Variations in local adaptation of allopatric Fasciola hepatica to French Galba truncatula in relation to parasite origin.
Two French populations of Galba truncatula were subjected to experimental infections with Egyptian and French isolates of Fasciola sp. miracidia, originating from cattle and sheep, to compare characteristics of snail infections in allopatric and sympatric groups. All sampled Egyptian isolates were identified as Fasciola hepatica using microsatellite markers. Compared to snails infected with French miracidia, snail survival at day 30 post-exposure was significantly greater in the Egyptian groups, while prevalence of infection was significantly lower (in an Egyptian group infected with cattle-derived miracidia) or did not show any significant differences in the other three cases. The total number of metacercariae was significantly higher in the four Egyptian groups. However, snail population and the mammalian origin of F. hepatica had also a significant effect on this parameter. The dissection of snail cadavers showed a significantly higher number of free rediae in the Egyptian groups, even if snail population also had a significant effect on the redial burden. Both Egyptian isolates of F. hepatica could easily develop in French snails, causing a low mortality in snails and inducing a metacercarial production higher than that noted in sympatric infections. However, the mammalian origin of F. hepatica eggs and the quality of snail populations as intermediate hosts had to be taken into account for studying local adaptation in reason of their effects on this process.


A novel mechanism to generate metallic single crystals.
Generally, the evolution of metallic single crystals is based on crystal growth. The single crystal is either produced by growing a seed single crystal or by sophisticated grain selection processes followed by crystal growth. Here, we describe for the first time a fully new mechanism to generate single crystals based on thermo-mechanically induced texture formation during additive manufacturing. The single crystal develops due to two different mechanisms. The first step is a standard grain selection process due to directional solidification, leading to a pronounced fiber texture. The second and new mechanism bases on successive thermo-mechanically induced plastic deformations and texture formation in FCC crystals under compression. During this second step, the columnar grain structure transforms into a single crystal by rotation of individual grains. Thus, the single crystal forms step by step by merging the originally columnar grain structure. This novel, stress induced mechanism opens up completely new perspectives to fabricate single crystalline components and to accurately adjust the orientation according to the load.


Significance of screening antibodies to hepatitis B virus core antigen among Syrian blood donors.
Hepatitis B virus (HBV) transmission via hepatitis B surface antigen (HBsAg)-negative blood donors has been reported. While many countries have implemented screening antibodies to hepatitis B core antigen (anti-HBc) to further enhance transfusion safety, HBsAg is still the only obligatory HBV screening test of blood donors in Syria. To evaluate the significance of screening anti-HBc to reduce the risk of transfusion transmitted HBV infection in Syria. A cohort cross-sectional prospective study included 1939 healthy blood donors assigned at the blood transfusion center of Damascus University. All donors were tested for HBsAg and anti-HBc. HBsAg-negative, anti-HBc-positive sera were further tested quantitatively for antibodies to hepatitis B surface antigen (anti-HBs) and 'anti-HBc alone' sera were considered for HBV quantitative real time polymerase chain reaction (qPCR). Among 1913 HBsAg-negative donors, 215 (11·2%) were anti-HBc-positive including 125 anti-HBs high-positive and 59 anti-HBs low-positive donors. The remaining 31 donors were 'anti-HBc alone', five of which were HBV DNA-positive. Our results suggest including anti-HBc as an additional screening test for blood donors in Syria to reduce the risk of HBV transmission. As the most cost-effective measure, anti-HBc-positive donors should be tested quantitatively for anti-HBs and only donors with no or low (<100 IU L(-1) ) anti-HBs should be deferred.


Hospital management model for biological hazards: A qualitative content analysis study.
Biological hazards such as epidemic, pandemic, and bioterrorism are the most probable types of emergencies and disasters, which cause a high rate of mortality and morbidity. Hospitals have an important role in reducing the consequences of biological hazards. Given this, we set out the present study to design a model for managing biological hazards in hospitals. A qualitative content analysis study was performed using in-depth semistructured interviews with 40 experts. We selected participants through purposeful sampling; also, we interviewed experienced experts, policy makers, and managers in the field of hospital management regarding biological hazards. The study setting was Iran Ministry of Health and hospitals as well as other relevant organizations responsible for hospital management in biological hazards. We analyzed the data concurrently using the method recommended by Graneheim and Lundman. After collecting and analyzing the data obtained from the interviews, we extracted the dimensions of our model in the form of four themes, including mitigation, preparedness, response, and recovery. Based on the results of the present study, we believe that a comprehensive and practical model for hospital preparedness regarding management of biological hazards is essential. The model of hospital preparedness in biological hazards leads to an effective response and reduces the physical, psychological, social, and economic consequences of biological hazards.


Komrower Lecture. Congenital disorders of glycosylation (CDG): it's all in it!
Congenital disorders of glycosylation (CDGs) are due to defects in the synthesis of the glycan moiety of glycoproteins or other glycoconjugates. This review is devoted mainly to the clinical aspects of protein glycosylation defects. There are two main types of protein glycosylation: N-glycosylation and O-glycosylation. N-glycosylation generally consists of an assembly pathway (in cytosol and endoplasmic reticulum) and a processing pathway (in endoplasmic reticulum and Golgi). O-glycosylation lacks a processing pathway but is otherwise more complex. Sixteen disease-causing defects are known in protein glycosylation: 12 in N-glycosylation and four in O-glycosylation. The N-glycosylation defects comprise eight assembly defects (CDG-I) designated CDG-Ia to CDG-Ih, and four processing defects (CDG-II) designated CDG-IIa to CDG-IId. By far the most frequent is CDG-Ia (phosphomannomutase-2 deficiency). It affects the nervous system and many other organs. Its clinical expression varies from extremely severe to very mild (and thus probably underdiagnosed). The most interesting disease in this group is CDG-Ib (phosphomannose isomerase deficiency) because it is so far the only efficiently treatable CDG (mannose treatment). It has a hepatic-intestinal presentation. The O-glycosylation defects comprise two O-xylosylglycan defects (a progeroid variant of Ehlers-Danlos syndrome and the multiple exostoses syndrome) and two O-mannosylglycan defects (Walker-Warburg syndrome and muscle-eye-brain disease). All known CDGs have a recessive inheritance except for multiple exostoses syndrome, which is dominantly inherited. There is a rapidly growing group of putative CDGs with a large spectrum of clinical presentations (CDG-x). Serum transferrin iso-electrofocusing remains the cornerstone of the screening for N-glycosylation defects associated with sialic acid deficiency. Abnormal patterns can be grouped in to type 1 and type 2. However, a normal pattern does not exclude these defects. Screening for the other CDGs is much more difficult, particularly when the defect is organ- or system-restricted. The latter group promises to become an important new chapter in CDG. It is concluded that CDGs will eventually cover the whole clinical spectrum of paediatric and adult disease manifestations.


Workplace based Potentially Malignant Oral Lesions Screening among Tobacco Consuming Migrant Construction Site Workers in Chennai, South India: A Pilot Study.
Vulnerable population groups such as migrant workers are identified as emerging high-risk groups for oral cancer owing to the high prevalence of smokeless tobacco consumption. Premature deaths due to oral cancer can be prevented by screening the population with high tobacco consumption practices and detecting early reversible stages of oral mucosal cavity lesions and facilitating linkages for further care. To assess prevalence of potentially malignant oral mucosal cavity lesions among tobacco consuming migrant construction workers in sub-urban Chennai, India. A workplace based cross-sectional study design. A cross-sectional study was conducted at workplaces i.e., construction sites for screening potentially malignant oral mucosal cavity lesions among migrant workers across 23 construction sites of Chennai during September 2019 - February 2020. An onsite, group health education session was provided about the harms of tobacco use to the migrants. Data entered in MS Excel was analysed using SPSS and multivariate analysis was performed. Among 640 migrants included in the study, 411 (64.2%) were less than 30 years of age, 623 (97.4%) were from north-eastern states of India such as West Bengal, Bihar, Rajasthan, Uttar Pradesh, Jharkhand. A considerable size (272, 42.5%) could not read or write and 355 (55.4%) earn a monthly income of less than ten thousand rupees. Current tobacco users were 619 (96.7%), smokeless tobacco users (463, 72.34%), smokers (206, 32.2%) and dual users (52,8.12%). Inflammatory mucosal lesions in the oral cavity were 70.97% and more among smokeless tobacco users comparable to 22.58% among tobacco smokers and was significantly associated with up to 20 years of tobacco consumption. Prevalence of Potentially malignant oral lesions among smokeless tobacco using interstate migrant construction site workers is very high and need urgent interventions.


Possible involvement of phosphatidylinositol-specific phospholipase C related to pertussis toxin-sensitive GTP-binding proteins during adipocyte differentiation of 3T3-L1 fibroblasts: negative regulation of protein kinase C.
Insulin/dexamethasone/methylisobutylxanthine (hormones/IBMX) induce 3T3-L1 fibroblasts to differentiate into adipocytes. Our previous study suggested that pertussis toxin (IAP)-sensitive GTP-binding protein(s) (G-protein) is involved in the process of differentiation by hormones/IBMX, accompanied by c-fos induction. Northern blotting indicated that among the IAP-sensitive G-proteins, the levels of Gi2 alpha, Go alpha, and Gi3 alpha mRNA were decreased, increased and unchanged, respectively. Gi1 alpha was undetectable and IAP attenuated the decrease in Gi2 alpha mRNA level but did not affect the change in Go alpha mRNA level during the adipocyte differentiation. These results indicate that IAP-sensitive Gi2 alpha mRNA level is decreased during adipocyte differentiation. A combination of phosphatidylinositol-specific phospholipase C (PI-PLC) and IBMX induced c-fos expression in 3T3-L1 fibroblasts similar to that induced with hormones/IBMX. c-fos induced by both stimulators was also diminished by anti-inositolglycan antibody or anti-PI-PLC antiserum. Insulin stimulated the release of inositolproteoglycan and diacylglycerol from 3T3-L1 fibroblasts, which was suppressed by IAP treatment. These findings suggested that one of the pathways of adipocyte differentiation induced by hormones/IBMX occurs via the inositolglycan-specific PI-PLC cascade coupled to IAP-sensitive G-protein(s). Both activation of glycerophosphate dehydrogenase and stimulation of insulin-dependent 2-deoxyglucose uptake induced by hormones/IBMX were enhanced in protein kinase C-depleted cells exposed to phorbol 12-myristate 13-acetate (PMA), and attenuated in IAP-treated cells. The level of a 32P-labeled 52 kDa protein in plasma membrane fractions immunoprecipitated by anti-PI-PLC antiserum was increased by PMA stimulation, abolished in PMA-treated cells, and increased in IAP-treated cells. These findings suggest that protein kinase C phosphorylates PI-PLC, resulting in a decrease in PI-PLC activity related to the signal transduction pathway of adipocyte differentiation of 3T3-L1 fibroblasts.


Bariatric surgical outcomes.
This article describes the procedures that are performed for weight loss and characterizes the associated short-term success (operative safety, in-hospital morbidity/mortality) and long-term efficacy(weight loss, weight loss maintenance, postoperative complications). It discusses each category of procedure and reviews the current outcomes literature. It also addresses the technical challenges that are involved with the performance of each procedure and how these challenges may affect short and long-term outcomes. It concludes by comparatively analyzing the outcomes of the various bariatric surgical procedures and their respective roles in effectively managing the morbidly obese patient.


Negative implications of fragmented public perspectives toward local alcohol programs.
Strengthening local alcohol programs so as to emphasize primary prevention is an active concern among mental health professionals; yet interventions intended to achieve this aim have produced only modest shifts in program orientation in most communities. An analysis of data obtained from those active in local affairs in a small community, using Q-sort techniques, suggested that contrasting perspectives toward alcohol abuse as a social problem may stand as an unseen barrier to relevant change.


Reduced levels of circulating adhesion molecules in adolescents with early-onset psychosis.
It is suggested that neurodevelopmental abnormalities are involved in the disease mechanisms of psychotic disorders. Although cellular adhesion molecules (CAMs) participate in neurodevelopment, modulate blood-brain barrier permeability, and facilitate leukocyte migration, findings concerning their systemic levels in adults with psychosis are inconsistent. We examined plasma levels and mRNA expression in peripheral blood mononuclear cells (PBMCs) of selected CAMs in adolescents with early-onset psychosis (EOP) aged 12-18 years (n = 37) and age-matched healthy controls (HC) (n = 68). EOP patients exhibited significantly lower circulating levels of soluble platelet selectin (~-22%) and soluble vascular cell adhesion molecule-1 (~-14%) than HC. We found no significant associations with symptom severity. PSEL mRNA expression was increased in PBMCs of patients and significantly negatively correlated to duration of illness. These findings suggest a role for CAMs in the pathophysiology of psychotic disorders.


Are measurements of patient safety culture and adverse events valid and reliable? Results from a cross sectional study.
The association between measurements of the patient safety culture and the "true" patient safety has been insufficiently documented, and the validity of the tools used for the measurements has been questioned. This study explored associations between the patient safety culture and adverse events, and evaluated the validity of the tools. In 2008/2009, a survey on patient safety culture was performed with Hospital Survey on Patient Safety Culture (HSOPSC) in two medical departments in two geographically separated hospitals of Innlandet Hospital Trust. Later, a retrospective analysis of adverse events during the same period was performed with the Global Trigger Tool (GTT). The safety culture and adverse events were compared between the departments. 185 employees participated in the study, and 272 patient records were analysed. The HSOPSC scores were lower and adverse events less prevalent in department 1 than in department 2. In departments 1 and 2 the mean HSOPSC scores (SD) were at the unit level 3.62 (0.42) and 3.90 (0.37) (p < 0.001), and at the hospital level 3.35 (1.53) and 3.67 (0.53) (ns, p = 0.19) respectively. The proportion of records with adverse events were 10/135 (7%) and 28/137 (20%) (p = 0.003) respectively. There was an inverse association between the patient safety culture and adverse events. Until the criterion validity of the tools for measuring patient safety culture and tracking of adverse events have been further evaluated, measurement of patient safety culture could not be used as a proxy for the "true" safety.


Exacerbated post-infarct pathological myocardial remodelling in diabetes is associated with impaired autophagy and aggravated NLRP3 inflammasome activation.
Diabetes mellitus (DM) patients surviving myocardial infarction (MI) have substantially higher mortality due to the more frequent development of subsequent pathological myocardial remodelling and concomitant functional deterioration. This study investigates the molecular pathways underlying accelerated cardiac remodelling in a well-established mouse model of diabetes exposed to MI. Myocardial infarction in DM mice was established by ligating the left anterior descending coronary artery. Cardiac function was assessed by echocardiography. Myocardial hypertrophy and cardiac fibrosis were determined histologically 6 weeks post-MI or sham operation. Autophagy, the NLRP3 inflammasome, and caspase-1 were evaluated by western blotting or immunofluorescence. Echocardiographic imaging revealed significantly increased left ventricular dilation in parallel with increased mortality after MI in DM mice (53.33%) compared with control mice (26.67%, P < 0.05). Immunoblotting, electron microscopy, and immunofluorescence staining for LC3 and p62 indicated impaired autophagy in DM + MI mice compared with control mice (P < 0.05). Furthermore, defective autophagy was associated with increased NLRP3 inflammasome and caspase-1 hyperactivation in DM + MI mouse cardiomyocytes (P < 0.05). Consistent with NLRP3 inflammasome and caspase-I hyperactivation, cardiomyocyte death and IL-1β and IL-18 secretion were increased in DM + MI mice (P < 0.05). Importantly, the autophagy inducer and the NLRP3 inhibitor attenuated the cardiac remodelling of DM mice after MI. In summary, our results indicate that DM aggravates cardiac remodelling after MI through defective autophagy and associated exaggerated NLRP3 inflammasome activation, proinflammatory cytokine secretion, suggesting that restoring autophagy and inhibiting NLRP3 inflammasome activation may serve as novel targets for the prevention and treatment of post-infarct remodelling in DM.


Effect of contralateral and ipsilateral masking of acoustic stimulation on the latencies of auditory evoked potentials from cochlea and brain stem.
Masking techniques were applied to a group of 10 subjects with normal hearing, to investigate the influence of masking on the latencies of brain stem auditory evoked potentials. The levels of sufficient masking and overmasking were determined. Central masking, as established in a psychophysiological manner, cannot be clearly demonstrated by auditory brain stem evoked potentials. When masking the untested ear, the alterations in the response are small and this can be ignored in clinical practice if masking of the untested ear is necessary.


A clinical comparative study of thyroid surgeries with and without drain.
Most of the thyroid surgeries are accompanied by drain placement. The possibility of hematoma or seroma formation postoperatively is of concern to surgeons, as, thyroid is a highly vascular structure and a minor hemorrhage might also turn out to be life threatening, despite actual incidence of it being only 0.3-1%. Thus, drains are placed with an intention to obliterate dead space and clear the collecting blood or serum. Conversely, several studies have indicated chances of clots blocking the drainage tube and resulting in missing out of early detection of a massive bleed. Drains have also been known to add more discomfort to the patient, increase chances of infection, increase post operative pain, scar formation and longer duration of hospital stay. Thus, some surgeons believe in not placing a drain. To compare the outcomes of thyroid surgeries done with drain versus those thyroid surgeries done without drain. 40 patients, of either sex, between 18 and 70 years of age, who underwent thyroid surgeries for various thyroid disorders over a period of 6 months, were randomly allocated to either 'with drain' (group A) or 'without drain' (group B). The surgeon was made aware of drain status only intraoperatively. Patients were assessed for post-operative pain based on Visual Analogue Scale (VAS) at 6 and 24 h post-operatively. Post-operative complications including hematoma, seroma, and wound infection, if any, along with duration of hospitalization, were also documented. Patient was followed up one week post-operatively for wound check and suture removal. Both groups were homogenous according to age, sex, TIRADS, Bethesda, diagnosis and surgery performed. Postoperative pain at 6 and 24 h was significantly higher in group A than in group B [6.15 ± 1.31 vs 3.50 ± 0.88 (p = 0.001) (6 h), 4.45 ± 0.99 vs 1.20 ± 1.10 (p = 0.001) (24 h)]. Mean duration of hospitalization following thyroid surgery was significantly higher among group A than group B [3.80 ± 1.15 vs 2.15 ± 0.36 days (p = 0.001)]. Though not statistically significant, wound pain at 1 week and overall complications were higher in Group A than in Group B (p = 0.182, p = 0.127 respectively). Thyroid surgeries done without drain placement are likely to cause significantly lesser post-operative pain and shorter duration of hospitalization.


Study of the kinetics of the transport of Cu(II), Cd(II) and Ni(II) ions through a liquid membrane.
The coupled transport of Cu(II), Cd(II) and Ni(II) ions through a bulk liquid membrane (BLM) containing pyridine-2-acetaldehyde benzoylhydrazone (2-APBH) as carrier dissolved in toluene has been studied. Once the optimal conditions of extraction of each metal were established, a comparative study of the transport kinetics for these metals was performed by means of a kinetic model involving two consecutive irreversible first-order reactions. The kinetic parameters (apparent rate constants of the metal extraction and re-extraction reactions (k(1), k(2)), the maximum reduced concentration of the metal in the liquid membrane (R(o)max), the time of the maximum value of R(o)(t(max)) and the maximum entry and exit fluxes of the metal through the liquid membrane (J(f)max and J(s)max) of the extraction and stripping reactions were evaluated and results showed good agreement between experimental data and theoretical predictions. Complete transport through the membrane took place according to the following order: Cd(II)>Cu(II)>Ni(II), with similar kinetic parameters obtained for Cu(II) and Cd(III). The transport behaviour of Ni(II) was different to that of Cu(II) and Cd(III), probably due to the different stoichiometry of the nickel complex compared to those of the other metal ions and the different chemical conditions required for its formation. The influence of the sample salinity on the transport kinetics was studied. k(1) values decreased slightly when the feed solution salinity was increased for Cu(II) and Ni(II), but not for Cd(II). Values of k(2) were practically unaffected. The proposed BLM was applied to the preconcentration and separation of metal ions (prior to their determination) in water samples with different saline matrices (CRM, river water and seawater), and good agreement with the certified values was obtained.


Placebo responses in Parkinson's disease.
Parkinson's disease (PD) patients exhibit strong placebo responses in clinical trials. Patient characteristics that affect placebo include patients' expectations of good outcomes, genetic variants, and personality. The presence of motor fluctuation and high baseline UPDRS motor scores predicted placebo response. However, gender, age, duration of PD, religion, or level of education do not correlate with placebo response. PD patients who are preconditioned with active treatment such as apomorphine have more robust placebo effects. Studies that focused on patients with motor fluctuations, surgical intervention, or higher probability of placebo assignment had higher rates of placebo response. Patients view participating in placebo-controlled trials positively. Placebo effect can be measured objectively using neuroimaging and neurophysiological techniques. PET studies show that placebo-induced improvement is associated with dopamine release in the dorsal striatum and that the expectation of receiving the reward, not the reward itself, increased dopamine release in the ventral striatum. Expectations of benefitting from repetitive transcranial magnetic stimulation also induced dopamine release. Expectations of receiving a dopaminergic drug induced changes in fMRI in a reward-learning task. Single cell recordings demonstrate that placebo response is associated with changes of single neuronal activities in the basal ganglia circuit. These studies demonstrate that placebo effects are genuine biological responses to the administration of placebo. In clinical trials, we can use several approaches to minimize placebo responses. In clinical practice, we can use approaches to harness the power of placebo and minimize nocebo effects to improve patients' outcome.


Erythropoiesis and plasma tocopherol levels in irradiated mice.
Plasma protein and tocopherol concentrations, haematocrit and 59-iron incorporation into erythrocytes have been measured in vitamin E-deficient and supplemented mice before and after exposure to 500 R of 260 kVp X-ray. Supplemented animals had greater haematocrit, plasma tocopherol and protein levels initially. After irradiation plasma tocopherol concentration decreased drastically in the vitamin E-supplemented mice.


Coping flexibility and complicated grief: a comparison of American and Chinese samples.
The ability to process a death and the ability to remain optimistic and look beyond the loss are both thought to be effective means of coping with loss and other aversive events. Recently, these seemingly contrary dimensions have been integrated into the idea of coping flexibility. In this study, we assessed the ability of married and bereaved individuals in the United States and Hong Kong to use both coping approaches as operationalized by the trauma-focused and forward-focused coping scales of a previously validated questionnaire. We also calculated a single flexibility score. Bereaved participants reported greater trauma-focused coping ability than did married participants. However, bereaved participants meeting criteria for complicated grief (CG) reported less forward-focused coping than both asymptomatic bereaved and married participants. The CG group also showed less overall coping flexibility than the asymptomatic bereaved and married groups. Country was not a factor. Findings suggest that deficits in coping flexibility are indicative of pathology in bereaved individuals, and that this relationship extends across cultures. Limitations of the study and directions for future research are discussed.


Evaluation of diffusely high uptake by the calvaria in bone scintigraphy.
The present study was undertaken to evaluate the finding of diffusely high uptake by the calvaria in a series of 994 consecutive whole body scans in regard to the incidence and degree, and to discuss the mechanism. A total of 86 patients had positive scans (73 women and 13 men) and there were 71 malignant (27 breast, 16 lung, and 28 other malignancies), and 15 benign disease cases (5 degenerative bone, 3 blood, and 7 miscellaneous diseases). Positive rate of the finding was 8.7% in total, 14.9% in women, and 2.6% in men. The difference between women and men was significant (P less than 0.001). The finding was significantly correlative with the ages of patients. Also the finding is shown to be predominant in women ranging 50-59 years of age. On the other hand, the same finding as was made by Creutzig and his coworker was confirmed in malignant cases under high-dose anticancer drug therapy. On the basis of these results, the finding may be not only a side effect of intensive cytotoxic medication on the skeleton, but also a physiological event relative to bone change with age and postmenopausal osteoporosis.


Solvent effects on myoglobin conformational substates as studied by electron paramagnetic resonance.
Electronic paramagnetic resonance spectra of frozen horse myoglobin solutions at two different pH values and with different added organic solvents are analyzed by computer simulation in terms of Gaussian distributions of some ferric ion crystal field parameters. The mean values and the corresponding variances of these distributions, thought as arising from a distribution of the protein conformational substates, are found to be affected by both the pH and the addition of organic solvents. The significant narrowing of the conformational substate distribution, induced by large addition of glycerol, is discussed.


Protonated nucleobase ligands: synthesis, structure and characterization of 9-methyladeninium hexachloroplatinate and pentachloro(9-methyladeninium)platinum(IV).
Na(2)[PtCl(6)] was found to react with (9-MeAH)Cl(.)H(2)O (2) (9-MeA=9-methyladenine) in aqueous solution yielding (9-MeAH)(2)[PtCl(6)](.)2H(2)O (3). The same compound was obtained from hexachloroplatinic acid and 9-methyladenine. Performing this reaction at 60 degrees C, complex formation took place yielding the 9-methyladeninium complex [PtCl(5)(9-MeAH)](.)2H(2)O (4a). An analogous complex, [PtCl(5)(9-MeAH)](.)1/2(18C6)(.)H(2)O (4b, 18C6=crown ether 18-crown-6), was formed in the reaction of aquapentachloroplatinic acid (H(3)O)[PtCl(5)(H(2)O)](.)2(18C6)(.)6H(2)O (1) with 9-methyladenine in 1:1 ratio. All complexes were isolated in moderate to good yields as yellow powder (4b) and crystals (3, 4a), respectively. They were fully characterized by microanalysis, IR and NMR ((1)H, (13)C, (195)Pt) spectroscopies, and in part (2, 3, 4a) also by single-crystal X-ray diffraction analysis. Molecular structure of complex 4a exhibited that the 9-methyladeninium ligand is N1 protonated and coordinated through N7 to platinum(IV).


Trends in anemia treatment with erythropoietin usage and patient outcomes.
Recombinant erythropoietin, first approved for Medicare reimbursement in June 1989, was prescribed at initial doses for dialysis patients of 2,500 to 2,700 U per administration independent of hematocrit level. By 1997, however, patients with hematocrits less than 30% were administered 6,000 U/dose, compared with 4,500 U administered to patients with hematocrits of 33% to 36%. Since 1990, the percentage of patients with hematocrits less than 30% decreased from 60% to 22% in 1997, whereas the percentage of patients with hematocrits of 33% to 36% increased from 10% to 30%. In 1997, Medicare initiated the Hematocrit Measurement Audit (HMA) policy, which was directed at reducing the percentage of claims for hematocrits greater than 36% and increasing the stability of the hematocrit levels. The policy change achieved the initial effect but resulted in a reduction of the mean hematocrit as well. The policy was changed in 1998 in response to patient and provider concerns. Mortality studies show that hematocrits less than 30% (or hemoglobin levels < 110 g/L) are associated with an 18% to 40% increased associated risk for death. Higher hematocrits of 33% to 36% appear to be associated with a 7% reduced risk for death. The risk for hospitalization parallels that of mortality. Patients with sustained hematocrits of 33% to 36% over 1 year appear to have the best outcome compared with patients with hematocrits that decrease. The latter are at greater risk than those patients in whom the hematocrits increase. In conclusion, dramatic improvements in hemodialysis patient hematocrits have occurred since 1989. Mortality and hospitalization studies support the National Kidney Foundation Dialysis Outcomes Quality Initiative (NKF DOQI) target hematocrit range of 33% to 36% as providing the best associated outcomes.


D-sorbitol can keep the viscosity of dispersive ophthalmic viscosurgical device at room temperature for long term.
The combination of 3% sodium hyaluronate (HA) and 4% sodium chondroitin sulfate (CS) is used as a dispersive ophthalmic viscosurgical device (OVD) during cataract surgery. For most OVDs containing HA, storage at 2-8 °C is recommended to preserve product characteristics. In order to develop a dispersive OVD that can be stored at room temperature, in this study, we searched additives which can stably maintain the viscosity, a key parameter of OVD, under preservation stability testing at 60 °C. The addition of D-sorbitol to a combination OVD, 3% HA and 4% CS, suppressed the reduction in viscosity compared with other OVDs with or without additives. The addition of D-sorbitol was also effective in improving the residual viscosity of the combination OVD after thermal treatment and light irradiation. Moreover, the OVD containing D-sorbitol can be stored at 25 °C with stably maintaining the initial viscosity for at least 24 months. In conclusion, the new dispersive OVD, 3% HA, 4% CS, and 0.5% D-sorbitol, can be stored at room temperature instead of under cold conditions and may represent an attractive option for clinical use because it is not necessary to bring the product to room temperature prior to use.


A neomycin-resistant cell line for improved production of monoclonal antibodies to cell surface antigens.
Overgrowth of hybridomas by proliferating spleen T cells often hinders the production of monoclonal antibodies to certain antigens. To overcome this problem we derived neomycin-resistant fusion partner SP2 neoR.1 by transfection of SP2/0-Ag14 cells with plasmid pSVTK neo beta. Hybridomas obtained with SP2 neoR.1 grew optimally in the presence of the neomycin analog G418 at concentrations which blocked the proliferative response of T cells to mitogenic stimuli. The advantage of using SP2 neoR.1 and G418 under conditions where spleen cell proliferation occurs after fusion was demonstrated with hybridomas derived from a rat immunized with mouse helper T cell lines.


Intracardiac ultrasound-guided left bundle branch pacing in a bradycardia patient.
Left bundle branch pacing was associated with narrower QRS in patients with pacemaker indications. LBBP guided by ICE improves the accuracy of LBBP, facilitates the lead localization, minimizes complications, and above all reduces radiation exposure time.


Flow Sorting-Assisted Optical Mapping.
Optical mapping-a technique that visualizes short sequence motives along DNA molecules of hundred kilobases to megabase in size-has found an important place in genome research. It is widely used to facilitate genome sequence assemblies and analyses of genome structural variations. Application of the technique is conditional on availability of highly pure ultra-long high-molecular-weight DNA (uHMW DNA), which is challenging to achieve in plants due to the presence of the cell wall, chloroplasts, and secondary metabolites, just as a high content of polysaccharides and DNA nucleases in some species. These obstacles can be overcome by employment of flow cytometry, enabling a fast and highly efficient purification of cell nuclei or metaphase chromosomes, which are afterward embedded in agarose plugs and used to isolate the uHMW DNA in situ. Here, we provide a detailed protocol for the flow sorting-assisted uHMW DNA preparation that has been successfully used to construct whole-genome as well as chromosomal optical maps for 20 plant species from several plant families.


Dynamics of tumor oxygenation and red blood cell flux in response to inspiratory hyperoxia combined with different levels of inspiratory hypercapnia.
Increasing arterial oxygen partial pressure (pO2) by breathing hyperoxic gases is an effective means of improving tumor oxygenation, although the efficacy of adding CO2 to the inspiratory gas has been discussed controversially. This study aimed at analyzing the impact of different inspiratory CO2 fractions on the time course of oxygenation and perfusion changes in experimental tumors during and after inspiratory hyperoxia. Perfusion and oxygenation of rat DS-sarcomas were studied during spontaneous breathing of pure oxygen or hyperoxic gas mixtures containing different CO2 fractions (1, 2.5 or 5%). Red blood cell (RBC) flux was assessed as a measure of tumor perfusion using the laser Doppler technique and temporal changes in mean tumor pO2 were measured polarographically. Mean tumor pO2 increased 3.6-fold with pure oxygen, approx. 3.3-fold when 1 or 2.5% CO2 was added and 2.7-fold during carbogen breathing. RBC flux also increased by 25-30% with all gases. With pure oxygen and with 1% CO2 (+99% O2), perfusion changes paralleled those of the mean arterial blood pressure whereas with higher CO2 fractions, a decrease in resistance to flow was observed. The differences found with the various gas mixtures were more pronounced after the end of hyperoxia. With pure oxygen, perfusion immediately returned to pretreatment values whereas with higher CO2 fractions perfusion remained elevated for at least 30 min. Higher inspiratory CO2 fractions (2.5 or 5%) lead to a prolonged improvement of tumor perfusion after the end of inspiratory hyperoxia when compared with pure oxygen breathing. Since no principal differences in oxygenation and perfusion were seen between the gases containing 2.5 and 5% CO2, the former may be preferable for inspiratory hyperoxia.


A Novel Chiropteran Circovirus Genome Recovered from a Brazilian Insectivorous Bat Species.
This report describes the identification and characterization of a novel circovirus using metagenomic approaches in respiratory fluid samples from Brazilian free-tailed bats (Tadarida brasiliensis). The genome and deduced protein sequences share low identity with another circovirus recovered in distantly related bats from China.


A Material Conferring Hemocompatibility.
There is a need for biomimetic materials for use in blood-contacting devices. Blood contacting surfaces maintain their patency through physico-chemical properties of a functional endothelium. A poly(carbonate-urea) urethane (PCU) is used as a base material to examine the feasibility of L-Arginine methyl ester (L-AME) functionalized material for use in implants and coatings. The study hypothesizes that L-AME, incorporated into PCU, functions as a bioactive porogen, releasing upon contact with blood to interact with endothelial nitric oxide synthase (eNOS) present in blood. Endothelial progenitor cells (EPC) were successfully cultured on L-AME functionalized material, indicating that L-AME -increases cell viability. L-AME functionalized material potentially has broad applications in blood-contacting medical devices, as well as various other applications requiring endogenous up-regulation of nitric oxide, such as wound healing. This study presents an in-vitro investigation to demonstrate the novel anti-thrombogenic properties of L-AME, when in solution and when present within a polyurethane-based polymer.


Modulation of chaperone gene expression in mutagenized Saccharomyces cerevisiae strains developed for recombinant human albumin production results in increased production of multiple heterologous proteins.
The yeast Saccharomyces cerevisiae has been successfully established as a commercially viable system for the production of recombinant proteins. Manipulation of chaperone gene expression has been utilized extensively to increase recombinant protein production from S. cerevisiae, focusing predominantly on the products of the protein disulfide isomerase gene PDI1 and the hsp70 gene KAR2. Here we show that the expression of the genes SIL1, LHS1, JEM1, and SCJ1, all of which are involved in regulating the ATPase cycle of Kar2p, is increased in a proprietary yeast strain, developed by several rounds of random mutagenesis and screening for increased production of recombinant human albumin (rHA). To establish whether this expression contributes to the enhanced-production phenotype, these genes were overexpressed both individually and in combination. The resultant strains showed significantly increased shake-flask production levels of rHA, granulocyte-macrophage colony-stimulating factor, and recombinant human transferrin.


Spatiotemporal visualization of neuromagnetic data.
We describe the development of a computer visualization tool that displays topographic maps of neuromagnetic data on realistic head shapes. Two topographic display modes are implemented--a raw mode that depicts the actual position, orientation, and corresponding magnetic field data of each detection coil with respect to the head shape, and an interpolated mode, where the magnetic data are smoothly interpolated over the scalp. False coloration is used to represent magnetic field strength. The topographic displays are coupled with wave form displays in order to integrate spatial and temporal visualization of the data. The X Window system is used to write device independent, network-transparent computer code.


Layered hydrogels accelerate iPSC-derived neuronal maturation and reveal migration defects caused by MeCP2 dysfunction.
Probing a wide range of cellular phenotypes in neurodevelopmental disorders using patient-derived neural progenitor cells (NPCs) can be facilitated by 3D assays, as 2D systems cannot entirely recapitulate the arrangement of cells in the brain. Here, we developed a previously unidentified 3D migration and differentiation assay in layered hydrogels to examine how these processes are affected in neurodevelopmental disorders, such as Rett syndrome. Our soft 3D system mimics the brain environment and accelerates maturation of neurons from human induced pluripotent stem cell (iPSC)-derived NPCs, yielding electrophysiologically active neurons within just 3 wk. Using this platform, we revealed a genotype-specific effect of methyl-CpG-binding protein-2 (MeCP2) dysfunction on iPSC-derived neuronal migration and maturation (reduced neurite outgrowth and fewer synapses) in 3D layered hydrogels. Thus, this 3D system expands the range of neural phenotypes that can be studied in vitro to include those influenced by physical and mechanical stimuli or requiring specific arrangements of multiple cell types.


Expression of ubiquitin-conjugating enzyme E2C/UbcH10 in astrocytic tumors.
UbcH10 is one of the key regulators of cell cycle progression through the mitotic spindle assembly checkpoint pathway. Recently, aberrantly high UbcH10 expression has been demonstrated in a variety of malignancies. However, its role in astrocytic carcinogenesis is not well defined. This study investigated the splice pattern of the UbcH10 gene and its expression status in astrocytomas of different grades. Consequently, UbcH10 splice variant 1 (GenBank accession nos. NM_007019) was detected in astrocytomas and normal brain tissues by RT-PCR and sequence analysis. Expression levels of UbcH10 mRNA were elevated in high- versus low-grade astrocytomas (64.33+/-60.98 vs 8.36+/-8.15, respectively; p=0.000) or normal controls (64.33+/-60.98 vs 1.00+/-1.57, respectively; p=0.000), as determined by quantitative real time PCR analysis. Similarly, immunohistochemistry study showed increased UbcH10 labelling index in high-grade astrocytomas versus low-grade tumors (10.53+/-5.79% vs 4.23+/-2.85%, respectively; p=0.000) or normal controls (10.53+/-5.79% vs 0.0+/-0.0%, respectively; p=0.000) and, a positive correlation between UbcH10 immunoreactivity and Ki-67 immunostaining was also noted (Spearman r=0.63, p<0.001). These data suggest that overexpression of UbcH10 may serve as one important molecular mechanism that underlies the astrocytic carcinogenesis.


MicroRNA-186 induces sensitivity of ovarian cancer cells to paclitaxel and cisplatin by targeting ABCB1.
Recent studies have shown that microRNAs may regulate the ABCB1 gene (ATP-binding cassette, sub-family B [MDR/TAP], member 1). Computational programs have predicted that the 3'-untranslated region (3'-UTR) of ABCB1 contains a potential miRNA-binding site for miR-186. Here, we investigated the role of miR-186 in sensitizing ovarian cancer cells to paclitaxel and cisplatin. Human ovarian carcinoma cell lines OVCAR3, A2780, A2780/DDP, and A2780/Taxol were exposed to paclitaxel or cisplatin with or without miR-186 transfection, and cell viability was determined by MTT assay. Reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis were used to assess the MDR1, GST-π, and MRP1 expression levels. Dual-luciferase reporter assay was used to reveal the correlation between miR-186 and ABCB1. Lower miR-186 while higher MDR1 and GST-π mRNA expression levels were found in the A2780/Taxol and A2780/DDP cells than in the A2780 cells. After miR-186 transfection, all the cell lines showed increased sensitivity to paclitaxel and cisplatin. MiR-186 transfection induced apoptosis while anti-miR-186 transfection reduced apoptosis. The dual-luciferase reporter assay verified that that miR-186 combined with the 3'-untranslated region (UTR) of ABCB1. MDR1 and GST-π mRNA and protein expression levels were downregulated after transfection with miR-186 but upregulated following anti-miR-186 transfection compared to the mock and negative control cancer cells; however, the MRP1 expression levels did not significantly differ among the groups. Our results are the first to demonstrate that miR-186 may sensitize ovarian cancer cell to paclitaxel and cisplatin by targeting ABCB1 and modulating the expression of GST-π.


Modified technique for the exposure of the inferior alveolar nerve in rats.
The inferior alveolar nerve (IAN), a branch of the mandibular division of the trigeminal nerve, is a major component of the neurovascular bundle along with the inferior alveolar artery and vein. In rats, when exposed using an external oral approach while remaining intact, it can serve as an important tool to study the different effects of neuromediators and assess the role of different groups of nerve fibers. This paper describes a new technique to expose this nerve giving some experimental results to support its usefulness.


The Motor-Proficiency-Test for children between 4 and 6 years of age (MOT 4-6): an investigation of its suitability in Greece.
Given the negative influence of motor difficulties on people's quality of life their early identification seems to be crucial and consequently the information provided by a sound assessment tool is of great importance. The aim of this study was to examine the suitability of the MOT 4-6 (Zimmer & Volkamer, 1987) for use with preschoolers in Greece. Seven hundred and seventy-eight Greek children aged 48-71 months participated in the study. The two-way ANOVA used on total MOT performance revealed significant differences among the age groups formed in preschool age within Greeks, while boys' and girls' scores were quite similar. From the comparisons of Greeks' scores with the German standardization sample's ones, statistically significant differences were found in two age groups. However according to the Cohen's d effect size they were not of great importance. The distribution of Greeks' scores according to the test cut-offs, revealed that the MOT can differentiate all levels of performance, although a slight deviation from the distribution of Germans' scores was noticed. Finally, both the test-retest reliability and internal consistency of the test were found to be excellent. The MOT 4-6 seems to be a valuable motor assessment tool for Greek preschoolers. Regarding its norms, despite the minor differences that were noticed between the motor development of Greek and German preschoolers, their adjustment was thought to be unnecessary. Instead of lowering the norms, efforts for preventing the motor performance decline should be enhanced.


Toward sustainable culture media: Using artificial intelligence to optimize reduced-serum formulations for cultivated meat.
When considering options for future foods, cell culture approaches are at the fore, however, culture media to support the process has been identified as a significant contributor to the overall global warming potential (GWP) and cost of cultivated meat production. To address this issue, an artificial intelligence-based approach was applied to simultaneously optimize the GWP, cost, and cell growth rate of a reduced-serum culture media formulation for a zebrafish (ZEM2S cell line) cultivated meat production system. Response surface methodology (RSM) was used to design the experiments, with seven components - IGF, FGF, TGF, PDGF, selenium, ascorbic acid, and serum - selected as independent variables, given their influence on culture media performance. Radial basis function (RBF) neural networks and genetic algorithm (GA) were applied for prediction of dependent variables, and optimization of the culture media formulation, respectively. The results indicated that the developed RBF could accurately predict the GWP, cost and growth rate, with a model efficiency of 0.98. Subsequently, the three developed RBF neural networks predictive models were used as the inputs for a multi-objective genetic algorithm, and the optimal quantities of the independent variables were determined using a multi-objective optimization algorithm. The suggested RSM + RBF + GA framework in this study could be applied to sustainably optimize serum-free media development, identifying the combination of media ingredients that balances yield, environmental impact, and cost for various cultivated meat cell lines.


Compartmentalized immune responses: antigen-specificity of cerebrospinal fluid T-cell lines maintained in the absence of antigen.
The generation of long-term interleukin 2-dependent T-cell lines from anatomically compartmentalized sites of pathology offers a unique approach to the investigation of certain autoimmune diseases. However, it is generally believed that antigen-specific T-cell lines and clones lose antigen reactivity and specificity when propagated in the absence of antigen. Therefore, the optimal application of this approach to such diseases in which the pathogenetic antigens are unknown may be difficult. In approaching this problem, we have recently demonstrated that a proportion of antigen-specific T-cell lines derived from the peripheral circulation can maintain antigen specificity if propagated with antigen-presenting cells alone or with antigen-presenting cells together with OKT3 antibody, but in either case in the absence of antigen. In this report we describe the use of this approach to maintain the antigen specificity of T cells obtained from an anatomically compartmentalized site of pathology--the cerebrospinal fluid from a patient with tuberculous meningitis. We report here that a proportion of the T-cell lines generated from such cerebrospinal fluid lymphocytes can be maintained as antigen specific in the absence of antigen if propagated with either antigen-presenting cells alone or with antigen-presenting cells and OKT3 antibody. The approach illustrated in this report should now find broad applicability in the investigation of a number of autoimmune disease.


Epidermal growth factor (EGF)-induced up-regulation and agonist- and antagonist-induced desensitization and internalization of A1 adenosine receptors in a pituitary-derived cell line.
This report concerns the study of homologous and heterologous regulation of cell surface A1 adenosine receptors (A1R) in a pituitary-derived cell line. This has been possible by the use of the recently developed anti-A1R antibodies in immunocytochemical assays. Functional desensitization and internalization of A1R in GH4 cells occurred after treatment with agonist but also with antagonist. Epidermal growth factor (EGF) treatment led to the up-regulation of cell surface A1R in GH4 cells. Confocal analysis evidenced an EGF-induced increase of A1R present in intracellular clathrin-coated vesicles. The up-regulation was blocked by actinomycin D thus suggesting the involvement of protein synthesis in the effect induced by the growth factor. These results constitute the first example of adenosine receptor regulation by EGF and one of the few examples of antagonist-induced desensitization and internalization among G-protein-coupled receptors.


Predictors of the accuracy of self assessment of everyday functioning in people with schizophrenia.
Despite multiple lines of evidence suggesting that people with schizophrenia have substantial problems in self-reporting everyday functioning and cognitive performance, self-report methods are still widely used to assess functioning. This study attempted to identify predictors of accuracy in self report, both in terms of accurate self-assessment and over-estimation of current functioning. As part of the larger Validating Assessments of Everyday Real-World Outcomes (VALERO) study, 195 patients with schizophrenia were asked to self report their everyday functioning with the Specific Levels of Functioning (SLOF) scale, which includes subscales assessing social functioning, everyday activities, and vocational functioning. They were also assessed with measures of neuropsychological (NP) performance and functional capacity (FC), and were assessed for psychiatric symptomatology. In addition, a friend, relative or clinician informant was interviewed with the SLOF, and an interviewer with access to all information provided by the patient and informant (exclusive of performance-based data) generated "best estimate" ratings of actual, everyday functioning. Patients significantly (p<.001) overestimated their vocational functioning and everyday activities compared to the interviewer judgments. Lower levels of NP and FC performance and everyday functioning on the part of patients were consistently associated with overestimation of their functioning. Patient self-reports were not correlated with any performance-based measures, while interviewer judgments were significantly correlated with patients' performance on NP and FC measures (p<.005). In regression analyses, adjusting for interviewer ratings of functioning, several predictors of the discrepancy between self and interviewer judgments emerged. Higher levels of depressive symptoms were associated with less overestimation in self-reports (p<.001). Delusions, suspiciousness, grandiosity and poor rapport were all significantly (p<.001) associated with over-estimation of functioning compared to interviewer judgments. Poorer NP and FC performance were also associated with over-estimation of everyday functioning, but these results were not statistically significant in multivariate regression models. Consistent with previous studies in schizophrenia, other neuropsychiatric conditions and non-clinical populations, higher levels of depression were associated with increased accuracy in self-assessment. Similarly, lower scores on performance-based measures and judgments of everyday functioning also predicted over-estimation of functioning. Thus, we identified bi-directional predictors of mis-estimation of everyday functioning, even when poor baseline scores were considered. These data suggest that it may be possible to screen patients for their ability to self-report their functioning, but that performance-based measures of functioning provide a less biased assessment.


Improved technique for electrophoresis of human galactose-1-p uridyl transferase (EC 184.108.40.206).
A newly developed electrophoretic technique for human galactose-1-phosphate uridyl transferase confirms the multiple band patterns for the Duarte and Los Angeles variants. This represents the first confirmation for the Los Angeles variant. The observed frequencies of N, D, and LA types are similar to earlier reports for these variants.


Natural isotope correction improves analysis of protein modification dynamics.
Stable isotope labelling in combination with high-resolution mass spectrometry approaches are increasingly used to analyze both metabolite and protein modification dynamics. To enable correct estimation of the resulting dynamics, it is critical to correct the measured values for naturally occurring stable isotopes, a process commonly called isotopologue correction or deconvolution. While the importance of isotopologue correction is well recognized in metabolomics, it has received far less attention in proteomics approaches. Although several tools exist that enable isotopologue correction of mass spectrometry data, the majority is tailored for the analysis of low molecular weight metabolites. We here present PICor which has been developed for isotopologue correction of complex isotope labelling experiments in proteomics or metabolomics and demonstrate the importance of appropriate correction for accurate determination of protein modifications dynamics, using histone acetylation as an example.


Ritlecitinib: First Approval.
Ritlecitinib (LITFULO™), an orally administered kinase inhibitor, is being developed by Pfizer for the treatment of alopecia areata, vitiligo, ulcerative colitis and Crohn's disease. On 23 June 2023, ritlecitinib received approval in the USA for the treatment of severe alopecia areata in adults and adolescents 12 years and older. Ritlecitinib was approved in Japan on 26 June 2023 for the treatment of alopecia areata (limited to intractable cases involving widespread hair loss). Ritlecitinib has also received a positive opinion in the EU and is under regulatory review in the UK and China. This article summarizes the milestones in the development of ritlecitinib leading to this first approval for severe alopecia areata.


Perchlorate, thiocyanate, and nitrate in edible cole crops (Brassica sp.) produced in the lower Colorado River region.
The Colorado River is contaminated with low levels of perchlorate. Perchlorate has the potential to disrupt thyroid function by inhibiting the uptake of iodide. Brassica are rich sources of thiocyanate and nitrate, also inhibitors of iodide uptake. This study was conducted to estimate potential human exposure to perchlorate, thiocyanate, and nitrate from Brassica sp. irrigated with Colorado River water. Results indicate that Brassica sp. irrigated with Colorado River water do accumulate trace levels of perchlorate. However, the levels of perchlorate observed are low relative to the nitrate and thiocyanate naturally present in these species and low relative to the reference dose recommended by the NAS and the USEPA.


Pre-clinical evaluation of APrevent® VOIS for unilateral vocal fold paralysis medialization.
To evaluate the concept and efficacy of an adjustable implant (Prototype SH30: porcine implant and APrevent® VOIS: human concept) for treatment of unilateral vocal fold paralysis (UVFP) via in vivo mini-pig studies, human computed tomographic (CT) and magnetic resonance (MR) image analysis, ex-vivo aerodynamic and acoustic analysis. Feasibility testing and prototype implantation were performed using in-vivo UVFP porcine model (n = 8), followed by a dimensional finding study using CT and MR scans of larynges (n = 75) for modification of the implant prototypes. Acoustic and aerodynamic measurements were recorded on excised canine (n = 7) larynges with simulated UVFP before and after medialization with VOIS-Implant. The prototype showed in the in-vivo UVFP porcine model an improved glottic closure from grade 6 incomplete closure to complete closure (n = 5), to grade 2 incomplete closure (n = 2) and grade 3 incomplete closure (n = 1). On human CT/MR scans the identification of the correct size was successful in 97.3% using the thyroid cartilage alar "distance S" as the only parameter, which is an important step towards procedure standardization and implant design. Results were confirmed with implantation in human laryngeal cadavers (n = 44). Measurements of the acoustic and aerodynamic effects after implantation showed a significant decreased phonation threshold pressure (p = .0187), phonation threshold flow (p = .0001) and phonation threshold power (p = .0046) on excised canine larynges with simulated UVFP. Percent jitter and percent shimmer decreased (p = .2976; p = .1771) but not significant. Based on the preclinical results four sizes, differing in medial length, implant width and expansion direction of silicone cushions, seem to be enough to satisfy laryngeal size variations. This concept is significantly effective in medializing UVFP and improving the aerodynamic and acoustic qualities of phonation as reported in a preliminary clinical outcome study with long-term implantation. N/A.


Exercise and epilepsy: a survey of Midwest epilepsy patients.
Obesity and lack of physical activity are an increasing problem. In addition to common barriers to physical activity, people with epilepsy also face fear of exercise-induced seizures, medication side effects, and, often, confusing advice regarding the safety of exercise. To explore barriers faced by people with epilepsy, we mailed a survey to 412 adult patients with epilepsy from an epilepsy center in Kansas. Survey items assessed patients' exercise habits, attitudes regarding exercise, and barriers to exercise. Forty-seven percent completed the survey. Most respondents reported that they did exercise, though most did so 3 or fewer days per week and at light intensity. Respondents who reported seizures as a barrier to exercise did not exercise with less frequency or intensity than those who did not report seizures as a barrier, but these respondents reported greater fear of seizures. Patient-specific education about the benefits of exercise needs to be initiated by physicians.


Inhibitory effects of resveratrol derivatives from dipterocarpaceae plants on tyrosinase activity.
Stilbene derivatives, which are resveratrol (3,4',5-trihydroxy-trans-stilbene) oligomers ranging from monomer to tetramer, isolated from Dipterocarpaceae plants were tested for their inhibitory effects against murine tyrosinase activity. The structure-activity relationships obtained in this study suggest that the double bond in the stilbene skeleton is critical for the inhibition, and also that molecular size is important for inhibitory potency.


ATP formation onset lag and post-illumination phosphorylation initiated with single-turnover flashes. I. An assay using luciferin-luciferase luminescence.
The great sensitivity of the luciferin-luciferase ATP detection system allows direct observation of ATP formation derived from single-turnover flashes in a thylakoid reaction mixture. The method can measure the energization threshold--the number of flashes required for the initiation of ATP formation--as well as detect post-illumination ATP formation after the last flash of a flash sequence. We describe the characteristics of this post-illumination phosphorylation which can be observed after a series of phosphorylating flashes (PIP+) or when the assay for ATP formation was performed in a "traditional" manner where the ADP and Pi were added after the flash-energization period (PIP-). Comparing PIP+ yields and kinetics of the PIP+ decay under various treatments can give information about membrane energization events only if it is clearly established that different PIP+ yields and decay rates are not due to limitations of the luciferase-catalyzed reaction. Experiments showing that the PIP+ ATP yield and kinetics were due to membrane-limited deenergization events (proton efflux) rather than luciferase limitations include: (1) An uncoupler, nigericin, added after the last flash reduced the PIP+ yield, but had no effect on the luciferase reaction. (2) The kinetics of the luminescence after adding standard ATP were much faster than the PIP+ kinetics. (3) Valinomycin and K+ stimulated the PIP+ yield but had no influence on the luciferase reaction. (4) Lowering the pH from 8 to 7 increased both the PIP- (an assay independent of luciferase kinetics) and the PIP+ ATP yields, an expected result owing to the greater endogenous buffering power encountered by the proton gradient when the external pH is 7. In spite of the last-mentioned point, the threshold flash number for ATP formation onset was the same for pH 7 and 8 (valinomycin, K+ present) at slow flash frequencies. This is consistent with a membrane-localized rather than a delocalized gradient. The accompanying reports (W. A. Beard, G. Chiang and R. A. Dilley, and W. A. Beard and R. A. Dilley, J. Bioenerg. Biomembr.) show that different conditions can lead to observing either localized or delocalized proton gradient coupling in the PIP+ event and the ATP onset threshold flash number.


Amplification and overexpression of epidermal growth factor receptor gene in glioblastomas of Chinese patients correlates with patient's age but not with tumor's clinicopathological pathway.
It is believed that there are two distinct pathological pathways leading to the development of human glioblastomas (GBM) in Caucasian populations. Primary (de novo) GBM most often occurs in older individuals, and is characterized by the overexpression/amplification of epidermal growth factor receptor gene (EGFR), whereas secondary GBM, which progresses from a low-grade astrocytoma, often affects younger individuals and frequently contains the TP53 mutation. We and others have previously found that the age of onset of GBM in Chinese patients tends to be younger than that in Caucasian patients. To identify whether GBMs from Chinese patients share this common pattern of genetic alterations, expression levels of EGFR and TP53 and TP53 mutation were analyzed in 56 randomly selected Chinese GBMs (30 primary and 26 secondary), including 47 adult-onset and 9 pediatric GBMs. Consistent with other studies, overexpression/mutation of TP53 and aneuploid DNA content were more frequently detected in secondary GBMs of Chinese adult patients. In contrast to that observed in Caucasian patients, no significant difference was observed in the age distribution and the frequency of EGFR overexpression/amplification between primary and secondary GBMs in adult Chinese patients. Furthermore, the overexpression of EGFR was much higher in late-onset (age >45 years) GBMs (73%) than that in both early-onset (age 18-45 years) (17%) and pediatric (age <18 years) GBMs (11%), suggesting that overexpression of EGFR in Chinese GBMs may be associated closely with the patients age but not with the tumors' pathological pathway.


A liquid-liquid extraction procedure followed by a low temperature purification step for the analysis of macrocyclic lactones in milk by liquid chromatography-tandem mass spectrometry and fluorescence detection.
In this work a method is proposed and demonstrated for the analysis of the macrocyclic lactones abamectin, doramectin, eprinomectin, ivermectin and moxidectin in bovine milk by liquid chromatography coupled to mass spectrometry (LC-MS/MS) and liquid chromatography with fluorescence detection (LC-FL). The method is based on liquid-liquid extraction followed by a low temperature purification (LLE-LTP) step. Moreover, the proposed method was validated according to the Commission Decision 2002/657/EC, using LC-MS/MS and LC-FL for confirmatory and quantitative analysis, respectively. For LC-MS/MS the recovery rates observed ranged from 101.2 to 141.6% with coefficient of variation from 2.6 to 19.8%. For LC-FL the recovery rates observed ranged from 100.2 to 105% and coefficient of variations from 2.9 to 8.8%. Matrix effects were negligible due to the low temperature purification step. The quantification limits were far below the maximum limits established by regulations of all countries consulted. The proposed method proved to be simple, easy, and adequate for high-throughput analysis of a large number of samples per day at low cost.


DNA Barcoding for Identification of Sugarcane Borers in China.
Sugarcane borers are economically damaging insects with species that vary in distribution patterns both geographically and temporally, and vary based on ecological niche. Currently, identification of sugarcane borers is mostly based on morphological characters. However, morphological identification requires taxonomic expertise. An alternative method to identify sugarcane borers is the use of molecular data. DNA barcoding based on partial cytochrome c oxidase subunit 1 (COI) sequences has proven to be a useful tool for rapid and accurate species determination in many insect taxa. This study was conducted to test the effectiveness of DNA barcodes to discriminate among sugarcane borer species in China. Partial sequences of the COI gene (709 bp) were obtained from six species collected from different geographic areas. Results showed that the pairwise intraspecies genetic distance was < 0.02, whereas the interspecies genetic distance ranged from 0.117 to 0.182. Results from a neighbor-joining tree showed that the six sugarcane borer species were certainly separated. These results suggested that the partial COI sequences had high barcoding resolution in discriminating among sugarcane borer species. Our study emphasized the use of DNA barcodes for identification of the analyzed sugarcane borer species and represents an important step for building a comprehensive barcode library for sugarcane borers in China.


T-cell functional defects in rheumatoid arthritis: intrinsic or extrinsic?
This study investigated two mechanisms which may underlie abnormal T-cell function [lymphocyte proliferation and interleukin-2 (IL-2) production] in rheumatoid arthritis (RA). These were: (a) a possible lack of the IL-2-producing CD4+2H4+ lymphocytes and (b) the possible inhibitory role of monocytes and neutrophils. Numbers of CD4+2H4+ cells did not differ between normal controls and patients with RA, although IL-2 produced by the peripheral blood mononuclear cells (PBMC) of the same individuals was markedly reduced in the patient group (P less than 0.001). Many rheumatoid peripheral blood mononuclear cell preparations, but very few control, were contaminated with neutrophils (P less than 0.001). This was more marked in patients with active RA than in those with inactive disease (P less than 0.001). Numbers of monocytes were similar in all groups. Monocyte depletion, or addition of indomethacin and/or catalase in PBMC, caused a significantly greater increase of responses in RA patients than in controls. This effect was significantly higher in patients with active disease than in the inactive group. These findings suggest that activated monocytes and neutrophils found in the rheumatoid PBMC preparations exert inhibitory effects mediated, in part, by the production of prostaglandins and reactive oxygen intermediates. Monocyte depletion and partial reconstitution resulted in significant increase of lymphocyte proliferation and IL-2 production in both controls and patients. None of the manipulations performed succeeded in normalizing the deficient rheumatoid T-cell responses. These data support the hypothesis that non-lymphoid cell populations play an important role in the T-cell dysfunction characteristic of RA.(ABSTRACT TRUNCATED AT 250 WORDS)


The mitigating effect of Spirulina (Arthrospira platensis) on the hemotoxicity of gibberellic acid on juvenile tilapia (Oreochromis niloticus).
The use of plant growth regulators has led to environmental contamination of water bodies that occur adjacent to agricultural areas. Some of these chemicals are bioactive, not only to plants, but also to non-target exposed biota, namely of the aquatic compartment. Previous work demonstrated the establishment of hepato- and nephrotoxic effects in juvenile tilapia (Oreochromis niloticus) exposed via aquatic media to gibberellic acid (GA3), which is among the most used plant growth regulators, in agricultural practices. Here, we investigated the effect of GA3 on hematological indices, poikilocytosis, nuclear abnormalities, and genotoxic indices measured in Nile tilapia (Oreochromis niloticus), as well as the putative protective effects of dietary supplementation of Spirulina (Arthrospira platensis). Fish were evenly assorted into 5 groups: group I served as a control, and groups II-V were fed diets supplemented with Spirulina at rates of 0 g/kg, 5 g/kg, 20 g/kg, and 100 g/kg, respectively, for 2 months before being exposed to 150 mg/L GA3. The results revealed that GA3 exposure decreased significantly all hematological indices (P < 0.05), except leucocytes and mean corpuscular hemoglobin concentration (MCHC), compared to the control group (P > 0.05). GA3 exposure increased significantly the percentage of nuclear abnormalities, altered erythrocytes and the percentages of tail DNA, compared to the control group (P < 0.05). Spirulina supplementation restored the hematological, poikilocytosis, nuclear abnormalities, and the percentages of tail DNA to near normal levels. The 100 g/kg SP treatment was the most effective in attaining such effect, showing concentration-dependency. The present study reinforces our findings of the toxicity of GA3 on O. niloticus and suggests that the addition of Spirulina to fish diet can mitigate the hemotoxic effects of GA3.


Toxic epidermal necrolysis secondary to carbamazepine in a patient with human immunodeficiency virus infection.
Toxic epidermal necrolysis (TEN) is a rare but very serious dermatologic disorder and is seen more commonly in human immunodeficiency virus (HIV) infected patients. We present a case of TEN in HIV infected person secondary to carbamazepine who responded favourably to corticosteroids.


Combined and alternating ganciclovir and foscarnet in acute and maintenance therapy of human immunodeficiency virus-related cytomegalovirus encephalitis refractory to ganciclovir alone. A case report and review of the literature.
Cytomegalovirus (CMV) causes life-threatening disseminated infections and in particular vision-threatening infections of the retina in patients with the acquired immunodeficiency syndrome. Ganciclovir currently represents the most frequently used therapy for CMV retinitis. However, cases of ganciclovir-resistant CMV strains have been described, in which foscarnet seems to be an effective alternative. Both drugs have serious toxicities, and relapses frequently occur during maintenance therapy. In a patient with CMV encephalitis, we administered a 3-week combination ganciclovir/foscarnet induction therapy (ganciclovir 5 mg/kg every 12 h; foscarnet 60 mg/kg every 8 h), followed by an alternating maintenance administration of both drugs every other day (ganciclovir 5 mg/kg, foscarnet 120 mg/kg) to reduce toxicity and resistance. This regimen was tolerated well and seemed to be more effective than ganciclovir alone in a patient with CMV encephalitis.


CGE modeling with disaggregated pollution treatment sectors for assessing China's environmental tax policies.
This research involved constructing a computable general equilibrium (CGE) model for assessing China's latest environmental tax policies. Most environmental CGE models link pollutant emissions to the standard CGE model only by pollution coefficients per unit of sectoral output, and the emission reduction process is not included within production structures. We constructed separate pollution treatment sectors for solid waste management, wastewater management, and waste gas management to describe the pollution treatment processes and identify how policies affect production activities. We compiled the satellite accounts of 18 pollutants from the China Environmentally Extended Input-Output (CEEIO) dataset covering primary gas, water, and solid pollutants and disaggregated the electricity sector into six different production technologies: hydroelectricity, coal power, gas electricity, oil electricity, nuclear power, and renewable energies. We drew two primary conclusions from the simulation results. First, the environmental policies examined could help reduce the emissions of most kinds of pollutants, but also negatively affect GDP. GDP loss by 2030 would be 0.03% in the low environmental tax scenario (LowET), 0.06% in the high environmental tax scenario (HighET), 0.16% in the low environmental tax and low carbon tax scenario (LowETC), and 0.34% in the high environmental tax and high carbon tax scenario (HighETC). SO2 emissions would decrease by 17.4%, 21.0%, 19.3% and 24.5%, respectively, and CO2 emissions would reduce by 0.9%, 1.7%, 5.8% and 11.0%. Second, despite the minor changes in the economic impacts, the effectiveness in pollution treatment of environmental tax policies is underestimated if the pollution treatment sectors are disaggregated in the CGE model. Take the SO2 for an example. The calculated SO2 reductions will increase from 8.95% to 24.46% after disaggregating the pollution treatment sectors in HighETC scenarios.


Is anthelmintic resistance a concern for heartworm control? What can we learn from the human filariasis control programs?
Heartworm prophylaxis is currently largely dependent on the ability of avermectins and milbemycins to arrest the development of third and fourth stages of Dirofilaria immitis for prolonged periods, without producing adulticidal effects. Major control programs, dependent on the activity of ivermectin, are being implemented for human onchocerciasis and lymphatic filariasis. The avermectins and milbemycins act on glutamate-gated and gamma-aminobutyrate-gated chloride channel subunit proteins in nematodes. Ivermectin resistance has been widely described in trichostrongylid nematodes of ruminants. There is evidence that when ivermectin resistance occurs in nematodes, there may be selection on some, but not all of the genes that code for ligand-gated chloride channel subunit proteins as well as on some ABC-transporter genes, whose products may be involved in regulating macrocyclic lactone drug concentrations at receptors, and on some structural protein genes of amphidial neurones. Although ivermectin resistance has not been reported in filarial nematodes, there have recently been reports of suboptimal responses to ivermectin in Onchocerca volvulus. Evidence has been found of ivermectin selection on at least ABC-transporter genes and some neuronal structural protein genes in O. volvulus. To date, there is no evidence of avermectin/milbemycin resistance in D. immitis, also a filarial nematode. Chemotherapy against trichostrongylids of animals, human filariae, and D. immitis, relies on avermectins or milbemycins. However, control involves targeting different stages or processes in the nematode life cycles, different control strategies, different proportions of the nematode population in refugia, and different drug dosage rates. Consideration of the proportion of the D. immitis population normally in refugia, the life cycle stage targeted, and the anthelmintic dosages used suggest that it is unlikely that significant avermectin/milbemycin resistance will be selected in D. immitis with current treatment strategies.


Perineural invasion by squamous-cell carcinoma of the lower lip. Review of the literature and report of a case.
Although perineural metastases have been studied extensively for various regions of the body, there is a relative paucity of data on cancers of the head and neck region. A case of perineural metastasis from a squamous-cell carcinoma of the lower lip is presented, with symptoms mimicking other neurologic diseases.


Genomic, probiotic, and metabolic potentials of Liquorilactobacillus nagelii AGA58, a novel bacteriocinogenic motile strain isolated from lactic acid-fermented shalgam.
This study aimed to perform genomic, probiotic, and metabolic characterization of a novel Liquorilactobacillus nagelii AGA58 isolated from a lactic acid-fermented shalgam beverage to understand its metabolic potentials and probiotic features. AGA58 is gram-positive, motile, catalase-negative and appears as short rods under the light-microscope. The AGA58 chromosome comprises a single linear chromosome of 2,294,635 bp that is predicted to carry 2135 coding sequences, including 45 tRNA genes, 3 mRNA, and 3 rRNA operons. The genome has a G+C content of 36.9%, including 55 pseudogenes and a single intact prophage. AGA58 is micro-anaerobic due to achieving a shorter doubling time and faster growth rate than micro-aerophilic conditions. It carries flagellar biosynthesis protein-encoding genes predicting motile behavior, which was confirmed with the in vitro motility test. AGA58 is an obligatory homofermentative lactobacillus that can ferment hexose sugars such as galactose, glucose, fructose, sucrose, mannose, N-acetyl glucosamine, maltose, and trehalose to lactate through glycolysis. No acid production from pentoses implies that five-carbon sugars are being utilized for purine and pyrimidine synthesis. Putative pyruvate metabolism revealed formate, malate, oxaloacetate, acetate, acetaldehyde, acetoin, and lactate forms from pyruvate. AGA58 is predicted to encode the LuxS gene and biosynthesis of class IIa and Blp family class-II bacteriocins suggesting this bacterium's antimicrobial potential, linked to antagonism tests that AGA58 can inhibit Escherichia coli ATCC 43895, Salmonellaenterica serovar Typhimurium ATCC 14028, and Klebsiellapneumonia ATCC 13883. Moreover, AGA58 is tolerant to acid and bile concentrations simulating the human gastrointestinal conditions depicting the probiotic potential of the organism as the first report in literature within the same species.


Hepatitis C virus vaccines--progress and perspectives.
Approximately 170 million individuals, representing 3% of the global population, are infected with hepatitis C virus (HCV). Whereas strategies for antiviral therapies have markedly improved resulting in clinical licensing of direct-acting antivirals, the development of vaccines has been hampered by the high genetic variability of the virus as well as by the lack of suitable animal models for proof-of-concept studies. Nevertheless, there are several promising vaccine candidates in preclinical and clinical development. After a brief summary of the molecular virology and immunology relevant to vaccine development, this review explains the model systems used for preclinical vaccine development, and highlights examples for most recently developed HCV vaccine candidates.


Methodologic issues in the measurement of interleukin-16 in clinical blood samples using immunoassays.
Quantitation of interleukin-16 (IL-16) in clinical blood samples has strongly increased, since IL-16 appears to be involved in the pathogenesis of several inflammatory diseases. IL-16 is synthesized in the cell cytoplasm as precursor protein (pro-IL-16), which can be processed by caspase-3 into N-terminal (N-IL-16) and C-terminal (C-IL-16) fragments. C-IL-16 is described to be subsequently secreted. Using commercially available IL-16 ELISA, a pro-IL-16 ELISA and immunoprecipitation analysis, we investigated, whether type and handling of blood samples influence IL-16 quantitation and whether existing IL-16 ELISA are specific for C-IL-16. We observed that cell-rich plasma samples reflect falsely-elevated IL-16 concentrations due to cell contaminations. Interestingly, not C-IL-16, but pro-IL-16 represents the major IL-16 form in cell-rich plasma samples. Notably, commercially IL-16 ELISA could not distinguish between C-IL-16 and pro-IL-16. Thus, cell-rich plasma samples should not be used for IL-16 measurements and new methods are necessary for quantitation of C-IL-16 and pro-IL-16 uniquely.


An audit of neonatal and infant hepatitis B immunisation and serological testing in two counties of England, 2007-12.
We audited adherence to national hepatitis B virus (HBV) immunisation policy for neonates and infants born to HBV positive mothers in two counties of England during 2007/08 to 2011/12 (n=112 in County X, n=190 in County Y). Over the five year period, 29.9% of at risk neonates in County X and 23.5% in County Y required hepatitis B immunoglobulin (HBIG) at birth. The annual median age of HBIG administration was 0.0-0.5 days. The annual median coverage and timeliness of the first (coverage range 92.3-100.0%; age of administration range 0.0-0.0 days), second (83.8-100.0%; 32.0-42.0 days), third (81.1-100.0%; 62.0-81.0 days) and fourth dose HBV immunisations (44.4-91.9%; 378.0-443.0 days) and serological testing (8.6-81.0%; 450.0-707.0 days) were calculated. Statistically significant variation was found in the coverage of third and fourth dose immunisations in County Y, age of fourth dose immunisation in County X, and the coverage and timeliness of serological testing in both counties (p < 0.05). HBIG and the first three HBV immunisations were commonly administered according to the national schedule. Fourth dose immunisations and serological tests showed poor adherence. We advocate public health interventions to improve immunisation programme outcomes and hepatitis B surface antigen testing.


Polymorphic variants of drug-metabolizing enzymes alter the risk and survival of oral cancer patients.
The present study investigated the prevalence of CYP2D6*4, CYP3A5*3 and SULT1A1*2, using PCR-RFLP, in normal and oral cancer (OC) patients that were stratified by OC subtype and gender. The risk of cancer, 5-year cumulative survival and hazard's ratio (HR) with respect to risk factors and clinical factors were estimated using Fisher's exact test, Kaplan-Meier analysis, and Cox proportional hazards models. CYP2D6*4 'GA' lowered the risk of buccal mucosa cancer (BMC) in males (OR = 0.37), whereas, 'G' allele of CYP3A5*3 increased risk of tongue cancer (TC) (OR = 1.67). SULT1A1*2 'GA' increased the risk of TC (OR = 2.36) and BMC (OR = 3.25) in females. The 5-year survival of the patients depended on factors like age, lymphovascular spread (LVS), perinodal spread (PNS), recurrence, tobacco, and alcohol. CYP3A5*3 'AG' and 'GG' had decreased the hazard ratio (HR) for BMC females when inflammatory infiltrate alone or along with other covariates, LVS, PNI, PNS, metastasis, recurrence, and relapse was adjusted. Similarly, CYP3A5*3 'AG' decreased the risk of death (HR = 0.05) when the grade was adjusted. SULT1A1*2 'GA' had decreased HR for TC males (HR = 0.08) after adjusting for inflammatory infiltrate, LVS, perineural invasion (PNI), PNS, metastasis, recurrence, and relapse. Further, our bioinformatics study revealed the presence of a CpG island within the CYP2D6 and a CTCF binding site upstream of CYP2D6. Interestingly, three CpG islands and two CTCF binding sites were also identified near the SULT1A1. In conclusion, the SNPs altered risk and survival of BMC and TC differentially in a gender specified manner, that varied with clinical and risk factors.


Mouthing of Soil Contaminated Objects is Associated with Environmental Enteropathy in Young Children.
To characterise childhood mouthing behaviours and to investigate the association between object-to-mouth and food-to-mouth contacts, diarrhoea prevalence and environmental enteropathy. A prospective cohort study was conducted of 216 children ≤30 months of age in rural Bangladesh. Mouthing contacts with soil and food and objects with visible soil were assessed by 5-h structured observation. Stool was analysed for four faecal markers of intestinal inflammation: alpha-1-antitrypsin, myeloperoxidase, neopterin and calprotectin. Overall 82% of children were observed mouthing soil, objects with visible soil, or food with visible soil during the structured observation period. Sixty two percent of children were observed mouthing objects with visible soil, 63% were observed mouthing food with visible soil, and 18% were observed mouthing soil only. Children observed mouthing objects with visible soil had significantly elevated faecal calprotectin concentrations (206.81 μg/g, 95% confidence interval [CI]: 6.27, 407.36). There was also a marginally significant association between Escherichia coli counts in soil from a child's play space and the prevalence rate of diarrhoea (diarrhoea prevalence ratio: 2.03, 95% CI 0.97, 4.25). These findings provide further evidence to support the hypothesis that childhood mouthing behaviour in environments with faecal contamination can lead to environmental enteropathy in susceptible paediatric populations. Furthermore, these findings suggest that young children mouthing objects with soil, which occurred more frequently than soil directly (60% vs. 18%), was an important exposure route to faecal pathogens and a risk factor for environmental enteropathy.


V1 neurons: in tune with the neighbors.
In this issue of Neuron, Nauhaus et al. use a combination of optical imaging and multiple electrode recording to demonstrate that the orientation tuning of single cells in primary visual cortex is reliably related to the local structure of the orientation preference map in both cats and monkeys.


Adsorption of doxorubicin hydrochloride on glutaric anhydride functionalized Fe3O4@SiO2 magnetic nanoparticles.
Since Fe3O4 nanoparticles synthesized by plant extracts possess good bio-compatibility and superparamagnetic properties, the possibility of these could be used as a carrier in drug delivery. In this work, doxorubicin hydrochloride (DOX), an anti-cancer drug, loaded on glutaric anhydride-functionalized magnetic nanoparticles (Fe3O4@SiO2-Glu) was investigated at varying pH values for effective drug delivery. Various factors affecting the adsorption of DOX onto the Fe3O4@SiO2-Glu were examined, where the adsorption efficiency of DOX reached 92% at a concentration of 20 mg/L employing 10 mg of Fe3O4@SiO2-Glu at 303 K in pH 7.4. However, the adsorption efficiency of DOX was decreased to 18% at acidic pH value down to 3.0, implicating that the drug releasing process was controlled by pH. Adsorption kinetics was fitting to pseudo-second-order and the isothermal adsorption conformed to Freundlich isotherm. The morphology and surface composition of the synthesized Fe3O4@SiO2-Glu were characterized by SEM, TEM, and N2 adsorption/desorption isotherms, revealing that the specific surface area being 62.6 m2/g and the size ranging from ~30 to 50 nm. The zeta potential results indicated that Fe3O4@SiO2-Glu were negatively charged in various pH from 3 to 8.5. Characterizations by FTIR and UV-Vis techniques suggested that the DOX was absorbed and it can be delivered by Fe3O4@SiO2-Glu.


Clinical characteristics and STK11 gene mutations in Chinese children with Peutz-Jeghers syndrome.
Peutz-Jeghers syndrome (PJS) is a rare autosomal dominant inherited disease characterized by gastrointestinal hamartomatous polyps and mucocutaneous melanin spots. Germline mutation of the serine/threonine kinase 11 (STK11) gene are responsible for PJS. In this study, we investigated the clinical characteristics and molecular basis of the disease in Chinese children with PJS. Thirteen children diagnosed with PJS in our hospital were enrolled in this study from 2011 to 2015, and their clinical data on polyp characteristics, intussusceptions events, family histories, etc. were described. Genomic DNA was extracted from whole-blood samples from each subject, and the entire coding sequence of the STK11 gene was amplified by polymerase chain reaction and analyzed by direct sequencing. The median age at the onset of symptoms was 2 years and 4 months. To date, these children have undergone 40 endoscopy screenings, 17 laparotomies and 9 intussusceptions. Polyps were found in the stomach, duodenum, small bowel, colon and rectum, with large polyps found in 7 children. Mutations were found in eleven children, including seven novel mutations (c.481het_dupA, c.943_944het_delCCinsG, c.397het_delG, c.862 + 1G > G/A, c.348_349het_delGT, and c.803_804het_delGGinsC and c.121_139de l19insTT) and four previously reported mutations (c.658C > C/T, c.890G > G/A, c.1062 C > C/G, and c.290 + 1G > G/A). One PJS patient did not have any STK11 mutations. The polyps caused significant clinical consequences in children with PJS, and mutations of the STK11 gene are generally the cause of PJS in Chinese children. This study expands the spectrum of known STK11 gene mutations.


Reference values for serum cystatin C in very low-birthweight infants: From two centres of China.
To determine the level of cystatin C (Cys-C) values in preterm babies for the purpose of becoming a good endogenous marker of renal function. A total of 366 very low-birthweight infants (including 70 extremely low-birthweight babies) with gestational age <37 weeks born in two centres were studied. In very low-birthweight infants, the mean level of Cys-C was 1.96 ± 0.44 mg/L in blood samples taken on day 1, 1.78 ± 0.49 mg/L on day 7 and 1.71 ± 0.47 mg/L on day 28. In extremely low-birthweight infants, the mean level of Cys-C was 2.00 ± 0.49 mg/L on day 1, 1.63 ± 0.38 mg/L on day 7 and 1.62 ± 0.55 mg/L on day 28, respectively. Compared to serum creatinine and blood urea nitrogen, Cys-C level was independent of birthweight and gestational age. Cys-C is regarded as an alternative for assessing renal function in very low-birthweight infants, but its advantages over serum creatinine and blood urea nitrogen has not been fully proved yet. Hence, larger sample study is still necessary.


The effect of hemolysis on quality control metrics for noninvasive prenatal testing.
Noninvasive prenatal testing (NIPT) is the testing of blood samples from pregnant women to screen for fetal risk of chromosomal disorders. Even though in vitro hemolysis of blood specimens is common in clinical laboratories, its influence on NIPT has not been well investigated. Peripheral blood samples were collected from 205 pregnant women and categorized according to the concentration of free hemoglobin in the plasma. After performing NIPT using massively parallel sequencing, the quality control metrics were analyzed and compared with samples that did not undergo hemolysis or samples redrawn from the same women. The specimens were divided into four groups based on the concentration of free hemoglobin: Group I (0-1 g/L, n = 53), Group II (1-2 g/L, n = 97), Group III (2-4 g/L, n = 30), and Group IV (> 4 g/L, n = 25). There was no significant difference in the quality control metrics of clinical samples with slight or moderate hemolysis (Group II and III). However, samples with severe hemolysis (Group IV) showed a significantly increased rate of duplicated reads (duplication rate) and fetal fraction, as well as decreased library concentration compared with samples without hemolysis. Moreover, the increase in fetal fraction caused by hemolysis was confirmed by redrawing blood samples in Group IV. For NIPT using massively parallel sequencing, samples with slight or moderate hemolysis (≤ 4 g/L) are acceptable. However, careful consideration should be taken regarding the use of severely hemolyzed samples (> 4 g/L), since they might increase the risk of test failure.


Correction: Study on the transcriptional regulatory mechanism of the MyoD1 gene in Guanling bovine.
[This corrects the article DOI: 10.1039/C7RA11795G.].


Isolated lesion of the medial orbital wall following endonasal surgery. Isolated fractures of the medial orbital wall.
Isolated fractures of the medial orbital wall are infrequent. The diagnostic triad includes: adduction block, exotropia with diplopia in all directions of gaze, positive passive duction in abduction. Sometimes a slight enophthalmos is present. Computed tomography shows the extension and the seat of the fracture. The authors illustrate the case of a 60 year old male who presented with a breach of the medial orbital wall following endonasal surgery. The patient was successfully operated using an iliac bone graft inserted via an eyebrow-nasal cutaneous approach, after a previous attempt with a transconjunctival approach performed in another hospital had failed. A good functional and aesthetic result was observed within the first year after surgery. After almost 11 years a full adduction is still present and diplopia is absent. The authors underline the importance of an early diagnosis and prompt surgical treatment. The fat-muscle entrapment should be removed and the bone defect closed. A close cooperation between ophthalmologist and plastic surgeon is suggested.


Type C Niemann-Pick disease. Lysosomal accumulation and defective intracellular mobilization of low density lipoprotein cholesterol.
The intracellular accumulation of unesterified cholesterol was examined during 24 h of low density lipoprotein (LDL) uptake in normal and Niemann-Pick C fibroblasts by fluorescence microscopy with filipin staining and immunocytochemistry. Perinuclear fluorescence derived from filipin-sterol complexes was observed in both normal and mutant cells by 2 h. This perinuclear cholesterol staining reached its peak in normal cells at 6 h. Subsequent development of fluorescence during the remaining 18 h of LDL incubation was primarily limited to the plasma membrane region of normal cells. In contrast, mutant cells developed a much more intense perinuclear fluorescence throughout the entire 24 h of LDL uptake with little enhancement of cholesterol fluorescence staining in the plasma membranes. Direct mass measurements confirmed that internalized LDL cholesterol more readily replenishes the plasma membrane cholesterol of normal than of mutant fibroblasts. Perinuclear filipin-cholesterol fluorescence of both normal and mutant cells was colocalized with lysosomes by indirect immunocytochemical staining of lysosomal membrane protein. Abnormal sequestration of LDL cholesterol in mutant cells within a metabolically latent pool is supported by the finding that in vitro esterification of cellular cholesterol could be stimulated in mutant but not in normal cell homogenates by extensive disruption of the intracellular membranous structures of cells previously cultured with LDL. Deficient translocation of exogenously derived cholesterol from lysosomes to other intracellular membrane sites may be responsible for the delayed homeostatic responses associated with LDL uptake by mutant Niemann-Pick Type C fibroblasts.