA, 300 mg/kg of b.w.) or PMA 300 mg/kg of b.w. Steatosis induced by HFD was associated with liver inflammation only in the KO mice, as shown by the increased hepatic levels of IL1-beta, IL-12, and TNF-α; however, the HFD increased the expression of TLR4 and decreased the expression of IL1-Ra in both genotypes. Treatment with palmitoleate markedly attenuated the insulin resistance induced by the HFD, increased glucose uptake and incorporation into muscle in vitro, reduced the serum levels of AST in WT mice, decreased the hepatic levels of IL1-beta and IL-12 in KO mice, reduced the expression of TLR-4 and increased the expression of IL-1Ra in WT mice, and reduced the phosphorylation of NF ����B (p65) in the livers of KO mice. We conclude that palmitoleate attenuates diet-induced insulin resistance, liver inflammation, and damage through mechanisms that do not depend on PPAR-α.


Spatial distribution of the four invasive plants and their impact on natural communities' dynamics across the arid and semi-arid environments in northwest Pakistan.
Non-native species are globally successful invaders with negative impacts on vegetation communities' social, economic, and ecological values. Hence, the current research was carried out to assess the spatial distribution patterns and vegetative diversity of the four non-native species in severely invaded areas of the semi-arid parts of northern Pakistan. The research was conducted using data from 1065 plots spread across 165 sites. These sites represented habitats throughout Northern Province, such as farm countryside, highlands, and abandoned places in rural and urban areas. The communities were floristically diverse, represented by 107 plant species, and dominated mainly by annual and perennial life forms with herbaceous habits. Similarly, the floristic structure shows significant variation tested by the χ2 test (P< 0.05) for plant status, life forms, life cycle, and habitat base distribution. In addition, the diversity indices show significant variation having the highest diversity in C-III (P. hysterophorus-dominated sites) and lowest in C-IV (S. marianum-dominated sites, i.e., primarily pure communities), indicating non-native species may increase or decrease site diversity. The diversity communities were further supported by higher quantities of soil nutrients, i.e., organic percentage (2.22 ± 0.04). Altitude, soil nutrients, and texture were shown to be the environmental factors most associated with communities that non-native species had invaded. It is recommended that relevant, additional soil and climatic parameters be integrated into species distribution models to improve our understanding of the ecological niches of different species and to make a collective approach for preserving and conserving native plant communities.


Dysglycemia in the critically ill patient: current evidence and future perspectives.
Dysglycemia in critically ill patients (hyperglycemia, hypoglycemia, glycemic variability and time in range) is a biomarker of disease severity and is associated with higher mortality. However, this impact appears to be weakened in patients with previous diabetes mellitus, particularly in those with poor premorbid glycemic control; this phenomenon has been called "diabetes paradox". This phenomenon determines that glycated hemoglobin (HbA1c) values should be considered in choosing glycemic control protocols on admission to an intensive care unit and that patients' target blood glucose ranges should be adjusted according to their HbA1c values. Therefore, HbA1c emerges as a simple tool that allows information that has therapeutic utility and prognostic value to be obtained in the intensive care unit.


Repairing effects of ICAM-1-expressing mesenchymal stem cells in mice with autoimmune thyroiditis.
The aim of the present study was to determine the repairing effects of intercellular adhesion molecule (ICAM)-1-expressing mesenchymal stem cells (MSCs) in mice with autoimmune thyroiditis. Following induction of an experimental autoimmune thyroiditis (EAT) model, mice were randomly divided into the following groups (n=10 each): i) Normal control; and experimental groups that were subject to EAT induction, including ii) EAT model; and iii) primary MSC; iv) C3H10T1/2/MSC; v) C3H10T1/2-MIGR1/MSC; and vi) C3H10T1/2-MIGR1-ICAM-1/MSC, which were all administered the relevant cells. MSCs were administered via the caudal vein. A blood sample was harvested from the angular vein of each animal 28 days post-treatment and ELISA was used to determine the serum total triiodothyronine, total thyroxine (T4), thyroid-stimulating hormone (TSH), anti-thyroid peroxidase (TPOAb), anti-thyroid microsomal (TMAb) and anti-thyroglobulin (TGAb) antibodies. Hematoxylin and eosin staining was performed to evaluate injury of the thyroid gland by determining the size of the follicle, inflammatory infiltration, colloidal substance retention and epithelial injury. Reverse transcription-quantitative polymerase chain reaction was performed to determine the mRNA expression of interleukin (IL)-4, IL-10, IL-17 and interferon (INF)-γ. Western blot analysis was performed to determine the expression of p38 mitogen-activated protein kinase (p38) and extracellular signal-regulated kinase (ERK). To observe cellular migration in vivo, mice were divided into the following groups, (n=10 each), which were subject to EAT induction: i) CM-DiI-labeled primary MSC; ii) CM-DiI-labeled C3H10T1/2/MSC; iii) CM-DiI-labeled C3H10T1/2-MIGR1/MSC; and iv) CM-DiI-labeled C3H10T1/2-ICAM-1/MSC, which were all administered the relevant cells via the caudal vein. C3H10T1/2-ICAM-1/MSCs were able to ameliorate the expression of T4, TSH, TPOAb, TMAb and TGAb in vivo, attenuate thyroid follicle injury and decrease the splenic index in mice. They were also able to ameliorate the mRNA expression of IL-4, IL-10, IL-17 and INF-γ, and the modulation of the P38 and ERK-signaling pathways in the mouse spleen. Furthermore, ICAM-1 overexpression was able to modulate the nesting of MSCs in the thyroid gland and lung. These findings suggest that C3H10T1/2-ICAM-1/MSC may affect the differentiation, proliferation and migration of immunocytes through modulating the p38 and ERK signaling pathways, and that ICAM-1 may modulate the immunoregulatory effects of MSCs by affecting the migration of MSCs in vivo.


Rac function is crucial for cell migration but is not required for spreading and focal adhesion formation.
Cell migration is commonly accompanied by protrusion of membrane ruffles and lamellipodia. In two-dimensional migration, protrusion of these thin sheets of cytoplasm is considered relevant to both exploration of new space and initiation of nascent adhesion to the substratum. Lamellipodium formation can be potently stimulated by Rho GTPases of the Rac subfamily, but also by RhoG or Cdc42. Here we describe viable fibroblast cell lines genetically deficient for Rac1 that lack detectable levels of Rac2 and Rac3. Rac-deficient cells were devoid of apparent lamellipodia, but these structures were restored by expression of either Rac subfamily member, but not by Cdc42 or RhoG. Cells deficient in Rac showed strong reduction in wound closure and random cell migration and a notable loss of sensitivity to a chemotactic gradient. Despite these defects, Rac-deficient cells were able to spread, formed filopodia and established focal adhesions. Spreading in these cells was achieved by the extension of filopodia followed by the advancement of cytoplasmic veils between them. The number and size of focal adhesions as well as their intensity were largely unaffected by genetic removal of Rac1. However, Rac deficiency increased the mobility of different components in focal adhesions, potentially explaining how Rac - although not essential - can contribute to focal adhesion assembly. Together, our data demonstrate that Rac signaling is essential for lamellipodium protrusion and for efficient cell migration, but not for spreading or filopodium formation. Our findings also suggest that Rac GTPases are crucial to the establishment or maintenance of polarity in chemotactic migration.


Neural development of the zebrafish (Danio rerio) pectoral fin.
The innervation and actuation of limbs have been major areas of research in motor control. Here we describe the innervation of the pectoral fins of the larval zebrafish (Danio rerio) and its ontogeny. Imaging and genetic tools available in this species provide opportunities to add new perspectives to the growing body of work on limbs. We used immunocytological and gross histological techniques with confocal microscopy to characterize the pattern of pectoral fin nerves. We retrogradely labeled fin neurons to describe the distributions of the pectoral fin motor pool in the spinal cord. At 5 days postfertilization, four nerves innervate the pectoral fins. We found that the rostral three nerves enter the fin from the dorsal side of the fin base and service the dorsal and middle fin regions. The fourth nerve enters the fin from the ventral fin base and innervates the ventral region. We found no mediolateral spatial segregation between adductor and abductor cell bodies in the spinal cord. During the larval stage pectoral fins have one adductor and one abductor muscle with an endoskeletal disc between them. As the skeleton and muscles expand and differentiate through postlarval development, there are major changes in fin innervation including extensive elaboration to the developing muscles and concentration of innervation to specific nerves and fin regions. The pattern of larval fin innervation recorded is associated with later muscle subdivision, suggesting that fin muscles may be functionally subdivided before they are morphologically subdivided.


A 4-year follow-up of living unrelated donors of kidney allograft in Iran.
Shortage of deceased donor kidneys has resulted in an increased rate of kidney transplantation from living unrelated donors (LURDs). However, there are concerns about short-term and long-term morbidity of the donors. This study reports the clinical and biochemical factors in a follow-up program of Iranian LURDs, one of the largest reported series of kidney donors. Of 7500 individuals who underwent living donor nephrectomies between 2005 and 2008, a total of 1549 participated in this study. They were followed for 18 to 48 months after the kidney donation. The average time for the first study visit was 316.72 days after donation. The mean age of donors was 30.43 ± 6.16 years old. Men consisted 82.5% of the group. Systolic hypertension was detected in 0.2% and diastolic hypertension in 1% of the LURDs; however, anemia prevalence was as high as 47.2%. Hyperuricemia was found in 21.2% of the LURDs, while proteinuria was seen in 13.7%. Glomerular filtration rate was greater than 90 mL/min in 38.2% of the donors, 60 mL/min to 90 mL/min in 54.5%, and less than 60 mL/min in 7.3%. A GFR less than 45 mL/min was seen in 0.1% of the donors. Data suggested that the LURDs in Iran have an appropriate health condition comparable to other donors in other parts of the world. Considering the high prevalence of hyperuricemia in our population and its importance as a risk factor for kidney failure, monitoring serum uric acid in follow-up programs is suggested.


Mesothelioma in Familial Mediterranean Fever With Colchicine Intolerance: A Case Report and Literature Review.
A 65-year-old Italian physician affected by Familial Mediterranean fever (FMF) was hospitalized due to progressive abdominal enlargement, which had begun 6 months before admission. Physical examination revealed ascites and bilateral leg edema. Abdominal CT scan showed ascitic fluid and extensive multiple peritoneal implants; peritoneal CT-guided biopsy revealed an epithelial-type malignant mesothelioma. The patient's past medical history revealed recurrent episodes of abdominal pain and fever from the age of 2. Clinical diagnosis of FMF was suspected at the age of 25, while genetic analysis, performed at the age of 50, confirmed homozygosity for the M694I mutation in the MEFV gene. Treatment with the first line FMF drug colchicine was started and stopped several times because of worsened leukopenia. The patient in fact had a history of asymptomatic leukopenia/lymphopenia from an early age; the intake of colchicine aggravated his pre-existing problem until the definitive suspension of the drug. As for second-line drugs, canakinumab was first prescribed, but due to prescription issues, it was not possible to be administered. When he was given anakinra, there was a worsening of leukopenia leading to septic fever. Systematic literature review indicates that, in most cases, recurrent peritoneal inflammation results in benign peritoneal fibrosis or less commonly in encapsulating peritonitis. There are only a few reported cases of recurrent peritoneal inflammation progressing from FMF to peritoneal mesothelioma (MST). In such cases, intolerance to colchicine or its erratic intake may lead to long-term recurrent inflammation, which usually precedes the development of the tumor, while pre-existing leukopenia, as in our patient, could also be a factor promoting or accelerating the tumor progression. In conclusion, we suggest that in the presence of intolerance or resistance to colchicine, interleukin (IL)-1 inhibition could suppress peritoneal inflammation and prevent MSTs.


Antimalarial screening via large-scale purification of Plasmodium falciparum Ca2+-ATPase 6 and in vitro studies.
The most severe form of human malaria is caused by the parasite Plasmodium falciparum. Despite the current need, there is no effective vaccine and parasites are becoming resistant to most of the antimalarials available. Therefore, there is an urgent need to discover new drugs from targets that have not yet suffered from drug pressure with the aim of overcoming the problem of new emerging resistance. Membrane transporters, such as P. falciparum Ca(2+)-ATPase 6 (PfATP6), the P. falciparum sarcoplasmic/endoplasmic reticulum Ca(2+)-ATPase (SERCA), have been proposed as potentially good antimalarial targets. The present investigation focuses on: (a) the large-scale purification of PfATP6 for maintenance of its enzymatic activity; (b) screening for PfATP6 inhibitors from a compound library; and (c) the selection of the best inhibitors for further tests on P. falciparum growth in vitro. We managed to heterologously express in yeast and purify an active form of PfATP6 as previously described, although in larger amounts. In addition to some classical SERCA inhibitors, a chemical library of 1680 molecules was screened. From these, we selected a pool of the 20 most potent inhibitors of PfATP6, presenting half maximal inhibitory concentration values in the range 1-9 μm. From these, eight were chosen for evaluation of their effect on P. falciparum growth in vitro, and the best compound presented a half maximal inhibitory concentration of ~ 2 μm. We verified the absence of an inhibitory effect of most of the compounds on mammalian SERCA1a, representing a potential advantage in terms of human toxicity. The present study describes a multidisciplinary approach allowing the selection of promising PfATP6-specific inhibitors with good antimalarial activity.


Contrasts between organic participation in apatite biomineralization in brachiopod shell and vertebrate bone identified by nuclear magnetic resonance spectroscopy.
Unusually for invertebrates, linguliform brachiopods employ calcium phosphate mineral in hard tissue formation, in common with the evolutionarily distant vertebrates. Using solid-state nuclear magnetic resonance spectroscopy (SSNMR) and X-ray powder diffraction, we compare the organic constitution, crystallinity and organic matrix-mineral interface of phosphatic brachiopod shells with those of vertebrate bone. In particular, the organic-mineral interfaces crucial for the stability and properties of biomineral were probed with SSNMR rotational echo double resonance (REDOR). Lingula anatina and Discinisca tenuis shell materials yield strikingly dissimilar SSNMR spectra, arguing for quite different organic constitutions. However, their fluoroapatite-like mineral is highly crystalline, unlike the poorly ordered hydroxyapatite of bone. Neither shell material shows (13)C{(31)P} REDOR effects, excluding strong physico-chemical interactions between mineral and organic matrix, unlike bone in which glycosaminoglycans and proteins are composited with mineral at sub-nanometre length scales. Differences between organic matrix of shell material from L. anatina and D. tenuis, and bone reflect evolutionary pressures from contrasting habitats and structural purposes. The absence of organic-mineral intermolecular associations in brachiopod shell argues that biomineralization follows different mechanistic pathways to bone; their details hold clues to the molecular structural evolution of phosphatic biominerals, and may provide insights into novel composite design.


Acoustic Radiation Force Impulse (ARFI) Elastography and Serological Markers in Assessment of Liver Fibrosis and Free Portal Pressure in Patients with Hepatitis B.
BACKGROUND The aim of this study was to investigate the feasibility of using acoustic radiation force impulse (ARFI) elastography, AST-to-platelet ratio index (APRI), and FIB-4 in assessing liver fibrosis and free portal pressure in patients with hepatitis B. MATERIAL AND METHODS We enrolled 126 patients with hepatitis B who underwent liver surgery at the General Surgery Department of the First Affiliated Hospital of Shihezi University Medical School from February 2013 to August 2015. Preoperatively, shear wave velocity (SWV) of the liver was measured with the Siemens S2000 ultrasound system to reflect liver stiffness. Serological markers were collected and fibrosis indices APRI and FIB-4 were calculated. Intraoperatively, liver tissues were harvested and free portal pressure (FPP) was measured. Postoperatively, fibrosis of liver tissues was pathologically staged. RESULTS The results of SWV, APRI, FIB-4, and FPP were all correlated with the degree of liver fibrosis (Spearman correlation coefficients: r=0.777, P<0.001; r=0.526, P<0.001; r=0.471, P<0.001; p<0.000; r=0.675, p<0.000). Receiver operating characteristic curve (ROC) analysis showed that the areas under the curve (AUC) of ARFI, APRI, and FIB-4 in diagnosing liver fibrosis were 0.830, 0.768, and 0.717, respectively, for stage F≥1; 0.861, 0.773, and 0.754, respectively, for stage F≥2; 0.941, 0.793, and 0.779, respectively, for stage F≥3; and 0.945, 0.783, and 0.754, respectively, for stage F=4. SWV, APRI, and FIB-4 were all correlated with FPP (Pearson correlation coefficients: 0.387, P<0.001; 0.446, P<0.001; 0.419, P<0.001). CONCLUSIONS ARFI, APRI, and FIB-4 can assess liver fibrosis in patients with hepatitis B when assessing the portal venous pressure. The difference in diagnostic efficacy between the 3 was not significant.


Capturing Metabolism-Dependent Solvent Dynamics in the Lumen of a Trafficking Lysosome.
The eukaryotic cell compartmentalizes into spatially confined, membrane-enclosed, intracellular structures ( e. g., organelles, endosomes, and vesicles). Here, peculiar physicochemical properties of the local environment occur and participate in the regulation of ongoing molecular processes. In spite of the huge amount of available environmental probes, experiments on subcellular structures are severely challenged by their three-dimensional (3D) movement. This bottleneck is tackled here by focusing an excitation light beam in a periodic orbit around the structure of interest. The recorded signal is used as feedback to localize the structure position at high temporal resolution: microseconds along the orbit, milliseconds between orbits. The lysosome is selected as the intracellular target, together with 6-acetyl-2-dimethylaminonaphthalene (ACDAN) as probe of the physicochemical properties of the intralysosomal environment. Generalized polarization (GP) analysis of ACDAN emission is used to get a quantitative view on intralysosomal solvent dipolar relaxation. Thus, raster image correlation spectroscopy (RICS) analysis reveals that the ACDAN GP signal is fluctuating in the micro-to-millisecond time range during natural organelle 3D trafficking. We show that ACDAN GP fluctuations are characteristic of lysosomes in living cells, are selectively abolished by lysosomal basification, and depend on metabolic energy in the form of ATP. We argue that intralysosomal ACDAN GP fluctuates according to the ongoing organelle metabolism. Indeed, we report alterations in amplitude and timing of GP fluctuations in a cellular model of lysosomal storage disorder (LSD). The strategy proposed provides insight into the elusive local environment of a trafficking lysosome and supports similar molecular investigations at the subcellular level.


European guidelines on perioperative venous thromboembolism prophylaxis: Neurosurgery.
: Although there are numerous publications addressing venous thromboembolism and its prevention in neurosurgery, there are relatively few high-quality studies to guide decisions regarding thromboprophylaxis. In patients undergoing craniotomy, we recommend that if intermittent pneumatic compression (IPC) is used, it should be applied before the surgical procedure or on admission (Grade 1C). In craniotomy patients at particularly high risk for venous thromboembolism, we suggest considering the initiation of mechanical thromboprophylaxis with IPC preoperatively with addition of low molecular weight heparin (LMWH) postoperatively when the risk of bleeding is presumed to be decreased (Grade 2C). In patients with non-traumatic intracranial haemorrhage, we suggest thromboprophylaxis with IPC (Grade 2C). For patients who have had non-traumatic intracranial haemorrhage, we suggest giving consideration to commencement of LMWH or low-dose unfractionated heparin when the risk of bleeding is presumed to be low (Grade 2C). We suggest continuing thromboprophylaxis until full mobilisation of the patient (Grade 2C). For patients undergoing spinal surgery with no additional risk factors, we suggest no active thromboprophylaxis intervention apart from early mobilisation (Grade 2C). For patients undergoing spinal surgery with additional risk factors, we recommend starting mechanical thromboprophylaxis with IPC (Grade 1C), and we suggest the addition of LMWH postoperatively when the risk of bleeding is presumed to be decreased (Grade 2C).


[The model of the rate of radiation-induced mammalian death based on the determination of delayed consequences of different doses of radiation].
Analyzed were model descriptions of the probability of mammalian lethality and a possible life span reduction consequent to acute and chronic exposures to different dose rates. The proposed model of radiation rate of mammalian death links variations of the coefficient of mammalian death due to acute and prolonged radiation exposures with age. Application of the model to relevant experimental data yielded model coefficients for these periods of exposure. The radiation-modified dependence of lethality coefficients on age makes it possible to obtain fairly simple analytical expressions describing the survival probability long after exposure as a function of dose and dose rate, estimating radiation risk at any time point following exposure, and shortening of mean life span associated with exposure to different doses.


Surveillance of bacterial meningitis in children under 2 y of age in Denmark, 1997-2006.
Worldwide the 3 most common pathogens for bacterial meningitis among infants and young children are Haemophilus influenzae, Streptococcus pneumoniae and Neisseria meningitidis. Denmark included in the National Childhood Vaccination Programme vaccination against H. influenzae type B (Hib) in 1993 and invasive pneumococcal disease as of October 2007. Introduction of the conjugated heptavalent pneumococcal vaccine is, as in the post-Hib vaccination era, expected to change the epidemiology of bacterial meningitis in infants and young children. In 1980 it became mandatory to report suspected cases of bacterial meningitis and the surveillance system was further enhanced for laboratory diagnosed cases of N. meningitis in 1992 and S. pneumoniae in 1996 when a reminder procedure to the physician was issued. In this article we review the epidemiology of 418 notified cases of bacterial meningitis in children <2 y of age in Denmark in the pre-pneumococcal vaccination era 1997-2006 and discuss points of awareness for the future surveillance system.


Giant solitary gastric Peutz-Jeghers polyp mimicking a malignant gastric tumor: the largest described in literature.
A solitary Peutz-Jeghers polyp is defined as a unique polyp occurring without associated mucocutaneous pigmentation or a family history of Peutz-Jeghers syndrome. Gastric solitary localization is a rare event, with only eight reported cases to date. We report herein the case of a 43-year old woman who presented with upper gastrointestinal bleeding, severe anemia, weight loss and asthenia. Endoscopy revealed a giant polypoid tumor with signs of neoplastic invasion of the cardia, with pathological aspect suggesting a Peutz-Jeghers hamartomatous polyp. Computed tomography suggested a malignant gastric tumor and a total gastrectomy was performed. The pathological specimen showed a giant 150/70/50 mm polypoid tumor and immunochemistry established the final diagnostic of a Peutz-Jegers type polyp. This is the largest solitary Peutz-Jeghers gastric polyp reported until now, and the second one mimicking a gastric malignancy with lymph node metastasis.


Evolutionary biology redux.
This article offers a novel, enlightened concept for determining the mechanism of evolution. It is based on homeostasis, which distinguishes life from non-life and as such is the universal mechanism for the evolution of all living organisms. This view of evolution is logical, mechanistic, non-scalar, predictive, testable, and falsifiable, and it illuminates the epistemological relationships between physics and biology, ontogeny and phylogeny, development and aging, ultimate and proximate causation, health and disease. In addition to validating Haeckel's biogenetic law and Lamarckian epigenetics, reflecting the enabling value of the cellular approach, this perspective also expresses the evolutionary process at the cell-molecular level, since the mechanism of cell communication itself is universal in biology, in keeping with a Kuhnian paradigm shift. This approach may even elucidate the nature and evolution of consciousness as a manifestation of the cellular continuum from unicellular to multicellular life. We need such a functional genomic mechanism for the process of evolution if we are to make progress in biology and medicine. Like Copernican heliocentrism, a cellular approach to evolution may fundamentally change humankind's perceptions about our place in the universe.


Low intensity focused ultrasound responsive microcapsules for non-ablative ultrafast intracellular release of small molecules.
Focused ultrasound (FU) is in demand for clinical cancer therapy, but the possible thermal injury to the normal peripheral tissues limits the usage of the ablative FU for tumors with a large size; therefore research efforts have been made to minimize the possible side effects induced by the FU treatment. Non-ablative focused ultrasound assisted chemotherapy could open a new avenue for the development of cancer therapy technology. Here, low intensity focused ultrasound (LIFU) for controlled quick intracellular release of small molecules (Mw ≤ 1000 Da) without acute cell damage is demonstrated. The release is achieved by a composite poly(allylamine hydrochloride) (PAH)/poly-(sodium 4-styrenesulfonate) (PSS)/SiO2 microcapsules which are highly sensitive to LIFU and can be effectively broken by weak cavitation effects. Most PAH/PSS/SiO2 capsules in B50 rat neuronal cells can be ruptured and release rhodamine B (Rh-B) into the cytosol within only 30 s of 0.75 W cm-2 LIFU treatment, as demonstrated by the CLSM results. While the same LIFU treatment shows no obvious damage to cells, as proved by the live/dead experiment, showing that 90% of cells remain alive.


Mortality From Ischemic Heart Disease.
Background Ischemic heart disease (IHD) has been considered the top cause of mortality globally. However, countries differ in their rates and there have been changes over time. Methods and Results We analyzed mortality data submitted to the World Health Organization from 2005 to 2015 by individual countries. We explored patterns in relationships with age, sex, and income and calculated age-standardized mortality rates for each country in addition to crude death rates. In 5 illustrative countries which provided detailed data, we analyzed trends of mortality from IHD and 3 noncommunicable diseases (lung cancer, stroke, and chronic lower respiratory tract diseases) and examined the simultaneous trends in important cardiovascular risk factors. Russia, United States, and Ukraine had the largest absolute numbers of deaths among the countries that provided data. Among 5 illustrative countries (United Kingdom, United States, Brazil, Kazakhstan, and Ukraine), IHD was the top cause of death, but mortality from IHD has progressively decreased from 2005 to 2015. Age-standardized IHD mortality rates per 100 000 people per year were much higher in Ukraine (324) and Kazakhstan (97) than in United States (60), Brazil (54), and the United Kingdom (46), with much less difference in other causes of death. All 5 countries showed a progressive decline in IHD mortality, with a decline in smoking and hypertension and in all cases a rise in obesity and type II diabetes mellitus. Conclusions IHD remains the single largest cause of death in countries of all income groups. Rates are different between countries and are falling in most countries, indicating great potential for further gains. On the horizon, future improvements may become curtailed by increasing hypertension in some developing countries and more importantly global growth in obesity.


Sensing and Processing of DNA Interstrand Crosslinks by the Mismatch Repair Pathway.
DNA interstrand crosslinks (ICLs) that are repaired in non-dividing cells must be recognized independently of replication-associated DNA unwinding. Using cell-free extracts from Xenopus eggs that support neither replication nor transcription, we establish that ICLs are recognized and processed by the mismatch repair (MMR) machinery. We find that ICL repair requires MutSα (MSH2-MSH6) and the mismatch recognition FXE motif in MSH6, strongly suggesting that MutSα functions as an ICL sensor. MutSα recruits MutLα and EXO1 to ICL lesions, and the catalytic activity of both these nucleases is essential for ICL repair. As anticipated for a DNA unwinding-independent recognition process, we demonstrate that least distorting ICLs fail to be recognized and repaired by the MMR machinery. This establishes that ICL structure is a critical determinant of repair efficiency outside of DNA replication.


Micronema granuloma in the gingiva of a horse.
A well-circumscribed proliferative mass was protruding from the body of the mandible of a 4-year-old Appaloosa gelding. The mass was uniformly firm and was ulcerated on the dorsal surface. The horse was euthanatized due to extensive involvement of the jaw and difficulty in eating. Microscopically, the mass consisted of numerous discrete microgranulomas separated by bands of abundant fibrous connective tissue containing lymphocytes, plasma cells, and histiocytic macrophages. Numerous rhabditiform nematodes were in the microgranulomas as well as in the surrounding connective tissue. The nematodes were identified as Micronema species.


Optical resolution of various amino acids using a supported liquid membrane encapsulating a surfactant-protease complex.
We have encapsulated a surfactant-protease complex (the main protease used being alpha-chymotrypsin) in an organic phase of a supported liquid membrane (SLM) for the optical resolution of various amino acids. L-Isomers of amino acids were enantioselectively permeated through the SLM. The mechanism of the amino acid permeation through the SLM was considered to be as follows; an L-amino acid was enantioselectively esterified with ethanol by a surfactant-protease complex encapsulated in the SLM, and the resulting L-amino acid ethyl ester dissolved into the organic phase of the SLM and diffused across the SLM. Another surfactant-alpha-chymotrypsin complex in the receiving phase catalyzed ester hydrolysis to produce the initial L-amino acid and ethanol, which are water-soluble. Thus, the L-amino acid was selectively transported to the receiving phase through the SLM on the basis of the molecular recognition of the surfactant-protease complex in the SLM. It was found that the catalytic activity and enantioselectivity of the surfactant-protease complex governed the permeate flux of amino acids and the enantiomeric excess in the membrane separation.


Characterization of lymphokine fibronectin from guinea pig lymphoid cell culture supernatants.
Fibronectins (FN) in guinea pig lymphoid cell culture supernatants have been studied using a panel of polyclonal and monoclonal anti-FN antibodies to clarify their relationship with macrophage agglutination factor (MAggF), an inflammatory lymphokine sharing many properties with this family of high molecular weight glycoproteins. MAggF contained cellular FN epitopes, and was reversibly bound by antibodies specific for cellular FN. Enzyme-linked immunoassay and inhibition of MAggF activity by monoclonal anti-plasma FN antibodies revealed immunoreactive FN in guinea pig lymphoid cell culture supernatants to share three epitopes with plasma FN and to lack a fourth epitope present in plasma FN. Immunoreactive FN in gelatin-affinity purified lymph node cell culture supernatants was polydisperse; MAggF activity (Mr 410 kD) was associated with only 13% of total immunoreactive FN. Although a low molecular weight FN fragment (Mr 67 kD) was associated with MAggF activity in salt-fractionated peritoneal exudate culture supernatants, it was not possible to generate MAggF activity by limited proteolysis of MAggF-inactive, high molecular weight FN in lymph node cell culture supernatants. We conclude that MAggF is a cellular FN containing a number of epitopes in common with plasma FN and suggest it may be a unique species of cellular FN produced by T lymphocytes involved in initiating delayed hypersensitivity reactions.


Insulin action and binding in isolated hepatocytes from fasted, streptozotocin-diabetic, and older, spontaneously obese rats.
Insulin binding and basal and insulin-stimulated uptake of alpha-aminoisobutyric acid were measured in isolated hepatocytes from young control rats as well as from older spontaneously obese, 72h-starved, and nonketotic streptozotocin-diabetic rats. Isolated hepatocytes from older spontaneously obese rats are similar to those from younger smaller rats in size, maximal insulin responsiveness, the dose-response relationship for insulin-stimulated aminoisobutyrate uptake, and the number and affinity of insulin receptors. Hepatocytes from 72h-fasted rats have similar numbers of insulin receptors per cell as cells from young control animals, but are significantly smaller, have an enhanced basal rate of aminoisobutyrate uptake, and are insulin resistant with regard to maximal insulin-stimulated uptake of aminoisobutyrate at 0.1mm-aminoisobutyrate. Because of the decreased maximal response to insulin, the concentration of insulin that elicits a half-maximal response of aminoisobutyrate uptake is decreased. Hepatocytes from diabetic animals, like those from starved rats, have significantly greater basal rates of aminoisobutyrate uptake; whereas the maximal absolute insulin response is the same as control cells, the percentage response is smaller. These cells bind significantly more insulin than do control cells. The increase in insulin binding is reflected in a shift to the left of the dose-response curve for insulin-stimulated uptake of aminoisobutyrate. These studies indicate that there is no insulin resistance with regard to uptake of aminoisobutyrate in hepatocytes from older obese rats. Furthermore, the insulin resistance observed in hepatocytes from starved rats occurs despite an increase in the number of receptors per unit surface area and cannot be explained by alterations in the interaction between insulin and its receptor. The enhanced insulin binding per unit surface area, however, is reflected in the shift to the left of the dose-response curve for insulin. This is also true for hepatocytes from diabetic animals, in which insulin binding per cell is increased.


In vivo MRI analysis of an inflammatory injury in the developing brain.
Cerebral periventricular white matter injury stands as a leading cause of cognitive, behavioral and motor impairment in preterm infants. There is epidemiological and histopathological evidence demonstrating the role of prenatal or neonatal inflammation in brain injury in preterm infants. In order to define the effect of an inflammatory insult in the developing brain on magnetic resonance (MR) imaging, we obtained high resolution conventional and diffusion MR images of the brain of rat pups after an inflammatory injury. Rat pups were subjected on postnatal day 5 (P5) to a stereotaxic injection of lipopolysaccharide in the corpus callosum and then imaged at 11.7 T on days 0, 2 and 4 following the injury. They were subsequently sacrificed for immunohistochemistry. Diffusion tensor imaging (DTI) acquired at high spatial resolution showed an initial reduction of the apparent diffusion coefficient (ADC) in the white matter. This was followed by an increase in ADC value and in T2 relaxation time constant in the white matter, with an associated increase of radial diffusivity of the corpus callosum, and a 10-fold increase in ventricular size. On histology, these MR changes corresponded to widespread astrogliosis, and decreased proportion of the section areas containing cresyl violet positive stain. The increase in radial diffusivity, typically attributed to myelin loss, occurred in this case despite the absence of myelin at this developmental stage.


A comprehensive model for the treatment of nicotine dependence in a medical setting.
We have summarized the basic components of a comprehensive model for the treatment of nicotine addiction in a medical setting. It is a labor-intensive program with emphasis on individual assessment and the development of individual treatment programs adaptable to all levels of nicotine dependence. We anticipate that this model will be increasingly used by medical centers in the future. As more traditional programs successfully treat the less severely nicotine-dependent patients, more severely dependent smokers are left in need of more intensive services. We have already begun to see this trend in our practice, requiring more counselor time for individual follow-up and referral into our group therapy programs. Adjunctive pharmacologic therapy is an exciting and promising area, but best results include associated behavioral intervention. Reimbursement remains a major impediment to patient involvement in many of these programs. This impediment will be overcome when third-party payers begin to cover these services as endorsed by the Surgeon General and the United States Department of Health and Human Services.


Spatial Dynamic Subspaces Encode Sex-Specific Schizophrenia Disruptions in Transient Network Overlap and its Links to Genetic Risk.
Schizophrenia research reveals sex differences in incidence, symptoms, genetic risk factors, and brain function. However, a knowledge gap remains regarding sex-specific schizophrenia alterations in brain function. Schizophrenia is considered a dysconnectivity syndrome, but the dynamic integration and segregation of brain networks are poorly understood. Recent advances in resting-state fMRI allow us to study spatial dynamics, the phenomenon of brain networks spatially evolving over time. Nevertheless, estimating time-resolved networks remains challenging due to low signal-to-noise, limited short-time information, and uncertain network identification. We adapt a reference-informed network estimation technique to capture time-resolved networks and their dynamic spatial integration and segregation for 315 controls and 193 schizophrenia individuals. We focus on time-resolved spatial functional network connectivity (spFNC), an estimate of network spatial coupling, to study sex-specific alterations in schizophrenia and their links to genomic data. Our findings are consistent with the dysconnectivity and neurodevelopment hypotheses and align with the cerebello-thalamo-cortical, triple-network, and frontoparietal dysconnectivity models, helping to unify them. The potential unification offers a new understanding of the underlying mechanisms. Notably, the posterior default mode/salience spFNC exhibits sex-specific schizophrenia alteration during the state with the highest global network integration and correlates with schizophrenia genetic risk. This dysfunction is reflected in regions with weak functional connectivity to corresponding networks. Our method can effectively capture spatially dynamic networks, detect nuanced schizophrenia effects including sex-specific ones, and reveal the intricate relationship of dynamic information to genomic data. The results also underscore the clinical potential of dynamic spatial dependence and weak connectivity.


Neoplastic transformation induced by adeno 12-SV40 hybrid virus in skin fibroblasts from humans genetically predisposed to cancer.
The DNA hybrid adeno 12-SV40 virus induced neoplastic transformation of human skin fibroblasts derived from individuals with adenomatosis of the colon and rectum (ACR), an inherited cancer syndrome. The line established from the transformed ACR fibroblasts became stable and was initially virus productive, but infectious hybrid virus gradually decreased over serial passages. Cultured skin fibroblasts from normal individuals infected with adeno 12-SV40 also underwent morphological transformation and were virus productive. Although these cells had a prolonged life as compared with uninfected controls, they eventually died. All of the transformed cells contained both adeno 12 and SV40 large tumor antigens, formed large cell aggregates, grew in liquid growth medium above an agar base, and formed colonies in soft agar. The altered ACR cells became heteroploid and produced tumors when transplanted subcutaneously into nude mice. The results suggest that ACR individuals have increased susceptibility to neoplastic transformation by oncogenic tumor viruses.


Dynamic changes in T-wave and QTc interval during tilt table testing: Innocent until proven otherwise.
A16-year-old female underwent tilt table testing, which resulted positive for reflex vasodepressive syncope. 12‑lead ECG during syncope showed T-wave inversion in infero-lateral leads, along with QTc interval increase >100 msec compared to baseline. These abnormalities rapidly disappeared in supine position with resumption of consciousness. Complete cardiac evaluation excluded heart disease. T-wave changes and moderate QTc prolongation are relatively common in young (mainly female) patients undergoing tilt table testing and they appear benign in nature. However, in a minority of cases, on the basis of the clinical context and after an accurate ECG analysis, further examinations may be warranted.


Fraud in a population-based study of headache: prevention, detection and correction.
In medicine, research misconduct is historically associated with laboratory or pharmaceutical research, but the vulnerability of epidemiological surveys should be recognized. As these surveys underpin health policy and allocation of limited resources, misreporting can have far-reaching implications. We report how fraud in a nationwide headache survey occurred and how it was discovered and rectified before it could cause harm. The context was a door-to-door survey to estimate the prevalence and burden of headache disorders in Pakistan. Data were collected from all four provinces of Pakistan by non-medical interviewers and collated centrally. Measures to ensure data integrity were preventative, detective and corrective. We carefully selected and trained the interviewers, set rules of conduct and gave specific warnings regarding the consequences of falsification. We employed two-fold fraud detection methods: comparative data analysis, and face-to-face re-contact with randomly selected participants. When fabrication was detected, data shown to be unreliable were replaced by repeating the survey in new samples according to the original protocol. Comparative analysis of datasets from the regions revealed unfeasible prevalences and gender ratios in one (Multan). Data fabrication was suspected. During a surprise-visit to Multan, of a random sample of addresses selected for verification, all but one had been falsely reported. The data (from 840 cases) were discarded, and the survey repeated with new interviewers. The new sample of 800 cases was demographically and diagnostically consistent with other regions. Fraud in community-based surveys is seldom reported, but no less likely to occur than in other fields of medical research. Measures should be put in place to prevent, detect and, where necessary, correct it. In this instance, had the data from Multan been pooled with those from other regions before analysis, a damaging fraud might have escaped notice.


A nanotechnology-based new approach in the treatment of breast cancer: Biosynthesized silver nanoparticles using Cuminum cyminum L. seed extract.
The present study reports the anticancer activities of Cuminum cyminum L. (Cumin) seed extract, chemically synthetized silver nanoparticles (AgNPs) and biosynthesized silver nanoparticles (Bio-AgNPs) from Cumin seeds on human breast adenocarcinoma cell line (MCF-7) and human breast adenocarcinoma metastatic cell line (AU565). The synthetized nanoparticles were characterized by dynamic light scattering (DLS), UV-visible spectroscopy (UV-Vis), X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR) and Scanning electron microscopy (SEM). The cytotoxic and anticancer effects of AgNPs and Bio-AgNPs were determined by MTT assay. According to the cytotoxicity analysis, Bio-AgNPs appears to be less toxic against J774 macrophage cells than AgNPs since IC50 values were measured as 0.75 and 1.25 μg/ml for AgNPs and Bio-AgNPs, respectively. On the other hand, Bio-AgNPs demonstrated significant inhibitory effects on human breast cancer cells at non-toxic concentrations such as 0.25 and 0.5 μg/ml. However, at increased concentrations, the lethal effects of AgNPs on breast cancer cells were higher than Bio-AgNPs. When cytotoxic and anticancer characteristics of Cumin extract were investigated, it was established that it did not show any inhibitory effect on J774 cells, while killing the half of MCF-7 cells at investigated concentrations. Interestingly, Cumin extract gave rise to no inhibitory effects against AU565 cells. On the other hand, AgNPs and Bio-AgNPs exhibited considerable anticancer activities on both cell lines. The inhibition percentages of AgNPs on MCF-7 and AU565 cell lines were respectively evaluated as 95% and 97% at the highest concentrations applied (12.5 μg/ml). Similarly, we determined that 87.5% and 96% of MCF-7 and AU565 cells were respectively inhibited when they were exposed to the highest concentrations of Bio-AgNPs. Considering relatively toxic-free features of Bio-AgNPs prepared from Cuminum cyminum L. seed extracts, it can be thought that this formulation will be a pioneer in development of nanotechnology-based new anticancer drug for the treatment of breast cancer in near future.


Synergistic Interactions between Abeta, tau, and alpha-synuclein: acceleration of neuropathology and cognitive decline.
Alzheimer's disease (AD), the most prevalent age-related neurodegenerative disorder, is characterized pathologically by the accumulation of beta-amyloid (Abeta) plaques and tau-laden neurofibrillary tangles. Interestingly, up to 50% of AD cases exhibit a third prevalent neuropathology: the aggregation of alpha-synuclein into Lewy bodies. Importantly, the presence of Lewy body pathology in AD is associated with a more aggressive disease course and accelerated cognitive dysfunction. Thus, Abeta, tau, and alpha-synuclein may interact synergistically to promote the accumulation of each other. In this study, we used a genetic approach to generate a model that exhibits the combined pathologies of AD and dementia with Lewy bodies (DLB). To achieve this goal, we introduced a mutant human alpha-synuclein transgene into 3xTg-AD mice. As occurs in human disease, transgenic mice that develop both DLB and AD pathologies (DLB-AD mice) exhibit accelerated cognitive decline associated with a dramatic enhancement of Abeta, tau, and alpha-synuclein pathologies. Our findings also provide additional evidence that the accumulation of alpha-synuclein alone can significantly disrupt cognition. Together, our data support the notion that Abeta, tau, and alpha-synuclein interact in vivo to promote the aggregation and accumulation of each other and accelerate cognitive dysfunction.


Beta-C-mannosides as selectin inhibitors.
Potential E- and P-selectin inhibitors were synthesized to explore a hydrophobic area on the E-selectin surface and the PSGL-1 protein binding site on the P-selectin surface that was recently defined by crystallography. Three series of mannose-based compounds (libraries A, B, and C) were synthesized using solution phase parallel synthesis. Biological evaluation of these compounds was done using two ELISA-based assays and transferred NOE (trNOE) experiments. Some of the compounds showed better activity than sLe(x) in the P-selectin assay.


Impact of tachycardia and sympathetic stimulation by cold pressor test on cardiac diastology and arterial function in elderly females.
Abnormal vascular-ventricular coupling has been suggested to contribute to heart failure with preserved ejection fraction in elderly females. Failure to increase stroke volume (SV) during exercise occurs in parallel with dynamic changes in arterial physiology leading to increased afterload. Such adverse vascular reactivity during stress may reflect either sympathoexcitation or be due to tachycardia. We hypothesized that afterload elevation induces SV failure by transiently attenuating left ventricular relaxation, a phenomenon described in animal research. The respective roles of tachycardia and sympathoexcitation were investigated in n = 28 elderly females (70 ± 4 yr) carrying permanent pacemakers. At rest, during atrial tachycardia pacing (ATP; 100 min(-1)) and during cold pressor test (hand immersed in ice water), we performed Doppler echocardiography (maximal untwist rate analyzed by speckle tracking imaging of rotational mechanics) and arterial tonometry (arterial stiffness estimated as augmentation index). Estimation of arterial compliance was based on an exponential relationship between arterial pressure and volume. We found that ATP produced central hypovolemia and a reduction in SV which was larger in patients with stiffer arteries (higher augmentation index). There was an associated adverse response of arterial compliance and vascular resistance during ATP and cold pressor test, causing an overall increase in afterload, but nonetheless enhanced maximal rate of untwist and no evidence of afterload-dependent failure of relaxation. In conclusion, tachycardia and cold provocation in elderly females produces greater vascular reactivity and SV failure in the presence of arterial stiffening, but SV failure does not arise secondary to afterload-dependent attenuation of relaxation.


Fatal blunt aortic injuries: a review of 242 autopsy cases.
To characterize fatal blunt aortic injury (BAI). A retrospective chart review of 242 cases of fatal BAI in patients who underwent an autopsy at our institution between 1984 and 1997 was performed. Comparisons were made for statistical differences using the z-test. Two hundred forty-two cases of fatal BAI were reviewed, making this the largest BAI autopsy study to date. Mechanisms of BAI included driver/passenger in motor vehicle crash (MVC) (68%), pedestrian versus MVC (17%), and motorcycle crash (8%). When comparing the mechanisms in the time period 1984 to 1988 to the time period 1989 to 1997, only the pedestrian versus MVC mechanism was significantly different (12% vs. 23%, p < 0.05). MVC direction of impact included head-on (45%), lateral (35%), and complex (20%). Two thirds of the victims sustained head injuries, rib fractures, and/or hepatic trauma. Only 58% of the victims had the classic isthmus laceration. There was one preventable death secondary to delay in diagnosis. BAI is not limited to frontal impact crashes; there should be a high index of suspicion of BAI in lateral impact crashes as well as pedestrian versus MVC mechanisms. Nonisthmus and complex aortic lacerations are common in fatal BAI. Finally, BAI is a highly lethal injury with few preventable deaths in this series.


3-D ultrastructural distribution of collagen in human placental villi at term in relation to vascular tree.
In order to understand the 3-D distribution of collagen in relation to vascularization, chorionic villi of human placentae, belonging to normal pregnancies at term, were studied by scanning electron microscopy (SEM) after alkali maceration techniques, and by transmission electron microscopy (TEM). The villous tree appeared made of an uninterrupted structure of collagen fibres. The collagen fibres connected the chorionic villi axis with their basal plates and organised differently according to the various levels of villous branching. The collagen of stem villi showed copious fibres. The external fibres (facing the villous surface) were arranged mainly longitudinally. The central core of the villi (inner fibres) were arranged concentrically around the wall of the fetal vessels. Both external and internal fibres formed stratified lamellae or small parallel bundles. The inner core of stem villi showed small holes housing capillary spaces. Mature intermediate and terminal villi showed a scarce amount of collagen arranged in thin concentric layer within the villous core, surrounding numerous dilated capillary and sinusoid spaces.These observations demonstrated that the extracellular matrix of human chorionic villi is highly compartmentalised and shows a variable structural 3-D distribution depending on the branching level of the villous tree, such a distribution ensures the most favourable microenvironment for feto-maternal exchanges and it is likely able to provide a modulated support to the developing chorionic fetal vessels and trophoblastic layer as well.


The effects of the broadband UVA radiation on myeloid leukemia cells: the possible role of protein kinase C in mediation of UVA-induced effects.
We examined the effects of broadband UVA radiation (320-400 nm) on a rat myeloid leukemia cell line-chloroma (ChL). A Phillips face tanner model HB 171/A was used as a light source. Chloroma were irradiated through a 5 mm thick glass filter that cut off all of the UVB contamination. The irradiances were measured, from 250 to 400 nm, with a well-characterized and calibrated double-grating spectroradiometer Optronic 742. The overall uncertainty of dose evaluation was estimated to be +/-15% (2 sigma). The cells were irradiated with UVA doses of 4 and 8 J/cm2 and cultured thereafter for 24 h. After this period of time, a marked decline up to 50% was observed in cell proliferation in UVA-irradiated ChL cultures. The cell proliferation decline was found to be caused by simultaneously occurring G2/M phase cell cycle arrest and apoptosis in part of the UVA-irradiated ChL population. Concomitantly, with the decline in cell proliferation, an increase was observed in the expression of the major histocompatibility (MHC) class I and II antigens. Because protein kinase C (PKC) is known to regulate cell proliferation, apoptosis and expression of MHC antigens, and because UVA was shown to regulate PKC activity/expression, we therefore examined whether UVA irradiation has any effect on the expression of isozymes of PKC. Western blots revealed that ChL express alpha, beta I, delta, epsilon, eta, and zeta/iota isozymes of PKC and that expression of all isozymes declined 24 h after UVA irradiation (8 J/cm2). Finally, PKC activation in ChL by exposure to phorbol ester caused cell cycle arrest in G1 phase but did not induce apoptosis. This suggests that the previously shown UVA-induced PKC activation in ChL might be responsible for the induction of MHC antigens but the simultaneously observed ChL apoptosis is likely to be mediated by PKC down-regulation. All together, our results suggest that UVA, at irradiance levels that resemble the outdoor exposure, may have profound effects on the immune-related properties of leukocytes. Thus, we speculate that in vivo the immune functions of leukocytes passing through dermal capillaries might be altered by exposure to solar UVA radiation.


[Ovarian cancer 5 years after hysterectomy with adnexectomy for a benign lesion. Apropos of a case].
The rare and strange evolution of an ovarian tumour in a woman of 58 years of age. Extraordinary in its method of presentation and interesting because of the way it was decided to treat it and because of the theories concerning its physiopathology. A pelvic mass that appeared 5 years after hysterectomy with bilateral salpingo-oophorectomy for a fibroid proved to be an ovarian cancer. The diagnosis was difficult and the treatment very complicated.


Transmembrane topology of FRO2, a ferric chelate reductase from Arabidopsis thaliana.
Iron uptake in Arabidopsis thaliana is mediated by ferric chelate reductase FRO2, a transmembrane protein belonging to the flavocytochrome b family. There is no high resolution structural information available for any member of this family. We have determined the transmembrane topology of FRO2 experimentally using the alkaline phosphatase fusion method. The resulting topology is different from that obtained by theoretical predictions and contains 8 transmembrane helices, 4 of which build up the highly conserved core of the protein. This core is present in the entire flavocytochrome b family. The large water soluble domain of FRO2, which contains NADPH, FAD and oxidoreductase sequence motifs, was located on the inside of the membrane.


Planteome 2024 Update: Reference Ontologies and Knowledgebase for Plant Biology.
The Planteome project (https://planteome.org/) provides a suite of reference and crop-specific ontologies and an integrated knowledgebase of plant genomics data. The plant genomics data in the Planteome has been obtained through manual and automated curation and sourced from more than 40 partner databases and resources. Here, we report on updates to the Planteome reference ontologies, namely, the Plant Ontology (PO), Trait Ontology (TO), the Plant Experimental Conditions Ontology (PECO), and integration of species/crop-specific vocabularies from our partners, the Crop Ontology (CO) into the TO ontology graph. Currently, 11 CO vocabularies are integrated into the Planteome with the addition of yam, sorghum, and potato since 2018. In addition, the size of the annotation database has increased by 34%, and the number of bioentities (genes, proteins, etc.) from 125 plant taxa has increased by 72%. We developed new tools to facilitate user requests and improvements to the CO vocabularies, and to allow fast searching and browsing of PO terms and definitions. These enhancements and future changes to automate the TO-CO mappings and knowledge discovery tools ensure that the Planteome will continue to be a valuable resource for plant biology.


Structural changes in the Mn4Ca cluster and the mechanism of photosynthetic water splitting.
Photosynthetic water oxidation, where water is oxidized to dioxygen, is a fundamental chemical reaction that sustains the biosphere. This reaction is catalyzed by a Mn4Ca complex in the photosystem II (PS II) oxygen-evolving complex (OEC): a multiprotein assembly embedded in the thylakoid membranes of green plants, cyanobacteria, and algae. The mechanism of photosynthetic water oxidation by the Mn4Ca cluster in photosystem II is the subject of much debate, although lacking structural characterization of the catalytic intermediates. Biosynthetically exchanged Ca/Sr-PS II preparations and x-ray spectroscopy, including extended x-ray absorption fine structure (EXAFS), allowed us to monitor Mn-Mn and Ca(Sr)-Mn distances in the four intermediate S states, S0 through S3, of the catalytic cycle that couples the one-electron photochemistry occurring at the PS II reaction center with the four-electron water-oxidation chemistry taking place at the Mn4Ca(Sr) cluster. We have detected significant changes in the structure of the complex, especially in the Mn-Mn and Ca(Sr)-Mn distances, on the S2-to-S3 and S3-to-S0 transitions. These results implicate the involvement of at least one common bridging oxygen atom between the Mn-Mn and Mn-Ca(Sr) atoms in the O-O bond formation. Because PS II cannot advance beyond the S2 state in preparations that lack Ca(Sr), these results show that Ca(Sr) is one of the critical components in the mechanism of the enzyme. The results also show that Ca is not just a spectator atom involved in providing a structural framework, but is actively involved in the mechanism of water oxidation and represents a rare example of a catalytically active Ca cofactor.


[Alpha 2-antiplasmin in bovine plasma].
Bovine and human blood plasma contains alpha 2-antiplasmin which possesses affinity to lysin-binding sites in plasmin and inhibits the human plasmin. Its isolation was conducted for two stages: separation of plasminogen on lysin-cellulose, fractionation by ammonium sulphate, desalination on molselector G-25, chromatography on DEAE-Sephadex A-50 and affinity chromatography on plasmin-sepharose with the blocked active site.


Influence of vitamin E and ICRF-187 on chronic doxorubicin cardiotoxicity in miniature swine.
Studies were made of the potential of vitamin E and ICRF-187 to protect against the cardiotoxicity resulting from chronic administration of doxorubicin. Miniature swine (19 to 40 kg) received six injections of doxorubicin (1.6 mg/kg) at 3-week intervals (total dose, 9.6 mg/kg), either alone or concurrently with vitamin E (5000 IU/day for 4 days and 1000 units/day for the next 17 days). In a second study, miniature swine received six injections of doxorubicin (2.4 mg/kg) at 3-week intervals (total dose 14.4 mg/kg), either alone or 30 minutes after 12.5 mg of ICRF-187/kg (intraperitoneally). All animals were sacrificed 3 weeks after the last injection. The frequency and extent of myocardial lesions (vacuolization and myofibrillar loss) were scored on a scale of 0 to 4+. Such lesions were noted in eight of nine pigs given 9.6 mg/kg of doxorubicin alone and in all pigs receiving doxorubicin and vitamin E; however, the severity of the lesions was decreased in the latter animals (average score 1.0, compared with 1.8 in those receiving doxorubicin alone). All swine receiving 14.4 mg/kg of doxorubicin alone developed myocardial lesions (average score, 2.7); these lesions were severe (3+) in four of the animals. In contrast, cardiac lesions were absent in two and minimal (average score, 0.7) in five of the seven animals given 14.4 mg/kg of doxorubicin in combination with ICRF-187.


Influence of ketamine, propofol or isoflurane on intraocular pressure, heart rate and blood pressure in healthy dogs premedicated with medetomidine and midazolam.
According to the findings of several studies, sedatives and anaesthetics have different effects on the functioning of the cardiovascular system and intraocular pressure (IOP). For accurate diagnosis, treatment and surgery with minimal complications, it is necessary to be aware of the effects of sedatives and anaesthetics on the cardiovascular system and IOP. The aim of this study was to evaluate the effects of sedatives (medetomidine and midazolam) and anaesthetics (ketamine, propofol and isoflurane) on IOP, heart rate (HR) and blood pressure in dogs. In this study, 10 dogs participated in three treatments using a randomised cross-over design, with a 1-week washout period between each treatment. Dogs in all treatments were premedicated with medetomidine and midazolam. Anaesthesia was induced using ketamine, propofol, or isoflurane and maintained for 60 min with the appropriate doses of each drug. The cardiovascular variables (heart rate, and systolic, diastolic and mean arterial pressures) and IOP were measured at different timepoints: before premedication (baseline values, T-Bas), 15 min after medetomidine administration (T-Med), 20 min after midazolam administration (T-Mid) and at 15 (T-15), 30 (T-30), 45 (T-45) and 60 (T-60) min after anaesthesia induction. Medetomidine significantly reduced the IOP and HR and did not significantly change the mean arterial pressure (MAP). Midazolam significantly reduced the IOP while did not significantly change the HR and MAP. Ketamine and isoflurane significantly increased the IOP and HR while did not significantly change the MAP. Propofol significantly increased the HR, but did not cause significant changes in IOP and MAP. Considering that anaesthetics are typically administered in conjunction with pre-anaesthetic drugs, the increases in IOP induced by ketamine and isoflurane are not important, as the IOP did not exceed the baseline values. However, further studies are required to investigate these effects in patients with elevated IOP.


Torsional injuries of the lower limb: an analysis of the frictional torque between different types of football turf and the shoe outsole.
Football turf is increasingly used in European soccer competition. Little is known on the rotational torque that players experience on these fields. High rotational torques between the shoe outsole and the sports surface has been correlated with torsional injuries of the lower limb and knee. To evaluate the effect of six parameters that could influence the rotational torque between the shoe outsole and the latest generation football turf. Controlled laboratory study. A testing apparatus was constructed to measure the peak torque generated during a controlled rotation of the foot. Six parameters that could potentially influence the frictional forces, were considered: (1) the sports surface, (2) the shoe outsole cleat design, (3) the weather conditions, (4) the weight, (5) the presence of an impact and (6) the direction of rotation. The football turf without infill showed significantly lower frictional torques than natural grass whereas a football turf with sand/rubber infill had significantly higher torques. Blades were associated with significantly higher torques than studs on natural grass and on one football turf with sand/rubber infill. Dry weather was associated with higher torques only for the football turf without infill. The torque increased linearly and significantly with an increasing vertical load. The rotational torque increased significantly following an impact. Torques on external rotational movements were significantly higher with blades. Important differences in rotational torques are found and could be seen as potential risk factors for torsional injuries of the lower limb.


Protein kinase Cbeta inhibition attenuates osteopontin expression, macrophage recruitment, and tubulointerstitial injury in advanced experimental diabetic nephropathy.
Tubulointerstitial macrophage accumulation is an important marker of prognosis that correlates closely with declining renal function in a range of human and experimental diseases, including diabetic nephropathy. These inflammatory cells are rich in the profibrotic growth factor TGF-beta such that their presence in areas of injury is frequently associated with tissue fibrosis. The migration of macrophages occurs in response to the site-specific production of chemokines, with osteopontin closely associated with their trafficking into the tubulointerstitium of the kidney. Although cell culture studies indicate that protein kinase C (PKC) mediates the expression of osteopontin, its role in the in vivo setting is unknown. Accordingly, Ren-2 control and diabetic rats that were treated with or without the specific PKC-beta isoform inhibitor ruboxistaurin (10 mg/kg per d) were examined. After 12 wk, diabetic rats showed increases in osteopontin expression in tubular epithelial cells of the cortex in association with macrophage infiltration, interstitial fibrosis, and activity of TGF-beta as indicated by the expression of its receptor activated protein phospho-Smad2 (P < 0.05 for all parameters). Ruboxistaurin treatment significantly attenuated these parameters (P < 0.05) in diabetic rats without affecting either BP or glycemic control. These findings suggest that osteopontin and macrophage accumulation may play a role in the tubulointerstitial injury in diabetic nephropathy and that inhibition of osteopontin expression may be one of the mechanisms by which inhibition of the beta-isoform of PKC confers a renoprotective effect.


Crystal structure of unligated guanylate kinase from yeast reveals GMP-induced conformational changes.
The crystal structure of guanylate kinase (GK) from yeast (Saccharomyces cerevisiae) with a non-acetylated N terminus has been determined in its unligated form (apo-GK) as well as in complex with GMP (GK.GMP). The structure of apo-GK was solved with multiwavelength anomalous diffraction data and refined to an R-factor of 0.164 (R(free)=0.199) at 2.3 A resolution. The structure of GK.GMP was determined using the crystal structure of GK with an acetylated N terminus as the search model and refined to an R-factor of 0.156 (R(free)=0.245) at 1.9 A. GK belongs to the family of nucleoside monophosphate (NMP) kinases and catalyzes the reversible phosphoryl transfer from ATP to GMP. Like other NMP kinases, GK consists of three dynamic domains: the CORE, LID, and NMP-binding domains. Dramatic movements of the GMP-binding domain and smaller but significant movements of the LID domain have been revealed by comparing the structures of apo-GK and GK.GMP. apo-GK has a much more open conformation than the GK.GMP complex. Systematic analysis of the domain movements using the program DynDom shows that the large movements of the GMP-binding domain involve a rotation around an effective hinge axis approximately parallel with helix 3, which connects the GMP-binding and CORE domains. The C-terminal portion of helix 3, which connects to the CORE domain, has strikingly higher temperature factors in GK.GMP than in apo-GK, indicating that these residues become more mobile upon GMP binding. The results suggest that helix 3 plays an important role in domain movement. Unlike the GMP-binding domain, which moves toward the active center of the enzyme upon GMP binding, the LID domain moves away from the active center and makes the presumed ATP-binding site more open. Therefore, the LID domain movement may facilitate the binding of MgATP. The structure of the recombinant GK.GMP complex superimposes very well with that of the native GK.GMP complex, indicating that N-terminal acetylation does not have significant impact on the three-dimensional structure of GK.


Grade III Liver Laceration in a Female Volleyball Player.
A 20-year-old female collegiate volleyball athlete (body mass, 74.8 kg; height, 177.8 cm), with no previous history of abdominal injury, dove for a ball during a match and was struck in the anterolateral abdominal region by a teammate's knee. She experienced a solar plexus spasm, shortness of breath, and nausea. On-site evaluation revealed sharp pain in the right upper quadrant and epigastric area, pallor, continued nausea, and excessive sweating. Upon arrival to the emergency department, an abdominal/pelvic computed tomography (CT) scan with intravenous contrast showed a small amount of fluid along the posteromedial surface of the liver indicating a subcapsular hematoma. A trace amount of free fluid in the right paracolic gutter and pelvis was also noted, suggesting a hemoperitoneum. The athlete was diagnosed with a grade III laceration (> 3 cm parenchymal depth) on the right lobe of the liver. She was hemodynamically stable and vital sign values were normal ~2.5 hours postinjury (blood pressure, 118/76 mm Hg; heart rate, 68 beats per minute; respiratory rate, 16 breaths per minute; tympanic temperature, 36.3°C). The athlete was admitted into the intensive care unit for observation, and the attending physician decided on a conservative, nonoperative approach to treatment. Forty-eight hours postinjury, a second abdominal/pelvic CT scan was ordered and revealed no increased bleeding or extensive damage to the liver. Two days later, after being discharged from the hospital, she was prohibited from practicing until a third abdominal/pelvic CT scan was performed and showed no residual injury to the liver (5 weeks postinjury). The athlete returned to play 6 weeks postinjury without complications; however, she continued performing rehabilitation focusing on cardiorespiratory endurance.


Placing Racial Classification in Context.
This article extends previous research on place-based patterns of racial categorization by linking it to sociological theory that posits subnational variation in cultural schemas and applying regression techniques that allow for spatial variation in model estimates. We use data from a U.S. restricted-use geocoded longitudinal survey to predict racial classification as a function of both individual and county characteristics. We first estimate national average associations, then turn to spatial-regime models and geographically weighted regression to explore how these relationships vary across the country. We find that individual characteristics matter most for classification as "Black," while contextual characteristics are important predictors of classification as "White" or "Other," but some predictors also vary across space, as expected. These results affirm the importance of place in defining racial boundaries and suggest that U.S. racial schemas operate at different spatial scales, with some being national in scope while others are more locally situated.


Massage for promoting growth and development of preterm and/or low birth-weight infants.
It has been argued that infants in Neonatal Intensive Care Units are subject both to a highly stressful environment - continuous, high-intensity noise and bright light - and to a lack of the tactile stimulation that they would otherwise experience in the womb or in general mothering care. As massage seems to both decrease stress and provide tactile stimulation, it has been recommended as an intervention to promote growth and development of preterm and low-birth weight infants. To determine whether preterm and / or low birth-weight infants exposed to massage experience improved weight gain and earlier discharge compared to infants receiving standard care; to determine whether massage has any other beneficial or harmful effects on this population. Databases were searched using the term 'massage', 'touch' or 'tactile stimulation' with 'infant - newborn', 'infant - premature' and 'infant - low birth weight'. The main databases searched were those of the Cochrane Collaboration Field in Complementary Medicine and the Neonatal Collaborative Review Group. Randomised trials in which infants with gestational age at birth < 37 weeks or weight at birth < 2500g received systematic tactile stimulation by human hands. At least one outcome assessing weight gain, length of stay, behaviour or development must be reported. Data extracted from each trial were baseline characteristics of sample, weight gain, length of stay and behavioural and developmental outcomes. Physiological and biochemical outcomes were not recorded. Data were extracted by three reviewers independently. Statistical analysis was conducted using the standard Cochrane Collaboration methods. Massage interventions improved daily weight gain by 5g (95% CI 3.5, 6.7g). There is no evidence that gentle, still touch is of benefit (increase in daily weight gain -0.2g; 95% CI -2.4, 1.9g). Massage interventions also appeared to reduce length of stay by 4.6 days (95% CI 2.6, 6.6) though there are methodological concerns about the blinding of this outcome. There was also some evidence that massage interventions have a slight, positive effect on postnatal complications and weight at 4 - 6 months. However, serious concerns about the methodological quality of the included studies, particularly with respect to selective reporting of outcomes, weaken credibility in these findings. Evidence that massage for preterm infants is of benefit for developmental outcomes is weak and does not warrant wider use of preterm infant massage. Where massage is currently provided by nurses, consideration should be given as to whether this is a cost-effective use of time. Future research should assess the effects of massage interventions on clinical outcome measures, such as medical complications or length of stay, and on process-of-care outcomes, such as care-giver or parental satisfaction.


Annexin A3 as a potential target for immunotherapy of liver cancer stem-like cells.
Cancer stem-like cells/cancer-initiating cells (CSCs/CICs) are considered to represent a small population of cancer cells that is resistant to conventional cancer treatments and responsible for tumor recurrence and metastasis. The aim of this study was to establish CSC/CIC-targeting immunotherapy. In this study, we found that Annexin A3 (ANXA3) was preferentially expressed in CSCs/CICs derived from hepatocellular carcinoma (HCC) cells compared to non-CSCs/CICs. In HCC samples, high levels of ANXA3 correlated with expansion of CD133(+) tumor cells representing CSCs/CICs in HCC; the combination of high levels of ANXA3 and CD133 was associated with progression of HCC. Overexpression of ANXA3 increased the proportion of CD133(+) cells, enhancing their tumorigenicity. On the contrary, knockdown of ANXA3 decreased CD133(+) cells and inhibited tumorigenicity. The mechanistic study revealed that ANXA3-mediated maintenance of HCC CSCs/CICs activity was likely involved with the HIF1A/Notch pathway. Using ANXA3 as a target, ANXA3-transfected dendritic cells could induce more functionally active T cells and these effector T cells could superiorly kill CD133(+) HCC CSCs/CICs in vitro and in vivo. Taken together, our findings suggest that ANXA3 plays a role in HCC CSC/CIC maintenance, and that ANXA3 may represent a potential CSC/CIC-specific therapeutic target for improving the treatment of HCC.


Oral Nonviral Gene Delivery for Chronic Protein Replacement Therapy.
Efficient nonviral oral gene delivery offers an attractive modality for chronic protein replacement therapy. Herein, the oral delivery of insulin gene is reported by a nonviral vector comprising a copolymer with a high degree of substitution of branched polyethylenimine on chitosan (CS-g-bPEI). Protecting the plasmid from gastric acidic degradation and facilitating transport across the gut epithelium, the CS-g-bPEI/insulin plasmid DNA nanoparticles (NPs) can achieve systemic transgene expression for days. A single dose of orally administered NPs (600 µg plasmid insulin (pINS)) to diabetic mice can protect the animals from hyperglycemia for more than 10 d. Three repeated administrations spaced over a 10 d interval produce similar glucose-lowering results with no hepatotoxicity detected. Positron-emission-tomography and computed-tomography images also confirm the glucose utilization by muscle cells. While this work suggests the feasibility of basal therapy for diabetes mellitus, its significance lies in the demonstration of a nonviral oral gene delivery system that can impact chronic protein replacement therapy and DNA vaccination.


Reporting and Guidelines in Propensity Score Analysis: A Systematic Review of Cancer and Cancer Surgical Studies.
: Propensity score (PS) analysis is increasingly being used in observational studies, especially in some cancer studies where random assignment is not feasible. This systematic review evaluates the use and reporting quality of PS analysis in oncology studies. : We searched PubMed to identify the use of PS methods in cancer studies (CS) and cancer surgical studies (CSS) in major medical, cancer, and surgical journals over time and critically evaluated 33 CS published in top medical and cancer journals in 2014 and 2015 and 306 CSS published up to November 26, 2015, without earlier date limits. The quality of reporting in PS analysis was evaluated. It was also compared over time and among journals with differing impact factors. All statistical tests were two-sided. More than 50% of the publications with PS analysis from the past decade occurred within the past two years. Of the studies critically evaluated, a considerable proportion did not clearly provide the variables used to estimate PS (CS 12.1%, CSS 8.8%), incorrectly included non baseline variables (CS 3.4%, CSS 9.3%), neglected the comparison of baseline characteristics (CS 21.9%, CSS 15.6%), or did not report the matching algorithm utilized (CS 19.0%, CSS 36.1%). In CSS, the reporting of the matching algorithm improved in 2014 and 2015 ( P = .04), and the reporting of variables used to estimate PS was better in top surgery journals ( P = .008). However, there were no statistically significant differences for the inclusion of non baseline variables and reporting of comparability of baseline characteristics. The use of PS in cancer studies has dramatically increased recently, but there is substantial room for improvement in the quality of reporting even in top journals. Herein we have proposed reporting guidelines for PS analyses that are broadly applicable to different areas of medical research that will allow better evaluation and comparison across studies applying this approach.


Menthol Cigarettes, Tobacco Dependence, and Smoking Persistence: The Need to Examine Enhanced Cognitive Functioning as a Neuropsychological Mechanism.
Despite the overall decline in the prevalence of cigarette use in the United States, menthol cigarette use among smokers is rising, and evidence shows that it may lead to more detrimental effects on public health than regular cigarette use. One of the mechanisms by which nicotine sustains tobacco use and dependence is due to its cognitive enhancing properties, and basic science literature suggests that menthol may also enhance nicotine's acute effect on cognition. The purpose of this review is to suggest that the cognitive enhancing effects of menthol may be a potentially important neuropsychological mechanism that has yet to be examined. In this narrative review, we provide an overview of basic science studies examining neurobiological and cognitive effects of menthol and menthol cigarette smoking. We also review studies examining menthol essential oils among humans that indicate menthol alone has acute cognitive enhancing properties. Finally, we present factors influencing the rising prevalence of menthol cigarette use among smokers and the importance of this gap in the literature to improve public health and smoking cessation treatment. Despite the compelling evidence for menthol's acute cognitive enhancing and reinforcing effects, this mechanism for sustaining tobacco dependence and cigarette use has yet to be examined and validated among humans. On the basis of the basic science evidence for menthol's neurobiological effects on nicotinic receptors and neurotransmitters, perhaps clarifying menthol's effect on cognitive performance can help to elucidate the complicated literature examining menthol and tobacco dependence. Menthol cigarette use has continued to be a topic of debate among researchers and policy makers, because of its implications for understanding menthol's contribution to nicotine dependence and smoking persistence, as well as its continued use as a prevalent flavoring in tobacco and nicotine products in the United States and internationally. As international tobacco regulation policies have begun to target menthol cigarettes, research studies need to examine how flavoring additives, specifically menthol, may acutely influence neurobiological and cognitive functioning as a potential mechanism of sustained smoking behavior to develop more effective treatments.


Long-term neurologic outcomes after traumatic brain injury.
To determine the relations between traumatic brain injury (TBI) and several neurologic outcomes 6 months or more after TBI. Not applicable. Systematic review of the published, peer-reviewed literature. Not applicable. We identified 75 studies that examined the relations between TBI and neurologic outcomes. Unprovoked seizures are causally related to penetrating TBI as well as to moderate and severe TBI. There was only limited evidence of an association between seizures and mild TBI. Dementia of the Alzheimer's type (DAT) was associated with moderate and severe TBI, but not with mild TBI unless there was loss of consciousness (LOC); the evidence for the latter was limited. Parkinsonism was associated with moderate and severe TBI, but there was only modest evidence of a link with mild TBI without LOC. Dementia pugilistica was associated with professional boxing. There was insufficient evidence to support an association between TBI and both multiple sclerosis and amyotrophic lateral sclerosis. TBI appeared to produce a host of postconcussive symptoms (eg, memory problems, dizziness, and irritability). Moderate and severe TBI were associated with endocrine problems such as hypopituitarism and growth hormone deficiency and possibly with diabetes insipidus. There was only limited evidence of an association between mild TBI and the development of ocular/visual motor deterioration. TBI is strongly associated with several neurologic disorders 6 months or more after injury. Clinicians caring for TBI patients should monitor them closely for the development of these disorders. While some of these disorders can be treated after they arise (eg, seizures), a greater public health benefit would be achieved by preventing them before they develop. Research efforts to develop therapies aimed at secondary prevention are currently underway.


[Effectiveness of treating periodontitis in patients with thyroid dysfunction].
Efficiency of some drugs in the treatment of periodontitis in combination with corrective treatment of thyroid function was evaluated in 70 patients with hypo- and hyperthyrosis with different initial level of nonspecific resistance. The therapeutic complex including drugs commonly used in the treatment of periodontitis and irrigation of the periodontium with lithium chloride and chlorohexidine solutions was highly effective in patients with thyroid dysfunction and relatively favorable status of nonspecific resistance of the organism. In patients with hypo- and hyperthyrosis with poor nonspecific resistance the best effect in the treatment of periodontitis was attained with potassium orotate as an immunomodulator and lithium chloride.


Genome-wide linkage analyses of systolic blood pressure using highly discordant siblings.
Elevated blood pressure is a risk factor for cardiovascular, cerebrovascular, and renal diseases. Complex mechanisms of blood pressure regulation pose a challenge to identifying genetic factors that influence interindividual blood pressure variation in the population at large. We performed a genome-wide linkage analysis of systolic blood pressure in humans using an efficient, highly discordant, full-sibling design. We identified 4 regions of the human genome that show statistical significant linkage to genes that influence interindividual systolic blood pressure variation (2p22.1 to 2p21, 5q33.3 to 5q34, 6q23.1 to 6q24.1, and 15q25.1 to 15q26.1). These regions contain a number of candidate genes that are involved in physiological mechanisms of blood pressure regulation. These results provide both novel information about genome regions in humans that influence interindividual blood pressure variation and a basis for identifying the contributing genes. Identification of the functional mutations in these genes may uncover novel mechanisms for blood pressure regulation and suggest new therapies and prevention strategies.


LeucoPatch system for the management of hard-to-heal diabetic foot ulcers in the UK, Denmark, and Sweden: an observer-masked, randomised controlled trial.
The LeucoPatch device uses bedside centrifugation without additional reagents to generate a disc comprising autologous leucocytes, platelets, and fibrin, which is applied to the surface of the wound. We aimed to test the effectiveness of LeucoPatch on the healing of hard-to-heal foot ulcers in people with diabetes. This was a multicentre, international, observer-masked, randomised controlled trial of people with diabetes and a hard-to-heal foot ulcer done in 32 specialist diabetic foot clinics in three countries (UK, Denmark, and Sweden). After a 4-week run-in period, those with a reduction in ulcer area of less than 50% were randomly allocated (1:1) by computer-generated, web-based randomisation (block sizes of two, four, and six) to either prespecified good standard care alone or care plus weekly application of LeucoPatch. The primary outcome was the proportion of ulcers that healed within 20 weeks assessed in the intention-to-treat population (all participants with post-randomisation data collected), defined as complete epithelialisation (confirmed by an observer who was masked to randomisation group), and remained healed for 4 weeks. This trial is registered with the ISRCTN registry, number 27665670, and ClinicalTrials.gov, number NCT02224742. Between Aug 30, 2013, and May 3, 2017, 269 participants were randomly allocated to receive treatment (137 to receive standard care and 132 to receive LeucoPatch). The mean age was 61·9 years (SD 11·6), 217 (82%) were men, and 222 (83%) had type 2 diabetes. In the LeucoPatch group, 45 (34%) of 132 ulcers healed within 20 weeks versus 29 (22%) of 134 ulcers in the standard care group (odds ratio 1·58, 96% CI 1·04-2·40; p=0·0235) by intention-to-treat analysis. Time to healing was shorter in the LeucoPatch group (p=0·0246) than in the standard care group. No difference in adverse events was seen between the groups. The most common serious adverse event (SAE) was diabetic foot infection (24 events in the LeucoPatch group [24% of all SAEs] and 20 in the standard care group [27% of all SAEs]. There were no device-related adverse events. The use of LeucoPatch is associated with significant enhancement of healing of hard-to-heal foot ulcers in people with diabetes. Reapplix ApS.


Using pulmonary imaging to move chronic obstructive pulmonary disease beyond FEV1.
FEV1, measured using spirometry, provides a straightforward, widely available, and inexpensive global measurement of airflow limitation and lung function. For decades, FEV1 has remained the main intermediate endpoint used in research studies and for the development of new chronic obstructive pulmonary disease (COPD) therapies. Not surprisingly, treatments that acutely improve FEV1 dominate as COPD therapies. However, in patients with COPD, the relationship of FEV1 with symptoms and outcomes such as exacerbations and mortality is weak, and, importantly, FEV1 does not take into account the heterogeneity of COPD or its different phenotypes. Thoracic imaging provides a way to quantify airway remodeling, emphysematous destruction, regional ventilation abnormalities (ventilation defects), and gas trapping in ex-smokers in whom FEV1 may be normal and in patients with COPD with very modest lung function deterioration. In individual patients and in COPD cohort studies, thoracic imaging using X-ray computed tomography, and magnetic resonance imaging (conventional (1)H as well as hyperpolarized noble gases such as (129)Xe, (3)He, and inhaled O2 and (19)F) can be used to directly visualize the structural and functional consequences of COPD and thus provide a clearer picture of COPD mechanisms, disease progression, and response to therapy. We briefly describe pulmonary imaging methods that provide a way to visualize and quantify, with high spatial and temporal resolution, regional ventilation abnormalities, gas trapping, emphysema, and airway remodeling in COPD. Finally, we discuss the implications of recent imaging findings and their impact on future biomarker and therapy research aimed at improving COPD outcomes.


How much do combined affective and cognitive impairments worsen rehabilitation outcomes after hip fracture ?
To investigate the association between isolated and combined affective and cognitive impairments with functional outcomes and discharge destination in older patients admitted to rehabilitation after a hip fracture. Prospective study in 612 community-dwelling patients aged 65 years and over, admitted to rehabilitation after surgery for hip fracture. Information on socio-demographics, medical, functional, affective, and cognitive status was systematically collected at admission. Functional status, length of stay and destination were assessed at discharge. Functional improvement was defined as any gain on the Barthel Index score between admission and discharge from rehabilitation. At admission, 8.2% of the patients had isolated affective impairment, 27.5% had cognitive impairment only, and 7.5% had combined impairments. Rate of functional improvement steadily decreased from 91.2% in patients with no cognitive nor affective impairment to 73.8% in those with combined impairments. Compared to patients without any impairment, those with combined impairments had lower odds of functional improvement, even after adjustment for age, gender, health and functional status at admission (adjOR: 0.40; 95%CI: 0.16-1.0; p = .049). The proportion of patients discharged back home gradually decreased from 82.8% among patients without any impairment to only 45.6% in patients with combined impairments. In multivariate analysis, the odds of returning home remained significantly reduced in these latter patients (adjOR: 0.31; 95%CI:0.15-0.66; p = .002). Affective and cognitive impairments had both independent, and synergistic negative association with functional outcome and discharge destination in patients admitted to rehabilitation after a hip fracture. Nevertheless, patients with combined affective and cognitive impairments still achieved significant functional improvement, even though its magnitude was reduced. Further studies should investigate whether these patients would benefit from better targeted, longer, or more intensive rehabilitation interventions to optimize their functional recovery.


Evidence for novel tomato seed allergens: IgE-reactive legumin and vicilin proteins identified by multidimensional protein fractionation-mass spectrometry and in silico epitope modeling.
Tomato fruit and seed allergens were detected by IgE-immunoblotting using sera from 18 adult tomato-sensitized patients selected based on a positive history skin prick test (SPT) and specific Immunglobulin (Ig) E-levels. Isolated tomato seed total protein showed high SPT activity comparable or even higher than tomato fruit protein. For the molecular characterization of tomato seed allergens, a multidimensional protein fractionation strategy and LC-MS/MS was used. Two legumin- and vicilin-proteins were purified and showed strong IgE-reactivity in immunoblots. Individual patient sera exhibited varying IgE-sensitivity against the purified proteins. In silico structural modeling indicates high homology between epitopes of known walnut allergens and the detected IgE-crossreactive tomato proteins.