The biology of the combretastatins as tumour vascular targeting agents.
The tumour vasculature is an attractive target for therapy. Combretastatin A-4 (CA-4) and A-1 (CA-1) are tubulin binding agents, structurally related to colchicine, which induce vascular-mediated tumour necrosis in animal models. CA-1 and CA-4 were isolated from the African bush willow, Combretum caffrum, and several synthetic analogues are also now available, such as the Aventis Pharma compound, AVE8062. More soluble, phosphated, forms of CA-4 (CA-4-P) and CA-1 (CA-1-P) are commonly used for in vitro and in vivo studies. These are cleaved to the natural forms by endogenous phosphatases and are taken up into cells. The lead compound, CA-4-P, is currently in clinical trial as a tumour vascular targeting agent. In animal models, CA-4-P causes a prolonged and extensive shut-down of blood flow in established tumour blood vessels, with much less effect in normal tissues. This paper reviews the current understanding of the mechanism of action of the combretastatins and their therapeutic potential.


Risk factors of polysensitization to contact allergens.
'Polysensitization' (PS) is usually defined as contact sensitization to three or more unrelated haptens of the baseline patch test series. Despite PS being an important clinical phenotype indicating increased susceptibility to contact allergy, statistical approaches to analyse PS have hitherto been preliminary. To apply an appropriate regression model for count data, namely, negative binomial hurdle regression, to a large set of clinical patch test data with the aim of estimating PS risk in more detail than previously achieved. The detailed information provided by the hurdle model includes a separate estimation of an 'increment factor' quantifying the likelihood of further positive reactions, i.e. PS. Clinical data of 126 878 patients patch tested by departments comprising the IVDK (Information Network of Departments of Dermatology) network (www.ivdk.org) between 1995 and 2010 were included. Regarding anatomical sites as exposure (surrogate), the axillae and the feet were found to be strong PS risk factors. Moreover, age was a strong PS risk factor, and less so, female sex. In comparison, atopic eczema and occupational dermatitis were less important risk factors. Single allergens contributed to PS to a varying extent. The data presented point to some, very likely exposure-related, risk factors which need to be considered in future PS research, e.g. addressing the genetic basis for PS.


An integrative view of the role of oxidative stress, mitochondria and insulin in Alzheimer's disease.
The processes underlying the pathogenesis of Alzheimer's disease involve several factors including impaired glucose/energy metabolism, mitochondrial dysfunction, oxidative stress and altered insulin-signaling pathways. This review is mainly devoted to discuss evidence supporting the notion that mitochondrial dysfunction and oxidative stress are interconnected and intimately associated with the development and progression of Alzheimer's disease. Furthermore, the review explores the role of insulin signaling in the pathophysiology of the disease. Indeed, several studies have begun to find links between insulin and mechanisms with clear pathogenic implications for this disorder. Understanding the key mechanisms involved in the etiopathogenesis of Alzheimer's disease may provide opportunities for the design of efficacious preventive and therapeutic strategies.


The first homologous series of self-assembled aryl bromo- and aryl cyanocuprates, -argentates, and -aurates; MLi(2)XAr(2) (M = Cu(I), Ag(I), Au(I); X = Br, C(triple bond)N; Ar = [C(6)H(4)CH(2)N(Et)CH(2)CH(2)NEt(2)-2]-).
Reaction of 2 molar equiv of the diamine chelated aryllithium dimers Li(2)(C(6)H(4)[CH(2)N(Et)CH(2)CH(2)NEt(2)]-2)(2) (Li(2)Ar(2)) with the appropriate metal bromide allows the synthesis of the first homologous series of monomeric group 11 bromoate complexes of type MLi(2)BrAr(2) (M = Cu (7), Ag (8), Au (9)). Both in the solid state and in solution, the bromocuprate 7 is isostructural with the bromoargentate 8. The crystal structures of 7 and 8 consist of a MLi(2) core, and each of the two aryl ligands bridges via electron-deficient bonding between the group 11 metal and one Li atom (d(C(ipso)-M) = 1.941(4) (mean) and 2.122(4) (mean) A, for 7 and 8, respectively). The bromine atom exclusively bridges between the two lithium atoms. Each of the ortho-CH(2)N(Et)CH(2)CH(2)NEt(2) moieties is N,N'-chelate bonded to one lithium (d(N-Li) = 2.195(5) and 2.182(0) (mean) A for 7 and 2.154(8) and 2.220(1) (mean) A for 8). Although the MLi(2)BrAr(2) compounds are neutral higher-order -ate species, the structure can also be regarded as consisting of a contact ion pair consisting of two ionic fragments, [Li-Br-Li](+) and [Ar(2)M](-), which are interconnected by both Li-N,N'-chelate bonding and a highly polar C(ipso)-Li interaction. On the basis of NMR and cryoscopic studies, the structural features of the bromoaurate 9 are similar to those of 7 and 8. A multinuclear NMR investigation shows that the bonding between the [Li-Br-Li] and [Ar(2)M] moieties is intermediate between ionic and neutral with an almost equally polarized C(ipso)-Li bond in 7, 8, and 9. Similar reactions between M(C(triple bond)N) and 2 molar equiv of LiAr yield the analogous 2:1 cyanoate complexes of type MLi(2)(C(triple bond)N)Ar(2) (M = Ag (10), Au (11)). Multinuclear NMR studies show that the cyanoate complexes 10 and 11 are isostructural with the bromoate complexes 7, 8, and 9. This paper illustrates that these cyanoaurates may serve as excellent model complexes to gain more insight into the structure of 2:1 cyanocuprates in solution.


Prenatal Amphetamine-Induced Dopaminergic Alteration in a Gender- and Estrogen-Dependent Manner.
Prenatal exposure to amphetamine induces changes in dopamine receptors in mesolimbic areas and alters locomotor response to amphetamine during adulthood. Sex differences have been reported in amphetamine-induced brain activity and stress sensitivity. We evaluated the effects of prenatal amphetamine exposure on locomotor activity, dopamine receptors and tyrosine hydroxylase mRNA expression in nucleus accumbens and caudate-putamen in response to amphetamine challenge in adult female and male rats. The role of estrogen in the response to restraint stress was analyzed in ovariectomized, prenatally amphetamine-exposed rats. Pregnant rats were treated with D-amphetamine during days 15-21 of gestation. Nucleus accumbens and caudate-putamen were processed for mRNA determination by real-time PCR. In nucleus accumbens, higher mRNA dopamine (D3) receptor expression was found in basal and D-amphetamine-challenge conditions in female than male, and prenatal amphetamine increased the difference. No sex differences were observed in caudate-putamen. Basal saline-treated females showed higher locomotor activity than males. Amphetamine challenge in prenatally amphetamine-exposed rats increased locomotor activity in males and reduced it in females. In nucleus accumbens, estrogen diminished mRNA D1, D2 and D3 receptor expression in basal, and D1 and D3 in ovariectomized stressed rats. Estrogen prevented the increase in tyrosine hydroxylase expression induced by stress in ovariectomized prenatally exposed rats. In conclusion, estrogen modulates mRNA levels of D1, D2 and D3 receptors and tyrosine hydroxylase expression in nucleus accumbens; prenatal amphetamine-exposure effects on D3 receptors and behavioral responses were gender dependent.


Antioxidants in treating pathologies involving oxidative damage: an update on medicinal chemistry and biological activity of stobadine and related pyridoindoles.
Pathologies involving oxidative stress are indicative of malfunction of endogenous antioxidant capacity. Numerous efforts were made to design and synthesize biologically active antioxidants and free oxygen radical scavenging substances that could improve the endogenous antioxidant status. The antioxidant and reactive-oxygen-species-scavenging activity of STB was well demonstrated in many in vitro and in vivo studies. These properties of STB seem to be closely related to its beneficial effects in models of oxidative-stress-involving pathologies, including myocardial infarction, stroke, neurodegenerative disorders, hypoxic-ischemic tissue injury, diabetic complications, chronic inflammation, etc. STB has a good affinity to lipids and exerts its protective activity against free-radical-mediated damage by preventing lipid peroxidation. Rather than interacting with the radicals initiating lipid peroxidation, STB was shown to act in its propagation stage via scavenging peroxyl and/or alkoxyl radicals. STB was also found to protect proteins, predominantly by a mechanism involving protection of thiol groups and by preventing oxidation of amino acids. The first findings on antioxidant and pharmacodynamic effects of STB, tested in a variety of biological models, were summarized in 1998. Recently, chemical modification of STB, which we considered as the leading structure, led to the synthesis of pyridoindole derivatives with significantly increased intrinsic antiradical activity and overall antioxidant efficacy compared to the parental molecule. The present paper provides a complete overview of the literature published since 1998 on both STB and STB congeners. Moreover, appropriate structural modifications of STB provided the opportunity to modulate lipophilicity and acidobasic behavior, thus optimizing bioavailability of the novel derivatives and attenuating their unwanted sideeffects, with the result of decreased toxicity. Hence, STB congeners might be prospectively used as medicinal antioxidants, i.e. remedies effective in conditions involving oxidative stress-mediated injury.


Effects of metformin plus gliclazide compared with metformin alone on circulating endothelial progenitor cell in type 2 diabetic patients.
Circulating endothelial progenitor cells (EPCs) play an important role in the development and progression of diabetic vascular complications. The aim of this study was to investigate the effects of gliclazide plus metformin (GLIMET) compared with metformin alone (MET) on number and function of circulating EPCs in T2DM patients. Patients with newly diagnosed T2DM were randomly divided into two groups, receiving the following treatments for 16 weeks: MET group (assuming metformin 500-2500 mg/day, n=24) and GLIMET group [assuming gliclazide (modified release, 30-60 mg/day)+metformin (250-1000 mg/day), n=23]. Circulating EPCs were quantified by flow cytometry, and the ability to uptake LDL and stain for lectin were used as another method of characterizing EPCs ex vivo. The functions of circulating EPCs were evaluated by colony-forming units (CFU) and migration. The status of oxidative stress was analyzed by serum-free malonaldehyde (MDA) and superoxide dismutase (SOD). There were no significant differences in clinical characteristics and number and function of circulating EPCs between two groups at baseline. Glycemic responses were similar after treatments. Compared with MET group, GLIMET group was associated with an increase in circulating EPCs number, DiLDL-lectin-positive EPCs, and migration. The mean improvements in MDA and SOD of GLIMET group were more strongly upregulated than those of MET group. This study demonstrated that both metformin mono-treatment and metformin plus gliclazide combination treatment provided with improvements in number and function of circulating EPCs. Compared with metformin mono-treatment, early use of combination therapy with gliclazide plus metformin made more effective improvements in circulating EPCs.


Obesity worsens the outcome of influenza virus infection associated with impaired type I interferon induction in mice.
Increasing evidence indicates that obesity is a risk factor for increased severity of influenza virus infection. However, its precise immunological mechanism is not fully understood. To investigate this, diet-induced obese (DIO) mice were established by feeding C57BL/6 male mice a high-fat diet for 16 weeks. DIO and lean control mice were infected intranasally with 3000 pfu of influenza A virus (IAV) (PR8/H1N1). Interestingly, we found adipose tissue located along the bronchus in naïve DIO mice. In addition, the Nos2 level was significantly higher and Arg1 level was significantly lower in lung macrophages of naïve DIO mice, consistent with an M1-skewed phenotype. The survival rate and body weight of DIO mice infected with IAV were significantly lower than those of lean control mice and associated with higher viral load in the lungs of DIO mice. Histopathological analysis demonstrated higher numbers of inflammatory cells in the lungs of DIO mice after IAV infection. Levels of cytokines, including TNF-α, IL-6, IL-10, and type I IFN (IFN-α and IFN-β), in bronchoalveolar lavage fluid (BALF) were altered after IAV infection; in particular, IFN-α and IFN-β levels were significantly suppressed in the BALF of DIO mice. In vitro, bone marrow-derived macrophages were stimulated with ligands of toll-like receptor (TLR) 7/8, a pattern recognition receptor for single-stranded RNA, and levels of TNF-α, IL-6, and IL-10 were similarly altered. In addition, levels of IFN-α and IFN-β were significantly lower in culture supernatants of alveolar macrophages sorted from naïve DIO mice and infected with IAV, compared to those in macrophages sorted from lean control mice. Collectively, these results suggest that macrophages may be the main contributors to poor outcomes of influenza virus infection in obesity.


Portable device for characterizing the angular response of UV spectroradiometers.
This paper introduces a device that was developed to measure the angular response of UV spectroradiometers in the field. This device is designed to be used at the operating position of spectroradiometers; thus the derived angular response also includes any effects from imperfect leveling of the diffuser and corresponds to the actual operational angular response. The design and characterization of the device and the results from its application on 11 different spectroradiometers that operate at different European UV stations are presented. Various sources of uncertainties that were identified result in a combined uncertainty in determining the angular response, which ranges between approximately 1.5% and 10%, depending on the incidence angle and the characteristics of the diffuser. For the 11 instruments, the error in reporting the diffuse irradiance ranges between 2% and - 13%, assuming isotropic distribution of the downwelling radiances.


[Cytometric evaluation of immaturity granulocytes in infections].
The leukocyte differential count is one of the most useful and frequency requested tests in the clinical laboratory. Conventional blood cell counting instruments can enumerate only the WBCs normally found in the peripheral blood, i.e., neutrophils, lymphocytes, monocytes, eosinophils, and basophils. Nevertheless, there are other cells not normally found in the peripheral blood, such as immature granulocyte. They are indicators of different disease states. We haven't got still references of automatic method to good evaluation of enumeration of the immature granulocyte. The aim of this study was compare count immature granulocytes by manual microscopy method with flow cytometric method. Flow cytometric method enumeration of immature granulocyte was performed using monoclonal antibody: anti-CD45, anti-CD11b and anti-CD16. A comparison of flow cytometry count with the manual count of promyelocytes, myelocytes, and metamyelocytes using linear regression revealed a correlation coefficient of 0,85 for percentage counts. Our results indicate that the flow cytometric immature granulocyte count can replace the manual morphology count for the laboratory work.


[THE CELLULAR MECHANISMS OF LUNG EDEMA CLEARANCE: DOES THE ALVEOLAR EPITHELIUM PLAY A ROLE?]
Pulmonary edema develops as a result of either alteration in the hydrostatic and oncotic pressure gradients across the pulmonary circulation and the lung interstitium or due to increased lung permeability. Alveolar fluid clearance is important in keeping the airspaces free of edema. This process is carried out via the alveolar epithelial active transport of Na+ across the alveolo-capillary barrier mostly by apical Na+ channels and basolateral Na,K-ATPases. Several pharmacologic agents such as catecholamines, vasopressin and gene therapy interventions have currently been found to stimulate the active Na+ transport and lung edema clearance. While others such as amiloride, ouabain, high tidal volume ventilation, hyperoxia and sepsis decrease the rate of alveolar fluid clearance. In conclusion, this review discusses the mechanisms and signal pathways by which the alveolar epithelium impacts lung edema clearance.


The cell of origin of ovarian epithelial tumors and the ovarian surface epithelium dogma: does the emperor have no clothes?
The widely favored hypothesis that ovarian epithelial tumors arise from the mesothelial cell layer lining the ovarian surface fails to explain the resemblance of these tumors to those arising in organs that are embryologically derived from the Müllerian ducts such as fallopian tubes, endometrium, and endocervix. In addition, this theory cannot account for the fact that tumors that are morphologically identical to ovarian carcinomas can sometimes be found outside the ovary. A suggestion is made that components of the secondary Müllerian system, which include paraovarian/paratubal cysts, rete ovarii, endosalpingiosis, endometriosis, and endomucinosis, merit some consideration as to their possible role in ovarian tumorigenesis.


Effect of Nell-1 delivery on chondrocyte proliferation and cartilaginous extracellular matrix deposition.
Cartilage tissue engineering using chondrogenic growth factors is an attractive strategy to promote cartilage repair. Bone morphogenetic proteins have been widely studied for their application in cartilage repair. However, functional heterogeneity of bone morphogenetic proteins and unpredictable effects such as cyst formation may limit their therapeutic use. Thus, the use of alternative growth factors with greater osteochondral specificity may be advantageous for cartilage regeneration. Nel-like molecule-1 (Nell-1; Nel is a protein strongly expressed in neural tissue encoding epidermal growth factor-like domain) is a novel growth factor believed to specifically target cells committed to the osteochondral lineage. Mutation of the Nell-1 gene has been shown to disrupt normal cartilage growth and development in rodents. This study investigates the chondrogenic potential of recombinant human Nell-1 protein in a three-dimensional alginate hydrogel microenvironment containing rabbit chondrocytes. To provide controlled delivery and maximize biological efficiency, Nell-1 was incorporated in chitosan microparticles. Over 42 days of culture, chondrocyte proliferation and cluster formation was significantly enhanced by Nell-1 in a dose-dependent manner. Further, the clusters formed in the presence of Nell-1 contained more type II collagen and glycosaminoglycans than clusters formed within Nell-free control gels. These findings demonstrate the ability of Nell-1 to promote chondrocyte proliferation and deposition of cartilage-specific extracellular matrix materials.


[Clinical and physiopathologic study of the hepatotoxicity of psychotropic drugs including the antiepileptics].
The goal of this article is to inform psychiatrists of the hepatotoxicity of certain drugs used for the treatment of psychiatric disorders. A review of biochemical pathways lends support to the argument that toxicity is due to certain toxic metabolites. The physiopathology of these disorders, which are rare, requires an appreciation of an individual's biochemical make up which can explain the quantity of active metabolites produced, the occurrence of immunological reactions and the contributing factor of enzymatic induction resulting from the simultaneous prescription of several medications. The most serious disorder is cytotoxic hepatitis which has a 10 to 20 per cent mortality. The molecules which may cause this affection in less than one out of a thousand cases are the hydrazide M.A.O. inhibitors and valproic acid. These medications should not be prescribed without being aware of their drawbacks and after a failure to achieve improvement with other drugs having a similar action. Obviously it is important to prescribe no more than moderate doses, and to avoid prescribing medications which, together can occasion enzymatic induction. The most common complication is obstructive jaundice for which the clinical and biological diagnosis should be made early in the course of the disorder. The medication should be stopped immediately and further prescription, in patients who have already presented this disorder, when a molecule is known to have certain toxic side effects (phenothiazines and tricyclics), suspended this surveillance should be carried out over a three months period. The occurrence of jaundice during a period of treatment is not always iatrogenic. It is always necessary to ascertain the etiology by asking a specialist to do a complete workup.


[Endophthalmitis following cataract surgery].
Infectious endophthalmitis is a serious complication following cataract surgery, since it often induces a substantial reduction of visual acuity. We retrospectively evaluated the clinical data of 53 patients with endophthalmitis following cataract surgery who were treated at the department of ophthalmology of the University Hospital in Ulm between 1995 and May 2001. Of these patients, 50 had been referred. Clinical presentation, infecting organism, treatment and visual outcome were analysed with a followup ranging from 2 weeks up to 42 months (median: 6 months). In 52 patients endophthalmitis was preceeded by cataract extraction and IOL implantation, in one case by secondary IOL implantation. Confirmed microbiologic growth was demonstrated from intraocular specimens in 26 out of 41 operated eyes (63%), the most frequent causative organisms were coagulase-negative Staphylococci (50%). All isolated bacteria were sensitive to a combination of the antibiotics vancomycin and amikacin or vancomycin and ceftazidime. 13 patients were treated with intravenous antibiotic therapy alone. In 46% of patients, who were initially treated with intraocular antibiotic injections alone, required further therapeutic intervention for recurrent infection. Only 7.7% of the patients who initially underwent intraocular antibiotic injections combined with IOL removal or pars plana vitrectomy with or without IOL removal, required further surgical intervention. Initial visual acuity was hand movements (median) only but improved during follow-up to 0.2 (median). In this series all tested bacteria were susceptible to the combination of vancomycin with either amikacin or ceftazidime. Aggressive initial treatment including IOL removal may be associated with a lower frequency of recurrent disease.


Proliferative behavior of the murine cerebral wall in tissue culture: cell cycle kinetics and checkpoints.
Cerebral wall from embryonic day 13 mice was cultured in a three-dimensional collagen matrix in defined, serum-free medium. The cerebral wall retained its normal architecture, including the radial glial fiber system, for up to 19 h in culture. The cell cycle was initially blocked at the S/G2/M and the G1/S phase transitions, resulting in a transient synchronization of the proliferative cells. The transient blockades correspond, we suggest, to the G2 checkpoint and G1 restriction point, adaptive mechanisms of normal proliferative cells. The blocks were relieved within a few hours of explantation with restoration of the interkinetic nuclear migration and flow of cells through the cycle phases. The duration of the reestablished cell cycle and those of G1, S, and combined G2-M phases were estimated to be 19.2, 6.3-8.3, 8.8, and 2.0-4.0 h, respectively. The leaving (Q) fraction of the cycle (0.64) was twice the in vivo value. Two-thirds of the Q fraction cells remained in the ventricular epithelium, resulting in a substantially low growth fraction of 0.73 compared with 1.0 in vivo. The embryonic murine cerebral explant, cultured in minimum essential medium, should be favorable for studies of cycle modulatory actions of cell external influences such as growth factors or neurotransmitters.


Novel glucagon receptor antagonists with improved selectivity over the glucose-dependent insulinotropic polypeptide receptor.
Optimization of a new series of small molecule human glucagon receptor (hGluR) antagonists is described. In the process of optimizing glucagon receptor antagonists, we counter-screened against the closely related human gastric inhibitory polypeptide receptor (hGIPR), and through structure activity analysis, we obtained compounds with low nanomolar affinities toward the hGluR, which were selective against the hGIPR and the human glucagon-like peptide-1 receptor (hGLP-1R). In the best cases, we obtained a >50 fold selectivity for the hGluR over the hGIPR and a >1000 fold selectivity over the hGLP-1R. A potent and selective glucagon receptor antagonist was demonstrated to inhibit glucagon-induced glycogenolysis in primary rat hepatocytes as well as to lower glucagon-induced hyperglycemia in Sprague-Dawley rats. Furthermore, the compound was shown to lower blood glucose in the ob/ob mouse after oral dosing.


Acute pericarditis caused by Streptococcus pneumoniae in young infants and children: three case reports and a literature review.
We present three cases of pericarditis caused by Streptococcus pneumoniae diagnosed in children. The presenting symptoms were fever, dyspnea, tachycardia, and hepatomegaly. Chest X-rays showed an enlarged heart silhouette and echocardiographic examination revealed a pericardial effusion in all cases. S. pneumoniae was recovered from the pericardial fluid and/or blood in all cases. Systemic antibiotic treatment and dopaminergic agents were used; all patients recovered completely. Another 10 cases of S. pneumoniae pericarditis in children reported since 1980 are presented. Most cases had preceding respiratory symptoms and had a good outcome. While rare and life-threatening, pericarditis caused by S. pneumoniae has a good outcome if diagnosed early and treated adequately.


P-fimbriae studies on the diagnosis and prevention of acute pyelonephritis.
Theoretically there are several ways to prevent pyelonephritis and renal scarring caused by P-fimbriated Escherichia coli. Screening for individuals at risk, e.g. those carrying P-fimbriated pyelonephritogenic E. coli or those with high receptor density on their uroepithelial cells, could perhaps define a population where prophylaxis with a receptor analogue or vaccination with P-fimbriae may be relevant. Epidemiological measures in neonatal and maternity wards may prevent the nosocomial spread of virulent bacteria and reduce the number of colonized infants. However, no such methods have so far had any proven clinical relevance, and today, the important concern is still to try by conventional means--as we have always done--to get an early diagnosis and to treat the patient without delay.


Monte Carlo Simulation of Radiotherapy for Breast Cancer in Pregnant Patients: How to Reduce the Radiation Dose and Risks to Fetus?
This study estimated the fetal dose and risks from radiotherapy for breast cancer with 6 MV X-rays. Breast irradiation was simulated with the MCNP code using two mathematical phantoms corresponding to patients in the early and middle periods of pregnancy. Monte Carlo simulations were performed to determine the appropriate fetal shielding. For a 50-Gy tumor dose, the unshielded fetal dose reached up to 133.1 mGy. Fetal protection with a lead shield of dimensions 30 × 30 × 5 cm3 placed besides the treatment couch resulted in maximum doses of 22.0 and 70.3 mGy at the first and second trimesters of gestation, respectively. These shielded fetal doses may be associated with a fatal cancer risk during childhood up to 0.42% and a maximum probability for the appearance of heritable effects of 0.17%. The use of fetal shielding ensures the absence of deterministic effects from radiotherapy during the first 24 weeks of gestation.


Discovery and optimization of thienopyridine derivatives as novel urea transporter inhibitors.
Urea transporters (UTs) play an important role in the urine concentrating mechanism and are recognized as novel targets for developing small molecule inhibitors with salt-sparing diuretic activity. Thienoquinoline derivatives, a class of novel UT-B inhibitors identified by our group, play a significant diuresis in animal model. However, the poor solubility and low bioavailability limited its further development. To overcome these shortcomings, the structure modification of thienoquinoline was carried out in this study, which led to the discovery of novel thienopyridine derivatives as specific urea transporter inhibitors. Further optimization obtained the promising preclinical candidate 8n with not only excellent inhibition effect on urea transporters and diuretic activity on rat model, but also suitable water solubility and Log P value.


Non-symmetrical cytosine methylation in tobacco pollen DNA.
We have detected sequence-specific non-symmetrical cytosine methylation within a 140 bp region of the promoter for the tobacco auxin-binding protein gene T85 in pollen DNA. Direct sequencing of the population of bisulphite reaction products showed that, in this region. 10 out of a possible 49 cytosine residues were methylated at a high frequency in pollen whereas the corresponding region from somatic cells (leaf DNA) did not show a detectable level of methylation. The context of these sites was 1 x m5CpTpC, 1 x m5CpGpT, 1 x m5CpCpT, 2 x m5CpTpT, 2 x m5CpGpG, and 3 x m5CpApT of which only m5CpGpG and m5CpGpT fitted the consensus sequence for symmetrical methylation in plants.


Analgesic, anticonvulsant and antioxidant activities of 3-[4-(3-trifluoromethyl-phenyl)-piperazin-1-yl]-dihydrofuran-2-one dihydrochloride in mice.
Recently we have shown that 3-[4-(3-trifluoromethyl-phenyl)-piperazin-1-yl]-dihydrofuran-2-one dihydrochloride (LPP1) is an antinociceptive and local anesthetic agent in rodents. Below an extended study of the pharmacological activity of LPP1 is described. In vitro LPP1 has no affinity for GABA(A), opioidergic μ and serotonergic 5-HT(1A) receptors. The total antioxidant capacity of LPP1 (1-10mM) measured as ABTS radical cation-scavenging activity showed that LPP1 has dose-dependent antioxidant properties in vitro. Low plasma concentration of this compound detected by means of HPLC method 30min after its intraperitoneal administration suggests a rapid conversion to metabolite(s) which may be responsible for its analgesic and anticonvulsant activities in vivo. In vivo the compound's influence on the electroconvulsive threshold and its activity in the maximal electroshock seizure test (MES) were evaluated. The results demonstrated that LPP1 had an anticonvulsant activity in the MES model (ED(50)=112mg/kg) and at a dose of 50mg/kg was able to elevate the electroconvulsive threshold for 8mA as compared to the vehicle-treated mice. The analgesic activity of LPP1 was investigated in the acetic acid-induced writhing test in two groups of mice: animals with sensory C-fibers ablated, and mice with C-fibers unimpaired. It proved the potent activity of this compound in both groups (approximately 85% as compared to the vehicle-treated mice). The adverse effects of LPP1 were evaluated as acute toxicity (LD(50)=747.8mg/kg) and motor coordination impairments in the rotarod and chimney tests. The results from these tests show that LPP1 at doses higher than 100mg/kg is likely to impair the motor performance of experimental animals. Concluding, LPP1 is an analgesic and anticonvulsant compound which has antioxidant properties in vitro. Further studies are necessary to assess whether the antioxidant activity and the receptor profiling demonstrated in vitro can be confirmed for its metabolite(s) that are formed in vivo.


Chemical sterilization with an IUD.
Zipper has more than adequately demonstrated that quinacrine, when placed in the uterine cavity, has a sclerosing effect on the tubal ostia, resulting in chemical sterilization of the fallopian tubes. His technique involves the use of 3 monthly instillations of 250 mg of pellets into the uterine cavity. With the hope of reducing the total dosage and making it a 1-insertion technique, quinacrine has been mixed with polyethylene oxide and place on the arms of various IUDs. It is hoped that the material is directed at the tubal ostia. Studies to date have occurred in women awaiting hysterectomy with careful pathologic examination of the intramural portion of the tube. The results thus far are most encouraging and once the best vector is identified, it is hoped that this will become an acceptable means of female sterilization.


Identification of circRNAs involved in the development of hepatocellular carcinoma after insufficient radiofrequency ablation.
Previous studies have reported that circular RNAs (circRNAs) play a key role in the pathogenesis and progression of various diseases. In the present study, we aimed to identify potential circRNAs associated with the progression of hepatocellular carcinoma (HCC) after insufficient radiofrequency ablation (IRFA). A xenograft mouse IRFA model was initially established, and immunohistochemical staining (IHC) and polymerase chain reaction (PCR) were performed to confirm the expression of programmed cell death-ligand 1 (PD-L1) and vascular endothelial growth factor receptor-1 (VEGFR-1). CircRNA expression alterations were screened by next-generation sequencing (RNA-seq). Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were conducted to predict the function of genes coding differentially expressed circRNAs. The selected circRNAs were validated utilizing PCR and Sanger sequencing. The relationships between circRNAs, microRNAs, PD-L1, and VEGFR-1 were predicted by bioinformatics. Overall, a total of 612 circRNAs were differentially expressed in IRFA-treated subcutaneous tumorigenesis tissue. Among them, 435 circRNAs were significantly upregulated and 177 circRNAs were downregulated. GO and KEGG analyses were employed to predict the functions of these circRNAs. Thereafter, quantitative reverse transcription PCR (qRT-PCR) assays determined that these seven circRNAs were overexpressed in the IRFA group, which was consistent with the RNA-seq results. Based on the bioinformatic analysis, seven circRNAs confirmed by Sanger sequencing were predicted to likely regulate PD-L1 and VEGFR-1 expression levels by functioning as sponges for microRNAs (miRNAs) and forming a circRNA-miRNA-PD-L1/VEGFR-1 regulatory network. Finally, IHC and qRT-PCR of PD-L1 and VEGFR-1 confirmed the activation of this pathway. Taken together, we report that differentially expressed circRNAs might simultaneously regulate PD-L1 and VEGFR-1 in the IRFA tissues, which provides a novel view of circRNAs in HCC progression after the IRFA procedure.


A novel role of Id-1 in regulation of epithelial-to-mesenchymal transition in bladder cancer.
Inhibitor of differentiation or DNA binding -1 (Id-1) has been shown to be increased in several types of advanced cancer, and to be associated with aggressive and metastatic abilities of cancer cells. Recently, more and more evidence indicates that epithelial-to-mesenchymal transition (EMT) is an important mechanism taking place during tumor invasion and metastasis, but the molecular pathways underlying EMT have not been clearly established. This study was to investigate the expression of Id-1 in bladder cancer and its association with EMT. A total of 169 tissues, consisting of 147 primary bladder cancers and 22 adjacent normal tissues were included in this study. Id-1, E-cadherin, and β-catenin were examined immunohistochemically in paraffin sections. The pBabe-Id-1 expression retroviral vector and retroviral vectors containing an Id-1-specific small interfering RNA oligonucleotides (si-Id-1) were transfected into 2 bladder cancer cell lines respectively. Then, we used Western blotting and immunofluorescent staining to detect the cellular expression of epithelial markers and mesenchymal markers. The invasion and migration ability of bladder cancer cells were identified by type I collagen invasion assay and wound closure assay. We demonstrated that increased Id-1 expression was associated with advanced tumor stage and grade. In addition, the increased Id-1 expression in bladder tumors was also correlated with decreased membranous E-cadherin and β-catenin expression. In vitro, studies showed that inactivation of the Id-1 gene conferred morphologic transition of bladder cancer cells from a fibroblastic to epithelial appearance, and overexpression of Id-1 could lead to acquisition of a fibroblastic spindle cell phenotype accompanied by loss of cell-to-cell contacts. By Western blotting and immunofluorescent staining, we showed that the expression level of Id-1 was correlated with the expression of mesenchymal markers but was inversely correlated with the expression of epithelial markers. Moreover, results of collagen invasion and wound closure assays showed ectopic Id-1 expression led to increased ability of invasion and migration. Our results suggest that Id-1 may play roles in tumor progression and EMT activation in bladder cancer.


Mandatory overtime regulations and nurse overtime.
A descriptive study used data from the 2004 National Sample Survey of Registered Nurses to examine the nature and occurrence of RN mandatory, voluntary overtime, paid on-call, and total work hours and their association with mandatory overtime regulations in United States. About half of the nurses worked more than 40 hrs per week. Nurses working in states regulating mandatory overtime reported lower levels of mandatory overtime hours than states without regulations or states restricting total work hours. The percent of RNs working 61 hrs and over per week in states without regulations was lower than that in states with regulations. Nurses working in nursing homes reported higher levels of the percentage of mandatory overtime hours worked than those working in hospitals. This suggested that governments need to continuously supervise healthcare institutions, including both hospitals and nursing homes, to ensure adherence to mandatory overtime regulations.


Development and perspectives of rapid detection technology in food and environment.
Food safety become a hot issue currently with globalization of food trade and food supply chains. Chemical pollution, microbial contamination and adulteration in food have attracted more attention worldwide. Contamination with antibiotics, estrogens and heavy metals in water environment and soil environment have also turn into an enormous threat to food safety. Traditional small-scale, long-term detection technologies have been unable to meet the current needs. In the monitoring process, rapid, convenient, accurate analysis and detection technologies have become the future development trend. We critically synthesizing the current knowledge of various rapid detection technology, and briefly touched upon the problem which still exist in research process. The review showed that the application of novel materials promotes the development of rapid detection technology, high-throughput and portability would be popular study directions in the future. Of course, the ultimate aim of the research is how to industrialization these technologies and apply to the market.


Effects of teriparatide, alendronate, or both on bone turnover in osteoporotic men.
We have previously demonstrated that alendronate reduces the ability of teriparatide to increase bone mineral density (BMD) in osteoporotic men. The underlying basis for this observation is poorly understood. The primary aim of this study was to determine whether teriparatide increases osteoblast activity when the ability of teriparatide to increase osteoclast activity is suppressed by alendronate. This was a nonblinded, randomized, controlled trial. The study was conducted at the General Clinical Research Center of a teaching hospital. We studied 63 men, age 46-85, with low spine and/or hip BMD. Subjects received alendronate 10 mg daily (group 1), teriparatide 37 microg sc daily (group 2), or both (group 3) for 30 months. Teriparatide was begun at month 6. The primary endpoint was the change in serum N-telopeptide, osteocalcin, and amino-terminal propeptide of type 1 procollagen. In men receiving teriparatide monotherapy (group 2), levels of all bone turnover markers increased markedly during the first 6 months of teriparatide administration and then declined toward baseline during the next 18 months. In men who received combination therapy (group 3), bone turnover marker levels declined in the first 6 months (while receiving alendronate alone) and then returned to baseline levels (N-telopeptide) or above (osteocalcin and amino-terminal propeptide of type 1 procollagen) after teriparatide was added. Changes in each marker were significantly different between groups 1 and 2 (all P values < 0.001), groups 1 and 3 (all P values < 0.001), and groups 2 and 3 (all P values < 0.03). As with BMD, alendronate impairs the action of teriparatide to increase bone turnover in men.


Effect of Nano-Y2O3 on the Microstructure and Properties of WC-Reinforced Ni-Based Composite Surfacing Layer.
In this study, a WC-reinforced Ni-based surfacing layer was prepared on Q235 steel plate by plasma arc welding. The effects of nano-Y2O3 with different contents (0 wt.%, 0.4 wt.%, 0.8 wt.%, 1.2 wt.%, and 1.6 wt.%) on the microstructure, phase composition, microhardness, and wear resistance of the surfacing layer were studied by scanning electron microscope (SEM), energy dispersive spectrometer (EDS), X-ray diffraction (XRD), microhardness test, and pin-on-disk test. The results show that the phase composition of the surfacing layer was γ-Ni, FeNi3 solid solution, WC, W2C, M23C6, M6C, Cr7C3, and other carbides. When the addition of nano-Y2O3 was 1.2 wt.%, it has a good improvement on microstructure grain refinement and carbide hard-phase increase. Compared with other contents, 1.2 wt.% nano-Y2O3 surfacing layer has the highest microhardness and the lowest friction coefficient and wear loss. At this time, the wear mechanism is abrasive wear accompanied by slight adhesive wear.


Drug treatment for HIV-1-associated dementia.
The manifestation of HIV-1-associated minor motor deficits or HIV-1-associated dementia indicates viral activity within the CNS. Highly active antiretroviral therapy (HAART) has a prophylactic effect, which does not protect all patients, because different pathomechanisms are involved. A CNS-specific therapeutic efficacy can be demonstrated for nucleoside and non-nucleoside reverse transcriptase inhibitors (NRTIs; NNRTIs). To be effective in the CNS, HAART should contain either zidovudine or stavudine and NNRTI should be added if there is dinical or preclinical evidence of CNS involvement. Protease inhibitors may be sufficient to lower high systemic viral burden and thus protect the CNS from HIV-1 invasion.


Pioglitazone treatment increases COX-2-derived prostacyclin production and reduces oxidative stress in hypertensive rats: role in vascular function.
PPARγ agonists, glitazones, have cardioprotective and anti-inflammatory actions associated with gene transcription interference. In this study, we determined whether chronic treatment of adult spontaneously hypertensive rats (SHR) with pioglitazone alters BP and vascular structure and function, and the possible mechanisms involved. Mesenteric resistance arteries from untreated or pioglitazone-treated (2.5 mg·kg⁻¹ ·day⁻¹ , 28 days) SHR and normotensive [Wistar Kyoto (WKY)] rats were used. Vascular structure was studied by pressure myography, vascular function by wire myography, protein expression by Western blot and immunohistochemistry, mRNA levels by RT-PCR, prostanoid levels by commercial kits and reactive oxygen species (ROS) production by dihydroethidium-emitted fluorescence. In SHR, pioglitazone did not modify either BP or vascular structural and mechanical alterations or phenylephrine-induced contraction, but it increased vascular COX-2 levels, prostacyclin (PGI₂) production and the inhibitory effects of NS 398, SQ 29,548 and tranylcypromine on phenylephrine responses. The contractile phase of the iloprost response, which was reduced by SQ 29,548, was greater in pioglitazone-treated and pioglitazone-untreated SHR than WKY. In addition, pioglitazone abolished the increased vascular ROS production, NOX-1 levels and the inhibitory effect of apocynin and allopurinol on phenylephrine contraction, whereas it did not modify eNOS expression but restored the potentiating effect of N-nitro-L-arginine methyl ester on phenylephrine responses. Although pioglitazone did not reduce BP in SHR, it increased COX-2-derived PGI₂ production, reduced oxidative stress, and increased NO bioavailability, which are all involved in vasoconstrictor responses in resistance arteries. These effects would contribute to the cardioprotective effect of glitazones reported in several pathologies.


The identification and treatment of opiate users in police custody.
All detainees admitted to seven London police stations were observed over a six-month period (n=2,947). Four per cent were identified as opiate users, although the actual percentage is likely to be much higher. Compared to the general population of detainees there were significantly more women among known opiate users and this group also contained a higher percentage of white detainees and people born in the British Isles. People born in continental European countries were also over-represented. A little more than half of known users did not reveal their use on arrival at the police station. At least 60% of known opiate users remained well throughout their detention, 30% were intoxicated through drugs at the time of their arrest, but only 13% displayed signs or symptoms of withdrawal during their detention. Overall, 65% of the known opiate users were seen by a police surgeon and of these 52% were given medication. All of those withdrawing were given drug treatment, but most of those who were intoxicated by opiates, or who remained well throughout their detention, received no medication. Of those given medication 86% received an opiate, dihydrocodeine being the commonest preparation, usually in association with a benzodiazepine. Despite the adoption of differing management paradigms among police surgeons, the actual medical treatment of opiate-using detainees was pragmatic and determined by individual need.


Curcumin suppresses gastric NF-kappaB activation and macromolecular leakage in Helicobacter pylori-infected rats.
To investigate whether curcumin could attenuate nuclear factor (NF)-kappaB p65 expression and macromolecular leakage in the gastric mucosa of Helicobacter pylori (H. pylori)-infected rats. Twenty-five male Sprague-Dawley rats were equally divided into five groups: control rats (Control), control rats supplemented with 600 mg/kg curcumin, H. pylori-infected rats (Hp), H. pylori-infected rats supplemented with 200 mg/kg curcumin (Hp + curI), and H. pylori-infected rats supplemented with 600 mg/kg curcumin (Hp + curII). In H. pylori-infected groups, rats were inoculated with H. pylori suspension twice a day at an interval of 4 h for 3 d. Two weeks later, 200 or 600 mg/kg curcumin was given once daily to curcumin-supplemented groups for 7 d. On the day of the experiment, macromolecular leakage in gastric mucosa was examined by intravital fluorescence microscopy. The stomach tissue was removed to examine NF-kappaB p65 expression in gastric epithelial cells by immunohistochemistry. The expression of NF-kappaB p65 in gastric epithelial cells and the macromolecular leakage from gastric mucosal microcirculation significantly increased in the Hp group compared with the Control group. The percentages of NF-kappaB p65 immunoreactive cells in Control and Hp groups were 10.72% +/- 2.10% vs 16.02% +/- 2.98%, P = 0.004, respectively. The percentages of macromolecular leakage in Control and Hp groups were 10.69% +/- 1.43% vs 15.41% +/- 2.83%, P = 0.001, respectively. Curcumin supplementation in Hp + curI and Hp + curII groups significantly decreased NF-kappaB p65 immunoreactive cells and macromolecular leakage compared with results in the Hp group. The percentages of NF-kappaB p65 immunoreactive cells in Hp + curI and Hp + curII groups were 11.79% +/- 2.13% (P = 0.017) and 11.42% +/- 1.68% (P = 0.010), respectively. The percentages of macromolecular leakage in Hp + curI and Hp + curII groups were 12.32% +/- 2.13% (P = 0.025) and 12.14% +/- 1.86% (P = 0.018), respectively. H. pylori-induced gastric inflammation in rats is associated with increased NF-kappaB activation and macromolecular leakage which can be reduced by curcumin supplementation.


Cadmium as a tool for studying calcium-dependent cation permeability of the human red blood cell membrane.
Three Ca(2+)-dependent procedures known to increase cation permeability of red blood cell membranes were tested with Cd2+ ions which equal Ca2+ ions both in their charge and the crystal radius, 1. Increase of non-selective permeability for monovalent cations by incubating the red cells in a Ca(2+)-free sucrose medium. Addition of Cd2+ to the suspension of leaky cells failed to restore the initial impermeability of the red cell membrane while a repairing effect of Ca2+ was evident both in the presence and absence of Cd2+. Thus, in low electrolyte medium, Cd2+ could neither mimic Ca2+, nor prevent the latter from interacting with membrane structures which control cation permeability. 2. Increase of the K(+)-selective permeability by propranolol plus Ca2+. Cd2+ added to a Ca(2+)-free Ringer type medium containing propranolol enhanced K+ permeability similar to that obtained with Ca2+. No changes of membrane permeability could be detected in the presence of 0.5 mmol/l Cd2+ in absence of propranolol. The Cd(2+)-stimulated K+ channels were different from those induced by Ca2+. They proved to be insensitive to quinine, exhibited a low K+/Na+ selectivity, and showed no tendency to self-inactivation. 3. Stimulation of K+ permeability by electron donors plus Ca2+. Substitution of Ca2+ by Cd2+ yielded results similar to those obtained with propranolol. The ability of Cd2+ to overtake the role of Ca2+ appears to depend on the system studied. It supplies information allowing to distinguish between the diverse Ca(2+)-dependent systems in cell membranes.


A clinical study of canine collagen type III glomerulopathy.
Collagen type III glomerulopathy (Col3GP), also known as collagenofibrotic glomerulonephropathy, is a rare renal disease with unknown pathogenesis that occurs in animals and humans. We recently described a naturally occurring canine autosomal recessive model of Col3GP, and the aim of the present work was to study the clinical features of canine Col3GP and compare with the human phenotype. In humans two different clinical syndromes with different age at onset (child- or adulthood) have been observed. In children a more aggressive course with familial occurrence is described, characterized by progressively increasing proteinuria, nephrotic syndrome, hypertension and chronic renal failure. A markedly increased serum level of the aminoterminal propeptide of type III procollagen (PIIINP) is considered a useful marker for the disease. Since Col3GP and concurrent hypocomplementemia have been observed in humans, we also aimed to investigate if hypocomplementemia was present in Col3GP affected dogs. A litter consisting of seven puppies, four Col3GP affected and three healthy unaffected, was observed from the day of birth until the affected puppies developed a mild or moderate renal azotemia. During the period of observation growth retardation, increasing blood pressure, progressive proteinuria, azotemia, hypoalbuminemia, hypercholesterolemia and increased serum PIIINP were observed in all the affected dogs. Hypocomplementemia was not detected. Affected dogs were euthanized between 109 and 144 days of age, and pathological examinations revealed ascites and massive glomerular accumulations of collagen type III, consistent with Col3GP. Dogs with Col3GP develop juvenile chronic renal failure, preceded by nephrotic syndrome, elevated serum PIIINP and hypertension, thus have similar clinical features as the juvenile Col3GP in humans. Further studies of this naturally occurring canine phenotype may provide more information on the pathogenesis and genetics of Col3GP in both animals and humans.


Engineered l-Lactate Responding Promoter System Operating in Glucose-Rich and Anoxic Environments.
Bacteria equipped with genetically encoded lactate biosensors are promising tools for biopharmaceutical production, diagnostics, and cellular therapies. However, many applications involve glucose-rich and anoxic environments, in which current whole-cell lactate biosensors show low performance. Here we engineer an optimized, synthetic lactate biosensor system by repurposing the natural LldPRD promoter regulated by the LldR transcriptional regulator. We removed glucose catabolite and anoxic repression by designing a hybrid promoter, containing LldR operators and tuned both regulator and reporter gene expressions to optimize biosensor signal-to-noise ratio. The resulting lactate biosensor, termed ALPaGA (A Lactate Promoter Operating in Glucose and Anoxia), can operate in glucose-rich, aerobic and anoxic conditions. We show that ALPaGA works reliably in the probiotic chassisEscherichia coliNissle 1917 and can detect endogenous l-lactate produced by 3D tumor spheroids with an improved dynamic range. In the future, the ALPaGA system could be used to monitor bioproduction processes and improve the specificity of engineered bacterial cancer therapies by restricting their activity to the lactate-rich microenvironment of solid tumors.


Spiritual Health in Nursing From the Viewpoint of Islam.
In order to gain a more detailed insight into the concept of spiritual health, a hybrid model of concept analysis was used to remove some of the ambiguity surrounding the conceptual meaning of spiritual health in Islamic and Iranian contexts. The purpose of this study was to clarify the meaning and nature of the spiritual health concept in the context of the practice of Islam among Iranian patients. The current concept analysis was undertaken according to the modified traditional hybrid model, which consists of five phases: theoretical phase, initial fieldwork phase, initial analytical phase, and final fieldwork and final analytical phase. In the theoretical phases of the study, the concept of spiritual health was described based on a literature review of publications dealing with the Islamic viewpoint (years: from 2013 to 2014, Databases and search engines: Pubmed, SID, Magiran, Noormax, Google Scholar, Google and IranMex, Languages: English and Persian, Keywords: spiritual health AND (Islam OR Quran), spirituality AND (Islam OR Quran), complete human AND Islam, healthy heart (Galb Salim) AND Islam, healthy life (Hayat tayebeh) AND Islam, calm soul (Nafse motmaeneh) And Islam and healthy wisdom (Aghle Salim) AND Islam). Purposive sampling was conducted and nine participants were selected. Semi-structured interviews and observations were conducted periodically for data collection after obtaining informed consent. Observational, theoretical, and methodological notes were made. Then, using MAXQUDA 7 software, the data were analyzed using qualitative content analysis. The relevant literature in the theoretical phase uncovered the attributes of the concept of spiritual health, including love of the Creator, duty-based life, religious rationality, psychological balance, and attention to afterlife. These attributes were explored in depth in later stages. Finally, the definition of spiritual health was developed. Islam has a unique perspective on spiritual health as it encompasses all aspects of human beings. Thus, it is necessary to carefully study the difference between the Islamic concept of spiritual health and that of other religions and ideologies to design suitable and useful nursing care for Iranian patients that satisfy their spiritual needs.


Increasing physical activity in patients with asthma through positive affect and self-affirmation: a randomized trial.
Patients with asthma engage in less physical activity than peers without asthma. Protocols are needed to prudently increase physical activity in asthma patients. We evaluated whether an educational intervention enhanced with positive-affect induction and self-affirmation was more effective than the educational protocol alone in increasing physical activity in asthma patients. We conducted a randomized trial in New York City from September 28, 2004, through July 5, 2007; of 258 asthma patients, 252 completed the trial. At enrollment, control subjects completed a survey measuring energy expenditure, made a contract to increase physical activity, received a pedometer and an asthma workbook, and then underwent bimonthly follow-up telephone calls. Intervention patients received this protocol plus small gifts and instructions in fostering positive affect and self-affirmation. The main outcome was the within-patient change in energy expenditure in kilocalories per week from enrollment to 12 months with an intent-to-treat analysis. Mean (SD) energy expenditure at enrollment was 1767 (1686) kcal/wk among controls and 1860 (1633) kcal/wk among intervention patients (P = .65) and increased by 415 (95% CI, 76-754; P = .02) and 398 (95% CI, 145-652; P = .002) kcal/wk, respectively, with no difference between groups (P = .94). For both groups, energy expenditure was sustained through 12 months. No adverse events were attributed to the trial. In multivariate analysis, increased energy expenditure was associated with less social support, decreased depressive symptoms, more follow-up calls, use of the pedometer, fulfillment of the contract, and the intervention among patients who required urgent asthma care (all P < .10, 2-sided test). A multiple-component protocol was effective in increasing physical activity in asthma patients, but an intervention to increase positive affect and self-affirmation was not effective within this protocol. The intervention may have had some benefit, however, in the subgroup of patients who required urgent asthma care during the trial. Trial Registration clinicaltrials.gov Identifier: NCT00195117.


Functional analysis in vivo of engineered valved venous conduit with decellularized matrix and two bone marrow-derived progenitors in sheep.
Tissue engineering has been considered a promising approach for creating grafts to replace autologous venous valves. Here, ovine bone marrow-derived endothelial progenitor cells (EPCs) and multipotent adult progenitor cells (MAPCs) were harvested and then loaded into decellularized venous matrix to create tissue-engineered (TE) valved vein. Subsequently, the ovine femoral veins containing the valve were removed and replaced by TE grafts or acellular matrix only. The morphology and function were analysed for up to 1 year by ultrasonography, angiography, H&E staining and scanning electron microscopy (SEM). The differentiation of seeded cells was traced immunofluorochemically. The results showed that decellularized venous matrix could initially and feebly attract endogenous cells, but failed afterwards and were insufficient to restore valve function. On the contrary, the seeded cells differentiated into endothelial cells (ECs) in vivo and formed a monolayer endothelium, and smooth muscle cells within the scaffold therefore produced TE grafts comparable to the native vein valve. This TE graft remained patent and sufficient after implantation into the venous circuit of the ovine lower extremity for at least 6 months. Unfortunately, cells seeded on the luminal surface and both sides of the leaflets lost their biological functions at 12 months, resulting in thrombosis formation and leading to complete occlusion of the TE grafts and impotent venous valves. These findings suggest that this TE valved venous conduit can function physiologically in vivo in the medium term. Before translating this TE venous valve into clinical practice, the durability should be improved and thrombogenicity should be suppressed. Copyright © 2016 John Wiley & Sons, Ltd.


Association Between Oral Health and Frailty Among American Older Adults.
We examined the association between tooth loss, periodontal diseases, and frailty among older American adults. Data from the National Health and Nutrition Examination Surveys (NHANES) 2011-2014 was used. We included 2368 community-dwelling adults aged 60 years and older. Frailty was measured with the 49-item frailty index. Oral health indicators included number of teeth and periodontal disease. A composite nutritional intake variable based on 13 micronutrients from the dietary assessment was created. Negative binomial regression was used to test the association between oral health and frailty. The first model was adjusted for age and gender, the second model was additionally adjusted for nutritional intake, and the third model was additionally adjusted for other covariates. For each additional tooth, the rate ratio (RR) for frailty was 0.99 [95% confidence interval (CI) 0.98-0.99] in the fully adjusted model. Similarly, participants with moderate-severe periodontitis had 1.08 RR (95% CI 1.02-1.14) for frailty index compared with participants with no periodontitis after adjusting for age, gender, and poor nutritional intake. The association lost significance in the fully adjusted model. Oral health is associated with the frailty index, and nutritional intake appears to have a modest effect on the association. Periodontal disease has a weaker association with frailty compared with number of teeth. The findings highlight the importance of maintaining good oral health at older age and incorporating oral health indicators in routine geriatric assessments. Future research should investigate the role of potential mediating factors in this association.


Improving spot homogeneity by using polymer substrates in matrix-assisted laser desorption/ionization mass spectrometry of oligonucleotides.
We describe a method for improving the homogeneity of MALDI samples prepared for analysis of small, single-stranded oligonucleotides using the widely used DNA matrix system, 3-hydroxypicolinic acid/picolinic acid/ ammonium citrate. This matrix system typically produces large crystals around the rim of the dried sample and requires tedious searching of this rim with the laser. However, when a substrate is prepared using both Nafion and a hydrophilic, high-molecular-weight polymer, such as linear polyacrylamide, linear poly(ethylene oxide), or methyl cellulose, oligonucleotide-doped matrix crystals tend to be smaller and more uniformly distributed across the entire spot, thus decreasing the time that is required for locating a usable signal. In addition to MALDI characterization of the spatial distribution of "sweet spots," fluorescence microscopy allows for imaging dye-labeled DNA in dried MALDI spots. The mechanism of enhanced uniformity may involve increased viscosity in the MALDI sample droplet due to partial solubilization of the substrate by the MALDI sample solvent as well as partitioning of the matrix or DNA between the solvent and the undissolved portion of the polymer substrate.


Glacial refugia and the phylogeography of Steller's sea lion (Eumatopias jubatus) in the North Pacific.
Mitochondrial DNA sequence data were used to examine the phylogeographic history of Steller's sea lions (Eumetopias jubatus) in relation to the presence of Plio-Pleistocene insular refugia. Cytochrome b and control region sequences from 336 Steller's sea lions reveal phylogenetic lineages associated with continental refugia south of the ice sheets in North America and Eurasia. Phylogenetic analysis suggests the genetic structure of E. jubatus is the result of Pleistocene glacial geology, which caused the elimination and subsequent reappearance of suitable rookery habitat during glacial and interglacial periods. The cyclic nature of geological change produced a series of independent population expansions, contractions and isolations that had analogous results on Steller's sea lions and other marine and terrestrial species. Our data show evidence of four glacial refugia in which populations of Steller's sea lions diverged. These events occurred from approximately 60,000 to 180,000 years BP and thus preceded the last glacial maximum.


The Wnt/Ca2+ signaling pathway is essential for the regeneration of GABAergic neurons in planarian Dugesia japonica.
The growth and differentiation of neurons are critical events in the establishment of proper neuron connectivity and function. Planarians have a remarkable ability to completely regenerate a functional nervous system from a pluripotent stem cell population. Thus, planarians provide a powerful model to identify genes required for neuronal differentiation in vivo. The Wnt/Ca2+ signaling pathway is crucial for cancer development, arousing inflammatory responses, and neurodegeneration. We analyzed the expression patterns and RNAi phenotypes for members of the Wnt/Ca2+ signaling pathway in the planarian, Dugesia japonica. The expression of DjWnt5a, DjPLC-β, DjCamKII, and DjCaln during regeneration was surprisingly similar and revealing in the regenerated brain. RNAi knockdown of DjWnt5a, DjPLC-β, DjCamKII, and DjCaln led to defects in regenerated brains including brain partial deletions, incompact phenotypes at the posterior of the new brain, and lateral branches, which could not regenerate. Furthermore, the expressions of GAD and the number of GABAergic neurons decreased. Together, these results suggest that the Wnt/Ca2+ signaling pathway is required for GABAergic neuron regeneration.


Joint action of phenolic acid mixtures and its significance in allelopathy research.
Although the ecological significance of mixtures of phytotoxins is recognized in research on chemical plant interference (allelopathy), few studies convincingly demonstrate the joint action of phytotoxin mixtures, key to understanding the ecological impact of these materials, using established models from other biological disciplines, e.g. toxicology and pharmacology. Addressing this need, the present study investigates the joint action of the phenolic acids, p-hydroxybenzoic, p-coumaric and ferulic acids on root growth inhibition of perennial ryegrass (Lolium perenne L). The Additive Dose Model (ADM) isobole and estimated concentration of phenolic acid mixture were calculated on ED20, ED50 and ED80 from the dose-response curves for the phenolic acids applied alone or in mixtures of fixed ratios. The binary combination of three selected phenolic acids is generally antagonistic relative to the ADM. No evidence for synergistic activities of phenolic acids in the mixture was noted. Since allelopathic activities in nature are largely due to the presence of several compounds in a mixture, the present study advances understanding of the joint action of binary combination of allelochemicals in a mixture.


Risk profile and quality of dental restorations: a cross-sectional study.
The aims of the present study were (1) to evaluate the percentage of recurrent caries with respect to the estimated caries risk profile obtained with a Cariogram, (2) to evaluate the quality of restorations in a Saudi population with several restorations and (3) to determine the additional value of bite-wing radiographs as an aid to quality evaluation. A total of 803 restorations were examined in 100 adults according to the United States Public Health Service/Ryge criteria. Salivary and microbiological factors, dietary habits and plaque index were investigated. The Cariogram was used to evaluate the risk profiles. Class II bite-wing radiographs (n = 281) were taken to examine the marginal integrity and the anatomic form proximally. The patients were categorized according to 'the chance of avoiding caries' into three risk groups: 0%-20% (n = 38), 21%-40% (n = 28) and 41%-100% (n = 34). ANOVA revealed statistically significant differences between the risk groups with respect to the recurrent caries (P < 0.05). A high percentage of the total restorations (56%) were diagnosed with recurrent caries. The quality of anatomic form and surface texture was unacceptable in the majority of cases. After adding the evaluations of class II bite-wings, the percentage of unacceptable restorations increased by 28% and 17% with regard to marginal integrity and anatomic form, respectively (P < 0.001). Recurrent caries was related to the percentage 'chance of avoiding caries' as estimated by the Cariogram. The importance of bite-wings was emphasized as an aid to quality evaluation.


[Examination of endoscopical ultrasonography (EUS) on carcinoid tumors of gastrointestinal tract].
In order to clarify the usefulness of EUS in the diagnosis of gastrointestinal carcinoid tumor, we examined 15 patients, who had carcinoid tumors of gastrointestinal tract (stomach: 5, duodenum: 2, rectum: 8), on the diagnosis in quality and depth by comparing endoscopical ultrasonography (EUS) with resected specimen. Carcinoid tumors of gastrointestinal tract were detected as homogenous hypoechoic tumors with sharp border in all site by EUS. Especially, it was characteristic that 14 of the 17 lesions (82%) which invaded into submucosa were mainly located in the third layer, and the second layer covered the tumor at the foot, and near the top, it touched the tumor and got indistinct. We decided the depth of invasion by comparing the hypoechoic tumor with normal structure of 5 layers. The accuracy rate was 88%. In conclusion, EUS was thought to be useful in the diagnosis and choice for treatment of carcinoid tumors of gastrointestinal tract.


The cellulosome: an exocellular, multiprotein complex specialized in cellulose degradation.
Clostridium thermocellum produces a highly active cellulase system that consists of a high-M(r) multienzyme complex termed cellulosome. Hydrolytic components of the cellulosome are organized around a large, noncatalytic glycoprotein termed CipA that acts both as a scaffolding component and a cellulose-binding factor. Catalytic subunits of the cellulosome bear conserved, noncatalytic subdomains, termed dockerin domains, which bind to receptor domains of CipA, termed cohesin domains. CipA includes nine cohesin domains, a cellulose-binding domain, and a specialized dockerin domain. Proteins of the cell envelope carrying cohesin domains that specifically bind the dockerin domain of CipA have been identified. These proteins may mediate anchoring of the cellulosomes to the cell surface. Cellulase complexes similar to the cellulosome of C. thermocellum are produced by several cellulolytic clostridia. High-M(r) multienzyme complexes have also been identified in anaerobic rumen fungi. The architecture of the fungal complexes also seems to rely on the interaction of conserved, noncatalytic docking domains with a scaffolding component. However, the sequence of the fungal docking domains bears no resemblance to the clostridial dockerin domains, suggesting that the fungal and clostridial complexes arose independently.


Planar polarization of the denticle field in the Drosophila embryo: roles for Myosin II (zipper) and fringe.
Epithelial planar cell polarity (PCP) allows epithelial cells to coordinate their development to that of the tissue in which they reside. The mechanisms that impart PCP as well as effectors that execute the polarizing instructions are being sought in many tissues. We report that the epidermal epithelium of Drosophila embryos exhibits PCP. Cells of the prospective denticle field, but not the adjacent smooth field, align precisely. This requires Myosin II (zipper) function, and we find that Myosin II is enriched in a bipolar manner, across the parasegment, on both smooth and denticle field cells during denticle field alignment. This implies that actomyosin contractility, in combination with denticle-field-specific effectors, helps execute the cell rearrangements involved. In addition to this parasegment-wide polarity, prospective denticle field cells express an asymmetry, uniquely recognizing one cell edge over others as these cells uniquely position their actin-based protrusions (ABPs; which comprise each denticle) at their posterior edge. Cells of the prospective smooth field appear to be lacking proper effectors to elicit this unipolar response. Lastly, we identify fringe function as a necessary effector for high fidelity placement of ABPs and show that Myosin II (zipper) activity is necessary for ABP placement and shaping as well.


Optimization of mirror therapy to excite ipsilateral primary motor cortex.
Mirror illusion therapy (mirror therapy) is based on the experimental substrate of a visual illusion of active hand movement to excite ipsilateral primary motor cortex (iM1). We tested whether iM1 excitability could be modulated by enabling or disabling vision of the active hand during mirror therapy. Motor cortical activations of healthy right-handed participants were identified by magnetoencephalography. Participants performed voluntary index finger extension of their dominant and non-dominant hands, separately, while viewing mirror reflection images of their active hand superimposed upon their hidden inactive hand. This was performed either with vision of the active hand (uncovered viewing condition) or without vision of the active hand (covered viewing condition). In the covered viewing condition, the iM1could be excited in all participants (n = 10) and this excitation did not differ whether the active hand was the dominant or non-dominant hand. However, in the uncovered viewing condition, dominant and non-dominant hands were able to excite iM1 only in some participants (n = 4 and n = 7, respectively). Moreover, the participants' responses to the illusion validation questionnaire revealed that the covered viewing condition could cause clearer visual illusion for the active hand than the uncovered viewing condition. Disabling vision of the active hand during mirror therapy was more effective to excite iM1 responses by creating more immersive visual illusion of the active hand.


The G-protein coupled receptor family: actors with many faces.
G-protein coupled receptors (GPCRs) comprise the largest family of proteins in our body, which have many important physiological functions and are implicated in the pathophysiology of many serious diseases. GPCRs therefore are significant targets in pharmaceutical research. GPCRs share the common architecture of seven plasma membrane-spanning segments connected to each other with three extracellular and three intracellular loops. In addition, GPCRs contain an extracellular N-terminal region and an intracellular C-terminal tail. GPCRs could stimulate different intracellular G-proteins (internal stimuli) and signaling pathways after their interaction with different ligands (external stimuli). The exceptional functional plasticity of GPCRs could be attributed to their inherent dynamic nature to adopt different active conformations, which are stabilized differentially by different stimuli as well as by several mutations. This review describes the structural changes of GPCRs associated with their activation. Understanding the dynamic nature of GPCRs could potentially contribute in the development of future structure-based approaches to design new receptor-specific, signaling-selective ligands, which will enrich the pharmaceutical armamentarium against various diseases.


Probing Magnetism in the Vortex Phase of PuCoGa_{5} by X-Ray Magnetic Circular Dichroism.
We measure x-ray magnetic circular dichroism (XMCD) spectra at the Pu M_{4,5} absorption edges from a newly prepared high-quality single crystal of the heavy-fermion superconductor ^{242}PuCoGa_{5}, exhibiting a critical temperature T_{c}=18.7 K. The experiment probes the vortex phase below T_{c} and shows that an external magnetic field induces a Pu 5f magnetic moment at 2 K equal to the temperature-independent moment measured in the normal phase up to 300 K by a superconducting quantum interference device. This observation is in agreement with theoretical models claiming that the Pu atoms in PuCoGa_{5} have a nonmagnetic singlet ground state resulting from the hybridization of the conduction electrons with the intermediate-valence 5f electronic shell. Unexpectedly, XMCD spectra show that the orbital component of the 5f magnetic moment increases significantly between 30 and 2 K; the antiparallel spin component increases as well, leaving the total moment practically constant. We suggest that this indicates a low-temperature breakdown of the complete Kondo-like screening of the local 5f moment.


Using Ceramic-on-Ceramic Bearings in Total Hip Arthroplasty Necessitating 44- or 46-mm Metal Shells.
Ceramic-on-ceramic articulation shows the lowest wear, with a low incidence of osteolysis. A small cup limits options regarding liner thickness and femoral implant size. The purpose of this study was to evaluate the safety of a 4-mm-thick ceramic liner and the outcomes of cementless total hip arthroplasty using a 44- or 46-mm metal shell and a ceramic-on-ceramic bearing at mid- to long-term follow-up. Between May 2003 and June 2008, 80 patients (88 hips) who had hip osteoarthritis and a hypoplastic acetabulum underwent cementless total hip arthroplasty using a 44- or 46-mm metal shell and a ceramic-on-ceramic bearing. Sixty-eight of these patients (76 hips) were followed for an average of 8 years. The ceramic-related complications, clinical and radiological results, and Kaplan-Meier survivorship of these patients were evaluated. Mean Harris hip score had improved to 88 points (range, 49-100 points) at final follow-up. No ceramic fractures occurred during follow-up. All acetabular and femoral prostheses had bone ingrowth, and there was no measurable wear in any hip. In 1 patient, a radiolucent lesion occurred around the acetabular cup. Two hips underwent revision because of periprosthetic fractures. The survival rates of the acetabular cups and the femoral stems were 100% and 97.4%, respectively. The thin (4 mm) ceramic liner did not increase the risk of ceramic fracture and did not affect the mid- to long-term results and survival of cementless total hip arthroplasty using 44- or 46-mm metal shells. [Orthopedics. 2018; 41(3):e354-e359.].


[Berberine hydrochloride attenuates liver injury induced by acute hypoxic exposure by inhibiting TNF-α/caspase-8/caspase-3 signaling pathway].
Objective To investigate the protective effect and mechanism of berberine hydrochloride (BBR) on liver after acute hypoxic exposure. Methods C57BL/6 mice were divided into three groups consisting of normoxic group, hypoxic exposure group, and hypoxic exposure combined with BBR group. On the 7th day of the experiment, mice were sacrificed and liver tissue was collected. The pathological changes of liver tissue were observed by HE staining. The mRNA levels of interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α), and inducible nitric oxide synthase (iNOS) in liver tissue were detected by real time quantitative PCR. The protein levels of TNF-α, cleaved-caspase-3 (c-caspase-3), and caspase-8 in liver tissue were detected by Western blotting. The apoptosis of mouse liver tissue was detected by TUNEL. Results After 7 days of hypoxic exposure, the body mass and liver mass of mice decreased significantly, and the liver tissue damage was obvious; the mRNA levels of TNF-α and IL-1β and the protein levels of caspase-8 and c-caspase-3 in liver tissue cells significantly increased, and the apoptosis level of liver tissue cells markedly increased as well. BBR treatment significantly increased the body mass and liver mass of mice exposed to hypoxia for 7 days, decreased the mRNA level of TNF-α and the protein expressions of caspase-8 and c-caspase-3, and reduced the apoptosis of liver tissue cells. Conclusion BBR may attenuate liver injury induced by hypoxic exposure by inhibiting TNF-α/caspase-8/caspase-3 signaling pathway.


Quality of life in children after mild head injury.
The study was conducted to evaluate the effects of Mild Head Injury (MHI) in children not only in terms of impairment, but also in terms of disability, handicap and quality of life (QOL). Emergency Clinical Hospital "Bagdasar-Arseni", Bucharest, Romania, between 2000 and 2004 We take into account the patients with mild head injury MHI (CCS of 14 and 15 and amnesia). From a cohort of 1,319 children, consecutive patients with MHI, presented at the emergency room in a period of four years (2000-2003), 528 children (40.0%) were selected for admission, based on the presence of the risk factors. All admitted patients were investigated based on a protocol of neurosurgical evaluation and were followed for a period of 12 months. The Falls were the most common cause of MHI (30.6% - 162 cases). The proportion of children with detectable CT scan abnormalities was smaller (19.8% - 105 cases) and surgery was necessary in only 5.5% (29 cases). Special attention was paid to child-abuse and traffic accident cases. Post-concussion syndrome (PCS) was observed in 26.9% cases. Neuropsychological tests were performed in 96 children (21.2%), to evaluate neuropsychological, emotional, psychosocial and behavioral impairments. The study has shown that cognitive dysfunctions mainly were observed after MHI (especially deficits in information processing speed, memory and attention). The neurosurgeon should perform a complete evaluation of the children-patient with MHI, including a current physical examination, a neuro-radiological evaluation and a formal neuropsychological assessment, in order to detect the abnormalities and to treat them. Psychotherapy can be of benefit in cases with MHI. Any common case of MHI may hide a possible lesion with delayed consequences.


Direct liquid chromatography-tandem mass spectrometry determination of underivatized glyphosate in rice, maize and soybean.
The residue determination of the widely used herbicide glyphosate (GLY) is highly problematic due to its amphoteric character, low mass and lack of chemical groups that might facilitate its detection. Most methods developed up to now have employed pre-column or post-column derivatization to form fluorescent derivatives and/or to reduce the polar character of the analyte facilitating its chromatographic retention. The aim of this work is to evaluate the feasibility of performing the direct LC-MS/MS determination of GLY residues in vegetables. After testing several Hydrophilic Interaction Liquid Chromatography (HILIC) columns, Obelisc N was selected due to its better chromatographic retention. LC-MS/MS determination has been performed in negative ionization mode, monitoring up to four transitions to give high reliability to the identification/confirmation process. This approach has been evaluated for the determination of GLY residues in rice, maize and soybean samples, and the method validated at different concentrations in compliance with the maximum residue limits established in the current legislation. After sample extraction with water, a combination of extract dilution, partition with dichloromethane, and solid phase extraction (SPE) using Oasis HLB cartridges (depending on the sample matrix under analysis) was applied. Quantification was made by using isotope-labeled GLY as internal standard and calibration in solvent. The methodology developed allows the rapid determination of GLY residues avoiding the derivatization step typically applied for this herbicide. The most critical issue is the robustness of the Obelisc N column, which was found to suffer rapid degradation with time. Extreme care and continuous testing of retention times and peak shapes is required for a reliable determination.


[Nitrogen oxide in the treatment of oxygenation failure in neonates].
Oxygenation problems are common in very sick infants, and involve a high risk of mortality or residual morbidity. We describe two infants with severe oxygenation difficulties. In one baby these were caused by meconium aspiration, while in the second, premature (1,500 g birth weight) infant Escherichia coli septicemia had precipitated persistent pulmonary hypertension. Inhaled nitric oxide led to immediate improvement in oxygenation, and the one infant was weaned off mechanical ventilatory support after four days, the other after ten days. There were no discernible side effects in either baby. Nitric oxide treatment should be considered for infants with life-threatening oxygenation problems who do not respond adequately to conventional management.


Myringoplasty in children.
This series examines the results of myringoplasty in children using the overlay technique. Controversy still surrounds the issue of proper timing of surgery on the basis of preoperative assessment of tubal function. Results of this technique in children, using contralateral ear status as the primary predictor of success, found an overall success rate of 78%. Age appears to be a significant factor because children 10 years old and younger were found to have a higher incidence of surgical failures.


Effects of hypothalamic lesions on lymphocyte subsets in mice.
This study investigated the time course of the dynamics of lymphocyte subsets in the peripheral blood of mice, after bilateral lesions were made at anterior, middle and posterior sites in the hypothalamus of the mice, using a stereotaxic technique. Lymphocyte subsets were analyzed by using monoclonal antibodies specific to cell surface markers and a laser flow cytometry system. The results indicate that the anterior and posterior parts of the hypothalamus seem to play a role in immunomodulation of both helper and suppressor T lymphocytes. The middle part of the hypothalamus is considered to be the site of a control mechanism in the immune regulation of T and B lymphocytes.


Intraligand Charge Transfer Sensitization on Self-Assembled Europium Tetrahedral Cage Leads to Dual-Selective Luminescent Sensing toward Anion and Cation.
Luminescent supramolecular lanthanide edifices have many potential applications in biology, environments, and materials science. However, it is still a big challenge to improve the luminescent performance of multinuclear lanthanide assemblies in contrast to their mononuclear counterparts. Herein, we demonstrate that combination of intraligand charge transfer (ILCT) sensitization and coordination-driven self-assembly gives birth to bright EuIII tetrahedral cages with a record emission quantum yield of 23.1%. The ILCT sensitization mechanism has been unambiguously confirmed by both time-dependent density functional theory calculation and femtosecond transient absorption studies. Meanwhile, dual-responsive sensing toward both anions and cations has been demonstrated making use of the ILCT transition on the ligand. Without introduction of additional recognition units, high sensitivity and selectivity are revealed for the cage in both turn-off luminescent sensing toward I- and turn-on sensing toward Cu2+. This study offers important design principles for the future development of luminescent lanthanide molecular materials.


Dominant marine heterotrophic flagellates are adapted to natural planktonic bacterial abundances.
Grazing controls bacterial abundances and composition in many ecosystems. In marine systems, heterotrophic flagellates (HFs) are important predators. Assemblages of HFs are primarily formed by species still uncultured; therefore, many aspects of their trophic behaviour are poorly known. Here, we assessed the functional response of the whole assemblage and of four taxa grown in an unamended seawater incubation. We used fluorescently labelled bacteria to create a prey gradient of two orders of magnitude in abundance and estimated ingestion rates. Natural HFs had a half-saturation constant of 6.7 × 105 prey ml-1 , a value lower than that of cultured flagellates and within the range of marine planktonic bacterial abundances. Minorisa minuta was well adapted to low prey abundances and very efficient in ingesting bacteria. MAST-4 and MAST-7 were also well adapted to the typical marine abundances but less voracious. In contrast, Paraphysomonas imperforata, a typical cultured species, did not achieve ingestion rate saturation even at the highest prey concentration assayed. Our study, beside to set the basis for the fundamental differences between cultured and uncultured bacterial grazers, indicate that the examined predator taxa have different functional responses, suggesting that they occupy distinct ecological niches according to their grazing strategies and prey preferences.


In vitro Natural Killer Cell Immunotherapy for Medulloblastoma.
How the immune system attacks medulloblastoma (MB) tumors effectively is unclear, although natural killer (NK) cells play an important role in immune defense against tumor cells. Interactions between receptors on NK cells and ligands expressed by tumor cells are critical for tumor control by immunotherapy. In this study, we analyzed tumor samples from 54 MB patients for expression of major histocompatibility complex class I-related chains A (MICA) and UL16 binding protein (ULPB-2), which are ligands for the NK group 2 member D activatory receptor (NKG2D). The percentage of MICA and ULBP-2 positive cells was higher than 25% in 68% and 6% of MB patients, respectively. A moderate-high intensity of MICA cytoplasmic staining was observed in 46% MB patients and weak ULBP-2 staining was observed in 8% MB patients. No correlation between MICA/ULBP-2 expression and patient outcome was found. We observed that HTB-186, a MB cell line, was moderately resistant to NK cell cytotoxicity in vitro. Blocking MICA/ULBP-2 on HTB-186, and NKG2D receptor on NK cells increased resistance to NK cell lysis in vitro. However, HLA class I blocking on HTB-186 and overnight incubation with IL-15 stimulated NK cells efficiently killed tumor cells in vitro. We conclude that although NKG2D/MICA-ULBP-2 interactions have a role in NK cell cytotoxicity against MB, high expression of HLA class I can protect MB from NK cell cytotoxicity. Even so, our in vitro data indicate that if NK cells are appropriately stimulated, they may have the potential to target MB in vivo.


Targeting the 3CLpro and RdRp of SARS-CoV-2 with phytochemicals from medicinal plants of the Andean Region: molecular docking and molecular dynamics simulations.
Given the highly contagious nature of SARS-CoV-2, it has resulted in an unprecedented number of COVID-19 infected and dead people worldwide. Since there is currently no vaccine available in the market, the identification of potential drugs is urgently needed to control the pandemic. In this study, 92 phytochemicals from medicinal plants growing in the Andean region were screened against SARS-CoV-2 3 C-like protease (3CLpro) and RNA-dependent RNA polymerase (RdRp) in their active sites through molecular docking. The cutoff values were set from the lowest docking scores of the FDA-approved drugs that are being used to treat COVID-19 patients (remdesivir, lopinavir, and ritonavir). Compounds with docking scores that were lower than cutoff values were validated by molecular dynamics simulation with GROMACS, using root mean square deviation (RMSD), root mean square fluctuation (RMSF), radius of gyration (Rg), and intermolecular hydrogen bonds (H-bonds). Furthermore, binding free energies were estimated using the MM-PBSA method, and ADMET profiles of potential inhibitors were assessed. Computational analyses revealed that the interaction with hesperidin (theoretical binding energies, ΔGbind = -15.18 kcal/mol to 3CLpro and ΔGbind = -9.46 kcal/mol to RdRp) remained stable in both enzymes, unveiling its remarkable potential as a possible multitarget antiviral agent to treat COVID-19. Importantly, lupinifolin with an estimated binding affinity to 3CLpro higher than hesperidin (ΔGbind = -20.93 kcal/mol) is also a potential inhibitor of the 3CLpro. These two compounds displayed suitable pharmacological and structural properties to be drug candidates, demonstrating to be worthy of further research.Communicated by Ramaswamy H. Sarma.


Gadolinium-enhanced magnetic resonance imaging of meningeal carcinomatosis in colon cancer.
Meningeal carcinomatosis has been infrequently reported in colorectal carcinoma. Recently the value of gadolinium-enhanced magnetic resonance imaging (MRI) in the examination of this disease has been described. We report a 55-year-old man with meningeal carcinomatosis from a known colon cancer. In this case, a gadolinium-enhanced MRI of the brain showed enhancing dura and nodular lesions which were not detected by enhanced computed tomography. The gadolinium-enhanced MRI may serve as an indicator of leptomeningeal involvement and may be a useful test in addition to the evaluation of cerebrospinal fluid cytology and chemistry.


Suppression of retinal neovascularization by small-interference RNA targeting erythropoietin.
To observe the effect of inhibition of retinal neovascularization by small-interference RNA (siRNA) targeting erythropoietin (EPO). Three siRNAs against EPO were designed and synthesized. Then they were transfected to NIH/3T3 cells by liposomes. RT-PCR and Western blot were used to evaluate the efficacy of siRNA in attenuating EPO expression in NIH/3T3 cells. One-week-old C57BL/6J mice were exposed to 75 +/- 2% oxygen for 5 days, then they were returned to room air to induce retinal neovascularization. The siRNA type shown as most powerful in reducing EPO expression in vitro was intravitreally injected in the treatment group. Retinal neovascularization was evaluated by angiography with injection of fluorescein-dextran and quantification of neovascular proliferative retinopathy after 5 days in room air. Moreover, RT-PCR and immunoblot analysis were used to determine whether local administration of siRNA could affect the expression of EPO in murine retinas. Among the 3 designed siRNAs (named siEPO1-3), siEPO2 is the most efficient in inhibiting EPO expression. In this murine model of oxygen-induced retinopathy, retinal neovascularization in the eyes with siEPO2 injection was significantly reduced compared with that of the contralateral control eyes. Similarly, histological analysis indicates that the number of neovascular nuclei protruding into the vitreous cavity was decreased compared to the control eyes. Furthermore, the expression of EPO in the retinas injected with siEPO2 was dramatically decreased. siRNA against EPO could inhibit experimental retinal neovascularization by reducing EPO expression in the retinas of mice. It may provide a powerful and novel therapeutic tool for ischemia-induced retinal diseases.


Further treatment of decolorization liquid of azo dye coupled with increased power production using microbial fuel cell equipped with an aerobic biocathode.
A microbial fuel cell (MFC) incorporating a recently developed aerobic biocathode is designed and demonstrated. The aerobic biocathode MFC is able to further treat the liquid containing decolorization products of active brilliant red X-3B (ABRX3), a respective azo dye, and also provides increased power production. Batch test results showed that 24.8% of COD was removed from the decolorization liquid of ABRX3 (DL) by the biocathode within 12 h. Metabolism-dependent biodegradation of aniline-like compound might be mainly responsible for the decrease of overall COD. Glucose is not necessary in this process and contributes little to the COD removal of the DL. The similar COD removal rate observed under closed circuit condition (500 Ω) and opened circuit condition indicated that the current had an insignificant effect on the degradation of the DL. Addition of the DL to the biocathode resulted in an almost 150% increase in open cycle potential (OCP) of the cathode accompanied by a 73% increase in stable voltage output from 0.33 V to 0.57 V and a 300% increase in maximum power density from 50.74 mW/m(2) to 213.93 mW/m(2). Cyclic voltammetry indicated that the decolorization products of the ABRX3 contained in the DL play a role as redox mediator for facilitating electron transfer from the cathode to the oxygen. This study demonstrated for the first time that MFC equipped with an aerobic biocathode can be successfully applied to further treatment of effluent from an anaerobic system used to decolorize azo dye, providing both cost savings and high power output.


Rickettsial illnesses, a leading cause of acute febrile illness.
Rickettsial illnesses, comprising mainly spotted fever group, typhus group and scrub typhus, are vector-borne re-emerging or newly emerging febrile illnesses where humans are an accidental dead-end host. They are a major cause of non-malarial febrile illnesses among returned travellers. They commonly present as an acute febrile illness and carry a characteristic entry wound (eschar) or a discrete erythematous maculo-popular rash based on the organism and the region. The illness severity is mainly dependent on the virulence of the rickettsial organism and delay in the diagnosis is known to cause severe illness with multi-organ involvement carrying high mortality. Almost all rickettsial infections respond to anti-rickettsial antibiotics such as doxycycline within 48-72 hours. Awareness of rickettsial illnesses and their various clinical presentations helps in early diagnosis and institution of appropriate treatment and hence prevent morbidity and mortality.


The evolving pattern of war-related injuries from the Afghanistan conflict.
This is a retrospective study of 922 victims of the Afghanistan war admitted to Mekkah Mukarramah Surgical Hospital, a 120-bed hospital, from April 1987 to April 1989. The range and extent of war-related injuries were reviewed. The lower limbs were the most common site of injuries. The ratio of critical-area wounds (trunk, head, and neck) to extremity wounds was 0.17, which reflects the lack of early emergency medical care for the wounded. This ratio can be used to monitor and evaluate the efficiency of the evacuation plans in a war situation.