How to Make a Fast, Efficient Bubble-Driven Micromotor: A Mechanical View.
Micromotors, which can be moved at a micron scale, have special functions and can perform microscopic tasks. They have a wide range of applications in various fields with the advantages of small size and high efficiency. Both high speed and efficiency for micromotors are required in various conditions. However, the dynamical mechanism of bubble-driven micromotors movement is not clear, owing to various factors affecting the movement of micromotors. This paper reviews various factors acting on micromotor movement, and summarizes appropriate methods to improve the velocity and efficiency of bubble-driven micromotors, from a mechanical view. The dynamical factors that have significant influence on the hydrodynamic performance of micromotors could be divided into two categories: environment and geometry. Improving environment temperature and decreasing viscosity of fluid accelerate the velocity of motors. Under certain conditions, raising the concentration of hydrogen peroxide is applied. However, a high concentration of hydrogen peroxide is not applicable. In the environment of low concentration, changing the geometry of micromotors is an effective mean to improve the velocity of micromotors. Increasing semi-cone angle and reducing the ratio of length to radius for tubular and rod micromotors are propitious to increase the speed of micromotors. For Janus micromotors, reducing the mass by changing the shape into capsule and shell, and increasing the surface roughness, is applied. This review could provide references for improving the velocity and efficiency of micromotors.


Validity of self-rated health among Latino(a)s.
The authors investigated whether self-rated health (SRH) had differential mortality risks for Latino(a) adults of various acculturation statuses living in the United States. They used cumulative National Health Interview Survey data from 1989 to 1994 (n = 37,713) linked with the National Health Interview Survey Multiple Cause of Death data files (1,364 deaths) that match records from the National Death Index through 1997. The authors specified survival models to estimate the effect of SRH on mortality and further stratified their model by birth and duration in the United States as proxies for acculturation. These estimates were compared across strata. Poor SRH was found to be a weaker predictor of subsequent mortality risk among the less acculturated, although the overall risk among the aggregated sample is similar to the risk reported in previous studies. The relation between poor SRH and mortality risk increases with United States acculturation among Latinos. While poor SRH was significantly associated with short-term mortality among the least acculturated, this association did not persist beyond 2-year mortality risk. Health researchers wishing to use SRH to assess the physical health of multiethnic populations should at least control for levels of acculturation among respondents.


The effect of Kinesio® taping on pain, functionality, mobility and endurance in the treatment of chronic low back pain: A randomized controlled study.
Low back pain is one of the most important causes of morbidity. This study was designed to evaluate the effect of Kinesio® taping on pain, functionality, mobility and endurance in chronic low back pain treatment. Patients with chronic low back pain were randomly divided into three groups. Therapeutic ultrasound, hot packs, and transcutaneous electrical nerve stimulation were applied to each group for ten sessions during two weeks, and therapeutic exercises were applied in the clinic under physiotherapist supervision starting from the sixth session. Kinesio® tape was applied to the patients in the first group after each treatment session, and placebo tape was applied to the patients in the second group. No taping was applied to the third group, which constituted the control group. All the patients were evaluated pre and post-treatment in respect of pain, functional status (Oswestry scale), flexibility and endurance. The study included 60 patients (32 females). When the initial demographic and clinical characteristics of the groups were evaluated, all assessment results, except the Oswestry scores, were similar (p= 0.000). When the average changes in the clinical evaluations were evaluated after the treatment, a statistically significant improvement demonstrating the superiority of the taping group was observed in pain, functionality, flexibility and endurance values (p= 0.000, 0.000, 0.000, 0.000). Kinesio® taping in chronic low back pain is an easy and effective method which increases the effectiveness of the treatment significantly in a short period when applied in addition to exercise and electrotherapy methods.


Chapter 10: Sinonasal malignancies.
Malignant tumors of the sinonasal tract are uncommon tumors of the head and neck. Patients often present in the later years of life with unilateral symptoms and potential involvement of nearby structures such as the orbit, brain, or cranial nerves. Presenting symptoms are similar to patients suffering from inflammatory sinonasal disease and thus early diagnosis relies heavily on a high clinical suspicion. There are established risk factors based on exposure to the by-products of woodworking, metal, textile, and leather industries. Sinonasal malignancies are generally divided into those of epithelial origin (squamous cell carcinoma, adenocarcinoma, and adenoid cystic carcinoma) and nonepithelial origin (olfactory neuroblastoma, chondrosarcoma, and mucosal melanoma). Accurate histopathology confirmation and staging of the tumor is critical prior to making treatment decisions. Both computed tomography and magnetic resonance imaging are required to accurately determine the extent of local disease. Treatment is based on multimodality therapy, primarily surgical excision, and postoperative radiotherapy. This article reviews the classification of malignant tumors of the paranasal sinuses, their clinical presentation, relevant diagnostic investigations, and the principals of therapy and management.


Maternal separation changes maternal care, anxiety-like behaviour and expression of paraventricular oxytocin and corticotrophin-releasing factor immunoreactivity in lactating rats.
The early postnatal period is a time of tremendous change for the dam and her offspring. During this time, environmental insults such as repeated stress exposure can have detrimental effects. In research that has focused on the effect of postnatal stress exposure on the dams, conflicting changes in maternal care and anxiety-like behaviour have been reported. Additionally, changes to hypothalamic neuropeptides that are crucially involved in the transition to motherhood and stress regulation, namely oxytocin and corticotrophin-releasing factor (CRF), have not been examined. Accordingly, the present study aimed to determine (i) whether repeated postpartum stress increases engagement in maternal care behaviours and anxiety-like behaviour and (ii) whether these behavioural changes correspond with changes to CRF- or oxytocin-immunoreactive (-IR) cells in the paraventricular nucleus (PVN) of the hypothalamus. A non-lactating group was also included to control for the effects of lactation on anxiety and the hypothalamic neuroendocrine system. Following the birth of their litters, Long-Evans dams were separated from their pups from postnatal day (PND) 1 to PND21 for either 15 minutes (maternal separation [MS]15) or 6 hours (MS360). Maternal behaviours were recorded for 30 minutes on select PNDs following the separation. On PND22, dams were exposed to the elevated plus maze, brains were collected, and immunofluorescence analysis of PVN oxytocin- and CRF-IR cells was conducted. Our findings demonstrate that prolonged maternal separation altered typical maternal behaviours and reduced anxiety relative to MS15 dams. At the cellular level, oxytocin-IR cells in the caudal PVN were reduced in MS360 dams to a level similar to that in non-lactating controls, and PVN CRF-IR cells were reduced relative to both MS15 and non-lactating controls. Taken together, these data reveal the behavioural and neuronal changes that occur in the mother dam following repeated postnatal stress exposure.


Pharmacokinetic/Pharmacodynamic Profiles of Tiamulin in an Experimental Intratracheal Infection Model of Mycoplasma gallisepticum.
Mycoplasma gallisepticum is the most important pathogen in poultry among four pathogenic Mycoplasma species. Tiamulin is a pleuromutilin antibiotic that shows a great activity against M. gallisepticum and has been approved for use in veterinary medicine particularly for poultry. However, the pharmacokinetic/pharmacodynamics (PK/PD) profiles of tiamulin against M. gallisepticum are not well understood. Therefore, in the current studies, we investigated the in vivo PK/PD profiles of tiamulin using a well-established experimental intratracheal infection model of M. gallisepticum. The efficacy of tiamulin against M. gallisepticum was studied in 8-day-old chickens after intramuscular (i.m.) administration at 10 doses between 0-80 mg/kg. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to evaluate the PK parameters of tiamulin following i.m. administration at doses of 5, 40, and 80 mg/kg in Mycoplasma gallisepticum-infected neutropenic chickens. Real-time PCR (RT-PCR) was used for quantitative detection of M. gallisepticum. The MIC of tiamulin against M. gallisepticum strain S6 was 0.03 μg/mL. The PK/PD index, AUC24h/MIC, correlated well with the in vivo antibacterial efficacy. The in vivo data suggest that animal dosage regimens should supply AUC24h/MIC of tiamulin of 382.68 h for 2 log10 ccu equivalents M. gallisepticum reduction. To attain that goal, the administered dose is expected to be 45 mg/kg b.w. for treatment of M. gallisepticum infection with an MIC90 of 0.03 μg/mL.


Simultaneous carotid endarterectomy and coronary artery bypass surgery in Canada.
Stroke neurologists are commonly asked to review patients who require coronary artery bypass grafting (CABG) but who also have comorbid severe carotid stenosis; such patients may be offered simultaneous carotid endarterectomy (CEA). In Canada, 0.51% of CABG procedures were combined CEA-CABG. The adjusted stroke and death rate was 2.67-fold greater in the combined CEA-CABG group compared to CABG alone. Randomized trials of the combined procedure are needed.


Physical conditioning and mental stress reduction--a randomised trial in patients undergoing cardiac surgery.
Preoperative anxiety and physical unfitness have been shown to have adverse effects on recovery from cardiac surgery. This study involving cardiac surgery patients was primarily aimed at assessing the feasibility of delivering physical conditioning and stress reduction programs within the public hospital setting. Secondary aims were to evaluate the effect of these programs on quality of life (QOL), rates of postoperative atrial fibrillation (AF) and length of stay (LOS) in hospital. Elective patients scheduled for coronary artery bypass graft and/or valve surgery at a public hospital in Melbourne, Australia were enrolled. Patients were randomized to receive either holistic therapy (HT) or usual care (UC). HT consisted of a series of light physical exercise sessions together with a mental stress reduction program administered in an outpatient setting for the first two weeks after placement on the waiting list for surgery. A self-administered SF-36 questionnaire was used to measure QOL and hospital records to collect data on LOS and rate of postoperative AF. The study population comprised 117 patients of whom 60 received HT and 57 received UC. Both programs were able to be delivered within the hospital setting but ongoing therapy beyond the two week duration of the program was not carried out due to long waiting periods and insufficient resources. HT, as delivered in this study, compared to UC did not result in significant changes in QOL, LOS or AF incidence. Preoperative holistic therapy can be delivered in the hospital setting, although two weeks is insufficient to provide benefits beyond usual care on QOL, LOS or postoperative AF. Further research is now required to determine whether a similar program of longer duration, or targeted to high risk patients can provide measurable benefits. This trial was conducted as part of a larger study and according to the principles contained in the CONSORT statement 2001.


Extracellular matrix gene regulation.
Extracellular matrix metabolism plays a central role in development of skeletal tissues and in most orthopaedic diseases and trauma such as fracture or osteotomy repair, arthritis, cartilage repair, and congenital skeletal deformity. During development or disease, specific genes must be expressed in order to make or repair appropriate extracellular matrix. For example, specific gene expression patterns are characteristic of bone and cartilage. The precise expression pattern depends on a balance of positive and negative transcription factors, proteins that control the synthesis of mRNA from the specific gene. In cartilage, a number of studies indicate that Sox transcription factors are critical positive regulators in genes such as COL2A1, COL9A2, COL11A2, aggrecan, and CD-RAP. In addition, negative regulators are also essential to fine tune gene regulation in chondrocytes and to turn off gene expression in noncartilaginous tissues. Negative transcription factors in cartilage include partial differentialEF-1, snail/slug, CYRBP1, NT2, and C/EBP. Runx2 and osterix are critical transcription factors for osteogenesis but also have some influence on chondrogenesis. The availability of cis-regulatory sites in specific genes combined with the availability of transcription factors in the nucleus determines the level of gene expression.


Theoretical and experimental study of the bleaching of a dye in a film-polymerization process.
The quantum efficiency and the molar-absorption coefficients of different phenothiazine dyes are obtained by means of fitting the experimental data of transmittance as a function of time. An analytical expression for the intensity transmitted in a photopolymerizable holographic material is obtained, and good agreement between theory and experience is also achieved. The analysis of these parameters is of fundamental quantities in the photochemical characterization of holographic recording materials.


Fiberoptic intraparenchymal brain pressure monitoring with the Camino V420 monitor: reflections on our experience in 163 severely head-injured patients.
To assess the safety and accuracy of the Camino intraparenchymal sensor, we prospectively evaluated hemorrhagic complications, zero-drift, infection, and system malfunction in 163 patients monitored after a severe head injury. Mean duration of intracranial pressure (ICP) monitoring was 5 +/- 2.2 days (range: 12 h to 11 days). Of the 141 patients with a control CT scan, four showed a 1-2-cc collection of blood at the catheter's end. When removed, the sensors underread the true ICP value (negative zero-drift) in 80 of the 126 sensors evaluated (63.5%). Fourteen sensors showed no zero-drift, and 32 sensors overread the true ICP value (positive zero-drift) (median: -1 mm Hg; interquartile range: -4 to +1 mm Hg). No significant relationship was found between zero-drift, the surgeon who implanted the sensor, intracranial hypertension, or duration of ICP monitoring. No clinical infections could be attributed to the devices. Sixteen patients (9.8%) required more than one ICP sensor due to malfunctioning of the system. In conclusion, continuous ICP monitoring using the Camino intraparenchymal sensor has a low complication rate. However, this sensor may underread the real ICP values in a high number of patients. The lack of correlation between duration of ICP monitoring and zero-drift suggests that, contrary to the recommendations of other reports, the intraparenchymatous Camino sensor can provide reliable readings after the fifth day of use.


Cissus quadrangularis (veldt grape) attenuates disease progression and anatomical changes in mono sodium iodoacetate (MIA)-induced knee osteoarthritis in the rat model.
The Cissus quadrangularis (CQ) stem has interesting nutritional and pharmacological properties to promote the health of the skeletal system. It is a well-recognized plant in the conventional system of medicine in India for treating bone and joint-associated complications. This study focuses on identifying the active constituents from the stem and root extracts of CQ and validating its anti-osteoarthritic activity by the in vivo model. Notable levels of phenolics and flavonoids were found in the ethanol extracts of both CQ stem (CQSE) and root (CQRE), among other solvent fractions. UPLC-MS/MS analysis of these selective extracts resulted in different classes of active compounds from both positive and negative ionization modes. By analyzing their mass spectra and fragmentation pattern, 25 active compounds were identified. The CQSE and CQRE extracts, along with the standard drug (naproxen), were further tested in mono-sodium iodoacetate-induced experimental OA animals. The modulatory effects of the test extracts were assessed by haematology, synovial and cartilage marker profiling, radiology and histopathological analysis. The in vivo findings from the biochemical and physiological studies have led to the conclusion that the CQSE extract is a good choice for the management of OA. The results were substantially better than CQ root extract and naproxen drug-treated groups. Thus, CQS has bioactive constituents, which could facilitate recovery from joint tissue damage, cellular metabolism and associated risk factors attributable to dysfunctions in OA incidence and progression.


Governing the healthy male citizen: men, masculinity and popular health in Men's Health magazine.
Recent commentators have noted the potential of newer neo-liberal discourses of health care to position responsibility for the management of well-being with the individual. Often promoted through the inculcation of risk avoidance and management, such discourses are played out in myriad settings, including the popular media. Magazines are one such media site in which diverse exhortations for the achievement of health, well-being and the perfectible body are made, and Bunton [1997. Popular health, advanced liberalism and good housekeeping magazine. In A. Petersen & Bunton R. (Eds.) Foucault, health and medicine (pp. 223-247). London: Routledge] has identified 'magazine medicine' as a significant manifestation of more dedifferentiated models of health care. Recent discussions have placed men's health high on research and policy agendas, with a concomitant interest in more popular realms. The UK magazine Men's Health (MH) is indicative of these trends, and represents a site at which discourses of men, health and masculinity are constructed. Typically reflecting neo-liberal models of health, here men are constructed as active and entrepreneurial citizens able to maintain their own health and well-being through the judicious management of risk in contexts appropriate to dominant discourses of hegemonic masculinity. Data which resulted from a critical discourse analysis of a 2-year sample (21 issues) of MH are considered and findings related to medicalisation, individualisation and risk discussed. It is suggested that magazine texts such as MH reflect newer individualised models of health care and neo-liberal strategies of health governance premised upon constructing a healthy male citizen, willing and able to take responsibility for their own well-being.


Multitargeted Flavonoid Inhibition of the Pathogenic Bacterium Staphylococcus aureus: A Proteomic Characterization.
Growth inhibition of the pathogen Staphylococcus aureus with currently available antibiotics is problematic in part due to bacterial biofilm protection. Although recently characterized natural products, including 3',4',5-trihydroxy-6,7-dimethoxy-flavone [1], 3',4',5,6,7-pentahydroxy-flavone [2], and 5-hydroxy-4',7-dimethoxy-flavone [3], exhibit both antibiotic and biofilm inhibitory activities, the mode of action of such hydroxylated flavonoids with respect to S. aureus inhibition is yet to be characterized. Enzymatic digestion and high-resolution MS analysis of differentially expressed proteins from S. aureus with and without exposure to antibiotic flavonoids (1-3) allowed for the characterization of global protein alterations induced by metabolite treatment. A total of 56, 92, and 110 proteins were differentially expressed with bacterial exposure to 1, 2, or 3, respectively. The connectivity of the identified proteins was characterized using a search tool for the retrieval of interacting genes/proteins (STRING) with multitargeted S. aureus inhibition of energy metabolism and biosynthesis by the assayed flavonoids. Identifying the mode of action of natural products as antibacterial agents is expected to provide insight into the potential use of flavonoids alone or in combination with known therapeutic agents to effectively control S. aureus infection.


Effects of oestrogen administration on vitamin B6 and tryptophan metabolism in the rat.
1. In order to assess the effects of oestrogens on the metabolism of tryptophan and vitamin B6, ovariectomized rats have been maintained on diets providing known amounts of tryptophan, nicotinamide and vitamin B6. They received oestrone sulphate, 210 micrograms/kg body-wt per d, either incorporated in the diet for 8 weeks, or by daily intraperitoneal injection for periods of 1-3 d. 2. Oestrone sulphate administration caused a slight reduction in the concentration of pyridoxal phosphate in plasma. It had no effect on the concentration of pyridoxal phosphate in liver or kidney, the urinary excretion of 4-pyridoxic acid, the activation of erythrocyte aspartate aminotransferase (L-aspartate:2-oxo-glutarate aminotransferase, EC 2. 6. 1. 1) by incubation with added pyridoxal phosphate, or the activity of pyridoxal oxidase (aldehyde:oxygen oxido-reductase, EC 18.104.22.168) in the liver. 3. Oestrone sulphate administration caused an increase in the urinary excretion of kynurenine and a reduction in the activity of liver kynureninase (L-kynurenine hydrolase, EC 184.108.40.206). It had no effect on the urinary excretion of N1-methyl nicotinamide or the concentrations of nicotinamide nucleotides in blood, liver or kidney. 4. There was a considerable excess of the apoenzyme of kynureninase in the liver. Incubation of liver homogenates with added pyridoxal phosphate led to a 4- to 5-fold increase in activity. 5. We conclude that there is no evidence of any significant effect of oestrogens on vitamin B6. It is suggested that abnormalities of tryptophan metabolism in women receiving oestrogens, which have been widely attributed to drug-induced vitamin B6 depletion, can be accounted for by inhibition of kynureninase by oestrogen metabolites.


Two-year clinical outcomes of drug-coated balloon angioplasty and angiographic predictors of restenosis among patients with de novo femoropopliteal lesions.
We analyzed the 2-year clinical outcomes of patients with de novo femoropopliteal (FP) lesions who underwent drug-coated balloon (DCB) angioplasty and the angiographic predictors of restenosis. This single-center, retrospective, and observational study evaluated 129 de novo FP lesions treated with DCB angioplasty without bailout stenting. Clinical outcomes and risk factors for loss of primary patency were analyzed using univariate and multivariate Cox proportional hazards regression models. The participants were aged 48-93 (mean: 73.6 ± 9.8) years, and 31% were women. Approximately 33% of the patients were receiving regular dialysis, and 35% of the affected limbs had critical ischemia. The mean lesion length was 132 ± 96 mm, and the mean reference vessel diameter (RVD) was 4.7 ± 0.8 mm. Forty-three (33%) limbs had chronic total occlusion of the target artery segment. Fifty-seven (44%) and 72 (56%) lesions were treated with DCB angioplasty using IN.PACT Admiral and Lutonix, respectively. The primary patency and amputation-free survival at 2 years were 59.3% and 89.5%, respectively. RVD was found to be an independent predictor of loss of primary patency. Based on the receiver operating characteristic analysis, an RVD of 4.2 mm was the best predictor of loss of primary patency at 2 years. The short-term clinical outcome of DCB angioplasty for de novo FP lesions was acceptable. Moreover, an RVD of <4.2 mm was an independent predictor of restenosis after DCB angioplasty.


Oxygen and glucose deprivation in an organotypic hippocampal slice model of the developing rat brain: the effects on N-methyl-D-aspartate subunit composition.
Organotypic hippocampal slices (OHS) are commonly used to screen for neuroprotective effects of pharmacological agents relevant to pediatric brain injury. The importance of donor rat pup age and N-methyl-D-aspartate (NMDA) receptor subunit composition have not been addressed. In this study, we evaluated the age-dependent effect of oxygen-glucose deprivation (OGD) in the developing rat brain and determined whether OGD modulates the NMDA receptor subunit composition. OHS were prepared from rat pups on postnatal days (PND) 4, 7, 14, and 21 and cultured 7 days in vitro. The slices were exposed to OGD for durations of 5-60 min. After 24 and 72 h, OHS survival and NMDA subunit composition were assessed. Cell death was evident in OHS prepared from PND 14 and 21 rat pups (P < 0.001) with OGD durations of 5 and 10 min, respectively. In OHS prepared from PND7 rat pups, neurodegeneration was not evident until 20 min OGD (P < 0.001). Exposure to OGD in OHS prepared from PND4 and PND7 rat pups was associated with a transition in the NMDA receptor subunit composition from NR2B predominant to NR2A predominant subunit composition. This in vitro neonatal rat pup investigation using OHS supports both an age and an NMDA receptor subunit composition-dependent relationship between OGD and neuronal cell death.


Scope for rotavirus vaccination in India: revisiting the scientific evidence.
Rotavirus vaccines have been developed to prevent deaths resulting from severe diarrhea of rotavirus origin. The use of vaccines as an intervention at scale to prevent and control the burden of rotavirus diarrhea is supported by the argument that prevailing public health measures such as hygiene and sanitation, breast feeding and use of ORS have failed to prevent severe dehydration resulting from diarrhea. The article reviews the existing evidence on the rationale of using rotavirus vaccine as against the feasibility of scaling it up in developing countries like India. The vaccines currently available may not cover the strains circulating in Indian population. The diversity of Rotavirus infection in the country is tremendous and since the safety, immunogenicity and efficacy data has not been collected for India, there is first a need to conduct studies to measure the extent of protection and cross-protection provided by the available vaccines for local strains, before venturing into Rotavirus vaccination program. The potential benefits of immunization have to be first vetted against the risks involved by the policymakers and other stakeholders.


New isoflavone and triterpene glycosides from soybeans.
Chemical investigation of a soybean phytochemical concentrate resulted in the isolation and identification of two new isoflavanones, dihydrodaidzin (1) and dihydrogenistin (2), a new isoflavone, 2' ',6' '-O-diacetyloninin (3), and two new triterpenoid saponins (13 and 14). Nine known isoflavonoids (4-12) and three known saponins (15-17) were also identified. Structures of the new compounds were established by a combination of extensive NMR (DEPT, DQF-COSY, HMBC, HMQC, and ROESY) studies and chemical degradation. Cytotoxic activities (ED(50)) of various extracts and selected isoflavonoids and saponins were measured against human stomach carcinoma (Hs 740.T, Hs 756 T), breast adenocarcinoma (Hs 578 T, Hs 742.T), and prostate carcinoma (DU 145, LNCaP-FGC) cell lines. Isoflavonoids 3 and 5 were more active than 1 and 2 versus at least one of the three cell lines examined, indicating the importance of the 2,3-double bond in cytotoxicity. Saponins 13, 14, and 15 were slightly more active than saponins 16 and 17, indicating that sugar attachments at position-22 enhance cytotoxic activity.


The Beneficial Role of Anchomanes difformis in STZ-Induced Reproductive Dysfunction in Male Wistar Rats.
Progression of diabetes mellitus has increasingly led to several diabetic complications. Diabetes is one of the major factors implicated in male reproductive system damage. Recent approaches such as the use of medicinal plants have been explored in the management of diabetes and associated complications. Anchomanes difformis (common name: children's umbrella) has been shown to possess anti-diabetic ability in animal model. Therefore, this study seeks to investigate the potency of Achomanes difformis in ameliorating diabetes-induced reproductive dysfunction. Type 2 diabetes was induced in male Wistar rats with 10% fructose administration for 2 weeks and an intraperitoneal injection of 40mg/kgBW of streptozotocin. Aqueous extract (200mg and 400mg/kgBW) of Anchomanes difformis leaves was administered daily for 6 weeks. The rats were randomly divided into 7 groups with a minimum of eight rats in each (8 rats in normal groups and 10 in diabetic groups). The impact of diabetes and treatment was investigated by estimating sperm concentration, motility indices, viability and morphological parameters in the normal, treatment controls and diabetic rats using CASA-SCA system. Histological examination of the testes and epididymis was performed. Diabetes induction resulted in significant decrease in sperm concentration, viability and some motility parameters with 40% abnormalities in sperm morphology. The administration of Anchomanes difformis significantly increased sperm concentration and sperm viability, while it significantly improved the percentage of morphologically normal sperm in diabetic rats. Anchomanes difformis ameliorated testicular damage such as vacuolization and loss of germinal epithelium in the diabetic-treated rats when compared to the diabetic controls. The potency Anchomanes difformis displayed against diabetic-induced damage in the reproductive system might be a new and promising tool in the management of male reproductive dysfunctions and associated complications in diabetes mellitus.


Injury pattern of road traffic accident cases attending trauma centre of tertiary care hospital of North India.
Injury is a significant global health burden and can result in mortality if not attended to on time. Trauma system refers to a collection of services provided by various super-specialties. According to the WHO-World Bank Report, RTA will rise from ninth place to the third biggest cause of mortality. The study was done at Advanced Trauma Centre (ATC) at PGIMER, a teaching hospital of north India. Study included area from most of the patient come for treatment (rural/urban) and injury patterns seen in these patients, which included mode of injury, type of injury, type of road accidents and location of injury. In maximum cases, 60.2% (245) of the mode of injury was RTA. It was seen that the maximum number of patients, 44.4%, (115) had motorbike/scooter collisions with vehicles. In most patients, the type of injury seen was 35.9% (147) head, neck and back injuries, and in maximum cases, the location of the accident site was road/street 63.2%. In our country, where the trauma delivery system is poorly developed, teaching hospitals have to bear the burden of treating many patients. No concept of emergency medicine or trauma care is in use, even in urban areas. As a result, teaching hospitals' emergency departments receive many referrals for emergency conditions.


Increased Anxiety and Depression Among Belgian Sexual Minority Groups During the First COVID-19 Lockdown-Results From an Online Survey.
The COVID-19 pandemic most likely had a negative impact on mental health. Sexual minorities are at higher risk for adverse mental outcomes such as depression, anxiety and suicidal ideation. Such mental health disparities may have exacerbated during the COVID-19 pandemic, due to restricted real-life social contact. The study aim was to examine changes in depression, anxiety and suicidal ideation among Belgian sexual minority adults between the periods before and during the first COVID-19 lockdown. We conducted an online survey, which was disseminated by community organizations throughout Belgium in April 2020. The questionnaire included two-item Generalized-Anxiety-Disorder (GAD-2) and Patient-Health-Questionnaire (PHQ-2) measures. To assess how such symptoms and other factors (e.g., loneliness) had changed, we asked to what extent these occurred before and since the lockdown. We included 965 fully completed questionnaires in the analysis. The proportions of participants screening positive for depression and anxiety were significantly higher during the lockdown than before the lockdown, based on their reported symptoms for these periods: 29.3%% vs. 13.5% (p < 0.001), and 37.1% vs. 25.7% (p < 0.001) respectively. Lonely and young participants were more likely to acquire depression. About one in five participants reported suicidal ideation. Our findings suggest that the COVID-19 pandemic has exacerbated already existing mental health disparities between sexual minority adults and the general population. These exacerbations may be the result of increased loneliness and social isolation. The results highlight the need for stimulating and strengthening social connectedness within the LGBTQI community during and in the aftermath of the COVID-19 pandemic, and the need for maintaining mental health services for such groups during pandemic restrictions.


Znf202 affects high density lipoprotein cholesterol levels and promotes hepatosteatosis in hyperlipidemic mice.
The zinc finger protein Znf202 is a transcriptional suppressor of lipid related genes and has been linked to hypoalphalipoproteinemia. A functional role of Znf202 in lipid metabolism in vivo still remains to be established. We generated mouse Znf202 expression vectors, the functionality of which was established in several in vitro systems. Next, effects of adenoviral znf202 overexpression in vivo were determined in normo- as well as hyperlipidemic mouse models. Znf202 overexpression in mouse hepatoma cells mhAT3F2 resulted in downregulation of members of the Apoe/c1/c2 and Apoa1/c3/a4 gene cluster. The repressive activity of Znf202 was firmly confirmed in an apoE reporter assay and Znf202 responsive elements within the ApoE promoter were identified. Adenoviral Znf202 transfer to Ldlr-/- mice resulted in downregulation of apoe, apoc1, apoa1, and apoc3 within 24 h after gene transfer. Interestingly, key genes in bile flux (abcg5/8 and bsep) and in bile acid synthesis (cyp7a1) were also downregulated. At 5 days post-infection, the expression of the aforementioned genes was normalized, but mice had developed severe hepatosteatosis accompanied by hypercholesterolemia and hypoalphalipoproteinemia. A much milder phenotype was observed in wildtype mice after 5 days of hepatic Znf202 overexpression. Interestingly and similar to Ldl-/- mice, HDL-cholesterol levels in wildtype mice were lowered after hepatic Znf202 overexpression. Znf202 overexpression in vivo reveals an important role of this transcriptional regulator in liver lipid homeostasis, while firmly establishing the proposed key role in the control of HDL levels.


Hepatocyte targeting of nucleic acid complexes and liposomes by a T7 phage p17 peptide.
A critical step for liver-directed gene therapy is the selective targeting of nucleic acids to hepatocytes. We have previously discovered that the proximal half of the T7 phage tail fiber protein (p17) targeted intact T7 phage and recombinant proteins to hepatocytes in vivo. In the present study, we have localized the targeting activities to a 33 amino acid sequence within the p17 coiled-coil rod domain. Given that the tail fiber domain from which the peptide was derived may form alpha and triple helical structures, biophysical studies (CD spectra and analytical ultracentrifugation) were conducted to determine the secondary and tertiary structures of the peptide. This peptide is able to target proteins, polymers, and siRNA and also particles such as DNA polyplexes and liposomes to hepatocytes. A variety of coupling strategies and chemistries were employed, thus demonstrating that this peptide is a versatile system for delivering cargo. The ability of this hepatocyte-targeting peptide to target DNA-containing particles suggests that it should be useful in the development of both nonviral and viral vectors. However, biological function of delivered cargo has not been demonstrated. This was primarily due to failure of delivered cargo to escape the endosomes. Further studies are in progress to provide functional activity of delivered nucleic acids by enabling their endosomal escape.


Coupling metabolism and chemotaxis-dependent behaviours by energy taxis receptors.
Bacteria have evolved the ability to monitor changes in various physico-chemical parameters and to adapt their physiology and metabolism by implementing appropriate cellular responses to these changes. Energy taxis is a metabolism-dependent form of taxis and is the directed movement of motile bacteria in gradients of physico-chemical parameters that affect metabolism. Energy taxis has been described in diverse bacterial species and several dedicated energy sensors have been identified. The molecular mechanism of energy taxis has not been studied in as much detail as chemotaxis, but experimental evidence indicates that this behaviour differs from metabolism-independent taxis only by the presence of dedicated energy taxis receptors. Energy taxis receptors perceive changes in energy-related parameters, including signals related to the redox and/or intracellular energy status of the cell. The best-characterized energy taxis receptors are those that sense the redox state of the electron transport chain via non-covalently bound FAD cofactors. Other receptors shown to mediate energy taxis lack any recognizable redox cofactor or conserved energy-sensing motif, and some have been suggested to monitor changes in the proton motive force. The exact energy-sensing mechanism(s) involved are yet to be elucidated for most of these energy sensors. By monitoring changes in energy-related parameters, energy taxis receptors allow cells to couple motility behaviour with metabolism under diverse environmental conditions. Energy taxis receptors thus provide fruitful models to decipher how cells integrate sensory behaviours with metabolic activities.


Increasing the Number of Underrepresented Minority Behavioral Health Researchers Partnering With Underresourced Communities: Lessons Learned From a Pilot Research Project Program.
To address critical health equity issues facing racially and ethnically diverse populations, it is essential to have researchers from similarly diverse backgrounds. Such researchers provide different perspectives that may lead to distinct research questions, novel interpretation of findings, and innovative recommendations for health promotion practice. There is a continuing need to increase the number of researchers leading health research studies who are from underrepresented minority populations (URMs). The literature demonstrates the effectiveness of mentoring for career development and the need to hone existing mentoring models. The TREE Center developed an innovative model for building capacity among early stage investigators, with a focus on URMs, to increase the inclusivity of the research pipeline. Our model involves community-engaged behavioral health research mentoring, career development, training for grantspersonship, and guidance for manuscript development and submission. A pilot project program provided opportunities for 10 early stage investigators to develop relationships with public health practitioners and other community partners, to obtain funding, to manage a complex pilot research project, and to generate preliminary data. Awardees worked with an academic mentor, a community mentor, and TREE Center faculty to conduct and disseminate their research. Lessons learned include the need to account for funding cycle timing, address challenges of recruiting URMs, consider overutilization of senior URM mentors, and overcome institutional bureaucracies that hinder transdisciplinary research across campuses. We discuss strategies for addressing these challenges. Our model is replicable and could be implemented, especially by academic programs interested in cultivating early stage URM investigators to conduct behavioral health research.


Female responsiveness to erotic films and the "ideal" erotic film from a feminine perspective.
Subjects were 20 volunteers randomly selected from all female undergraduate students attending a local university, who were 21 years of age or older, and living off campus. The stimuli consisted of seven films, of approximately 5 minutes duration each, which depicted various types of human sexual behavior. Themes included "romantic" heterosexual behavior, primarily genital heterosexual behavior, a mild, and an explicit form of group-sex behavior, a mild and explicit form of heterosexual sadomasochistic behavior, and male homosexuality. Subjects rated the degree of sexual arousal experienced for each film on a 50-point Likert scale. The order of presentation of the films for each subject was randomized, and all subjects participated individually with only a female experimenter present. Following the rating period, subjects completed the Sex Knowledge and Attitude Test (9). Finally, a structured interview was conducted to probe specific likes and dislikes with respect to the content of the films viewed. Results indicated that rank order ratings of the films, from most to least sexually stimulating, was: heterosexual, romantic; group-sex mild (two males and one female); heterosexual-genital; group-sex explicit (three males and three females); sadomasochism (mild); sadomasochism-explicit (forcing and brutal); and male homosexuality. One conclusion that the data led to was that females preferred and were significantly more sexually stimulated by films in which a male related to a female (even if the male was treating the female cruelly) than they were by homosexual stimuli involving two males together. Correlational data between the rating responses and the Sex Knowledge and Attitude Test yielded much information including significant negative correlations between ratings of both group-sex films and rejection of sexual myths. Also, significant positive correlations were found between rated degree of sexual stimulation for the heterosexual and the group-sex films and the consideration of pre- and extramarital relations as acceptable or even desirable.


Pathogenesis of the hyperlipidemia of Gram-negative bacterial sepsis may involve pathomorphological changes in liver sinusoidal endothelial cells.
The Gram-negative bacterium Pseudomonas aeruginosa is one of the most common opportunistic pathogens, especially after liver transplantation. Pathophysiological alterations of liver sinusoidal endothelial cells (LSECs) have far-reaching repercussions on the liver and on metabolism. LSECs are perforated with fenestrations, pores that facilitate the transfer of lipoproteins and macromolecules between blood and hepatocytes. Gram-negative bacterial endotoxin (lipopolysaccharide, LPS) and the P. aeruginosa toxin, pyocyanin, have marked effects on LSECs. Initial loss of LSEC porosity (defenestration) induced by P. aeruginosa pyocyanin and LPS may confer subsequent immune tolerance to circulating bacterial antigens and toxins. This review collates the known immune responses of the liver to Gram-negative bacterial toxins, with a focus on LSECs. Hyperlipidemia is an important response to Gram-negative bacterial sepsis. The mechanisms proposed for sepsis-associated hyperlipidemia include tissue lipoprotein lipase inhibition and upregulated hepatic triglyceride production. In this review, we propose defenestration of the LSECs by bacterial toxins as an additional mechanism for the hyperlipidemia of sepsis. Given the role of LSECs in hyperlipidemia and liver allograft rejection, LSEC changes induced by P. aeruginosa toxins including LPS and pyocyanin may have significant clinical implications.


Consequences of single-site mutations in the intestinal fatty acid binding protein.
The intestinal fatty acid binding protein (IFABP) is a small (15 kDa) protein consisting mostly of 10 antiparallel beta-strands (A-J) and a small helical region that serves as a portal for the ligand. Two beta-sheet structures (strands A-E and F-J) surround a cavity into which the ligand binds. In this work, we investigated how changes in the side chains of specific residues are propagated through the structure. To determine what these changes were and how they relate to changes in stability, (15)N chemical shift perturbations were measured and compared to those of the wild-type protein. Seven mutations, five of which change either valine or leucine to glycine, have been examined. All these mutants were less stable than wild-type IFABP, suggesting some structural changes. For five of the mutants, the data suggest that destabilization of a small region of the protein propagates throughout the structure, resulting in an overall decrease in stability. In two (Leu38Gly and Leu89Gly), the loss of cooperativity in the equilibrium denaturation curves suggests that the destabilization of one region may not be transmitted to other regions in a cooperative manner. It is shown that the effect of mutating hydrophobic residues is much greater than that observed upon mutation of a solvent-exposed polar residue.


Serotonin neurons modulate learning rate through uncertainty.
Regulating how fast to learn is critical for flexible behavior. Learning about the consequences of actions should be slow in stable environments, but accelerate when that environment changes. Recognizing stability and detecting change are difficult in environments with noisy relationships between actions and outcomes. Under these conditions, theories propose that uncertainty can be used to modulate learning rates ("meta-learning"). We show that mice behaving in a dynamic foraging task exhibit choice behavior that varied as a function of two forms of uncertainty estimated from a meta-learning model. The activity of dorsal raphe serotonin neurons tracked both types of uncertainty in the foraging task as well as in a dynamic Pavlovian task. Reversible inhibition of serotonin neurons in the foraging task reproduced changes in learning predicted by a simulated lesion of meta-learning in the model. We thus provide a quantitative link between serotonin neuron activity, learning, and decision making.


Prenatal diagnosis of methylmalonic aciduria by analysis of organic acids and total homocysteine in amniotic fluid.
Methylmalonic aciduria (MMA) is the most frequent disease of organic aciduria in China. Various biochemical strategies are followed for the prenatal diagnosis of MMA. However, since fetuses affected by MMA have decreased excretion of methylmalonic acid, the difficulties of prenatal biochemical diagnosis are obvious. Gas chromatography mass spectrometry (GC/MS) and tandem mass spectrometry (ESI/MS/MS) have allowed us to identify the disease in affected fetuses. The aim of this study was to determine the value of analysis of organic acids and total homocysteine in amniotic fluid in prenatal diagnosis of MMA. The clinical diagnoses and outcomes of nine probands with MMA and the prenatal diagnoses based on biochemical analysis of nine fetuses at risk for MMA were investigated. Amniotic fluid samples from pregnancies at risk for MMA and metabolically normal pregnancies were obtained at 16 - 24 weeks of gestation. Methylmalonic acid and methylcitric acid were measured by GC/MS, propionylcarnitine was analyzed by ESI/MS/MS, and total homocysteine was determined by fluorescence polarization immunoassay. In two pregnancies, high levels of methylmalonic acid, methylcitric acid, propionylcarnitine, and total homocysteine indicated combined MMA and homocysteinemia in the fetuses. One of the mothers continued pregnancy and received cobalamin supplement as prenatal treatment, and the other terminated her pregnancy. In one pregnancy, significantly elevated levels of methylmalonic acid, methylcitric acid, and propionylcarnitine, and normal level of total homocysteine was found indicating isolated MMA in the fetus; abortion was performed on this case. In the other six pregnancies, all the levels of the above mentioned metabolites were normal suggesting that the fetuses were not affected by MMA. The diagnoses were confirmed after delivery by testing urinary organic acids and plasma total homocysteine. The metabolic abnormalities of MMA occur early in gestation. The level of total homocysteine in amniotic fluid may be an additional indicator of fetal combined MMA and homocysteinemia. Determination of total homocysteine level in amniotic fluid may become a convenient and reliable method for prenatal diagnosis of the disease.


Interrelationships in trace-element metabolism in metal toxicities in a cobalt-resistant strain of Neurospora crassa.
A strain of Neurospora crassa was isolated by training the mould to grow on media containing high concentrations of Co(2+). This strain, the Co(R) strain, exhibited approximately tenfold the resistance of the parent strain to Co(2+) and Ni(2+) but not to Zn(2+) or Cu(2+). Co(2+) toxicity in the Co(R) strain was reversed by Mg(2+) but not by Fe(3+). Also, Co(2+) did not affect iron metabolism in this strain. It is suggested that the mechanism of resistance in the Co(R) strain involves an alteration in the pattern of iron metabolism such that the latter is no longer adversely affected by toxic concentrations of Co(2+). The Co(R) strain is genetically stable and is most probably a result of a resistance mutation in N. crassa induced by Co(2+).


The v-abl, c-fms, or v-myc oncogene induces gamma radiation resistance of hematopoietic progenitor cell line 32d cl 3 at clinical low dose rate.
A variety of viral and cellular oncogenes have been described with differing mechanisms of action but with the common property of inducing morphologic alteration of cells in culture. Subclonal lines of oncogene expressing cells have been shown to produce tumors in vivo. Expression of the N-ras oncogene in embryo fibroblast NIH/3T3 cells has been demonstrated to increase radioresistance in vitro, and these results have been confirmed and extended to human cell lines expressing the c-raf oncogene. In the present report, we have examined the effects of expression of the c-fms, v-abl, or v-myc oncogene in a clonal hematopoietic progenitor cell line 32D cl 3. The 32D cell line is nonmalignant in vivo and is dependent upon a source of Interleukin-3 (IL-3) for growth in vitro. The radiation survival of 32D cl 3 cells transfected and expressed in the c-fms oncogene showed significant increase in the radioresistance at both 5 cGy/min and 116 cGy/min. A clone of 32D cl 3 transfected and expressing the v-myc oncogene demonstrated increased radioresistance at both dose rates. Results of split dose experiments suggested significant repair of sublethal irradiation damage of 32D-v-abl cells. Results were compared with expression of the same v-abl oncogene in the NIH/3T3 embryo fibroblast cell line. The data demonstrate that gamma irradiation resistance is significantly increased by each oncogene expressed in 32D cl 3 cells. The data on cell line 32D cl 3 may correlate with the radioresistance of v-abl expressing human hematopoietic cell malignancies treated by irradiation therapy.


Thalamic deep brain stimulation modulates cycles of seizure risk in epilepsy.
Chronic brain recordings suggest that seizure risk is not uniform, but rather varies systematically relative to daily (circadian) and multiday (multidien) cycles. Here, one human and seven dogs with naturally occurring epilepsy had continuous intracranial EEG (median 298 days) using novel implantable sensing and stimulation devices. Two pet dogs and the human subject received concurrent thalamic deep brain stimulation (DBS) over multiple months. All subjects had circadian and multiday cycles in the rate of interictal epileptiform spikes (IES). There was seizure phase locking to circadian and multiday IES cycles in five and seven out of eight subjects, respectively. Thalamic DBS modified circadian (all 3 subjects) and multiday (analysis limited to the human participant) IES cycles. DBS modified seizure clustering and circadian phase locking in the human subject. Multiscale cycles in brain excitability and seizure risk are features of human and canine epilepsy and are modifiable by thalamic DBS.


The actin-binding domain of Slac2-a/melanophilin is required for melanosome distribution in melanocytes.
Melanosomes containing melanin pigments are transported from the cell body of melanocytes to the tips of their dendrites by a combination of microtubule- and actin-dependent machinery. Three proteins, Rab27A, myosin Va, and Slac2-a/melanophilin (a linker protein between Rab27A and myosin Va), are known to be essential for proper actin-based melanosome transport in melanocytes. Although Slac2-a directly interacts with Rab27A and myosin Va via its N-terminal region (amino acids 1 to 146) and the middle region (amino acids 241 to 405), respectively, the functional importance of the putative actin-binding domain of the Slac2-a C terminus (amino acids 401 to 590) in melanosome transport has never been elucidated. In this study we showed that formation of a tripartite protein complex between Rab27A, Slac2-a, and myosin Va alone is insufficient for peripheral distribution of melanosomes in melanocytes and that the C-terminal actin-binding domain of Slac2-a is also required for proper melanosome transport. When a Slac2-a deletion mutant (DeltaABD) or point mutant (KA) that lacks actin-binding ability was expressed in melanocytes, the Slac2-a mutants induced melanosome accumulation in the perinuclear region, possibly by a dominant negative effect, the same as the Rab27A-binding-defective mutant of Slac2-a or the myosin Va-binding-defective mutant. Our findings indicate that Slac2-a organizes actin-based melanosome transport in cooperation with Rab27A, myosin Va, and actin.


Nine pheochromocytomas in the same patient. Final mapping with ultrasound and angiography.
A 37-year-old man presented with hypertension and elevated urine catecholamine. Ultrasound scanning revealed a solid tumour of the right adrenal gland and two solid tumours in the retroperitoneum. The findings were confirmed with computed tomography and abdominal angiography. At surgery only the tumour of the right adrenal gland was removed. The histopathologic diagnosis was pheochromocytoma. Postoperatively the symptoms and biochemistry were unchanged and the patient was referred for further treatment. At ultrasonography and abdominal aortography 6 remaining tumours were demonstrated. Surgery was performed and 8 pheochromocytomas were extirpated (3 were closely spaced small tumours in a conglomerate corresponding to one of the visualized tumour sites). On histopathologic examination no signs of invasive growth were found. The patient recovered completely. The blood pressure was still normal 2 1/2 years later. Angiography and non-invasive examination of the entire abdomen and pelvis should be routine when pheochromocytomas are searched for.


The action of thrombin on modified fibrinogen.
The modification of canine fibrinogen with citraconic anhydride modified the epsilon-amino groups of the fibrinogen and at the same time generated additional negative charges into the protein. The addition of thrombin to the modified fibrinogen did not induce polymerization; however, the fibrinopeptide was released at a faster rate than from the unmodified fibrinogen. The physical properties of the citraconylated fibrinogen were markedly altered by the modification of 50-60 lysine residues in one hour. A modified fibrinopeptide-A was released by thrombin from the modified fibrinogen and was electrophoretically more anionic than the unmodified fibrinopeptide-A. Edman analysis confirmed the modification of the lysine residue present in the peptide. The rate of removal of citraconylated fibrinopeptide-A from modified fibrinogen by thrombin was 30 to 40 percent greater than the cleavage of unmodified fibrinopeptide-A from unmodified fibrinogen. However, the modification of 60 or more lysine residues in the fibrinogen produced a decrease in the rate of cleavage of citraconylated fibrinopeptide-A. The results suggest that additional negative charge in the vicinity of the attachment of fibrinopeptide-A to canine fibrinogen aids in the removal of the peptide by thrombin.


[Heparin-induced thrombocytopenia: Update].
Heparin-induced thrombocytopenia (HIT) is a serious complication of heparin therapy. It is due to the synthesis of antibodies most often directed against platelet factor 4 (FP4) modified by heparin (H). HIT is manifested by a platelet count fall, associated with a high risk of venous or arterial thrombosis. The diagnosis of HIT is based on the assessment of clinical probability (4Ts score or change in platelet count after cardiac surgery) and the demonstration of heparin-modified anti-FP4 antibodies (FP4/H). If the immunological tests are positive, functional tests should be performed. In case of suspicion of HIT, it is necessary to urgently stop heparin therapy, to perform a doppler ultrasound of the lower limbs, and to prescribe an alternative anticoagulation agent at a curative dose. Currently, danaparoid sodium and argatroban are authorized. The diagnosis and management of HIT remain complex and requires multidisciplinary collaboration.


Whole exome sequencing reveals concomitant mutations of multiple FA genes in individual Fanconi anemia patients.
Fanconi anemia (FA) is a rare inherited genetic syndrome with highly variable clinical manifestations. Fifteen genetic subtypes of FA have been identified. Traditional complementation tests for grouping studies have been used generally in FA patients and in stepwise methods to identify the FA type, which can result in incomplete genetic information from FA patients. We diagnosed five pediatric patients with FA based on clinical manifestations, and we performed exome sequencing of peripheral blood specimens from these patients and their family members. The related sequencing data were then analyzed by bioinformatics, and the FANC gene mutations identified by exome sequencing were confirmed by PCR re-sequencing. Homozygous and compound heterozygous mutations of FANC genes were identified in all of the patients. The FA subtypes of the patients included FANCA, FANCM and FANCD2. Interestingly, four FA patients harbored multiple mutations in at least two FA genes, and some of these mutations have not been previously reported. These patients' clinical manifestations were vastly different from each other, as were their treatment responses to androstanazol and prednisone. This finding suggests that heterozygous mutation(s) in FA genes could also have diverse biological and/or pathophysiological effects on FA patients or FA gene carriers. Interestingly, we were not able to identify de novo mutations in the genes implicated in DNA repair pathways when the sequencing data of patients were compared with those of their parents. Our results indicate that Chinese FA patients and carriers might have higher and more complex mutation rates in FANC genes than have been conventionally recognized. Testing of the fifteen FANC genes in FA patients and their family members should be a regular clinical practice to determine the optimal care for the individual patient, to counsel the family and to obtain a better understanding of FA pathophysiology.


Effects of anesthetics and electrical stimulation on nigrostriatal dopaminergic neurons.
Induction of anesthesia with pentobarbital (40 mg/kg), urethane (1.8 g/kg), chloral hydrate (400 mg/kg) or ketamine (150 mg/kg) followed by a 20-min sham implantation of a bipolar electrode in substantia nigra fibers caused a marked, ipsilateral increase in caudate dihydroxyphenylacetic acid (DO-PAC). Conversely, no sham stimulation was found after gamma-butyrolactone (500 mg/kg) anesthesia or in a paralyzed, artificially respired preparation. By using gamma-butyrolactone anesthesia, the nigrostriatal fibers were electrically stimulated (25 Hz, 1.5 msec duration) at differing current intensities (40, 80, 100, 120 and 180 microA). The maximum increase in caudate DOPAC occurred at 120 microA. Electrical stimulation (100 microA, 25 Hz, 1.5 msec duration) in the paralyzed preparation yielded similar increases in caudate DOPAC. By using a 50% effective current (100 microA), variation of the frequency (5, 10, 25 and 125 Hz) produced a maximum in caudate DOPAC at 10 Hz. These data reveal an interaction between many anesthetics and physical disruption of dopaminergic neurons, as well as provide an in vivo model of stimulus-dependent dopamine release.


Effect of milk thistle, Silybium marianum, extract on toxicity, development, nutrition, and enzyme activities of the small white butterfly, Pieris rapae.
The methanolic extract of milk thistle, Silybium marianum L. (Asterales: Asteraceae), was investigated for its effects on the mortality, growth, feeding indices, enzymatic activity, and levels of non-enzymatic molecules of the small white butterfly, Pieris rapae L. (Lepidoptera: Pieridae), a pest of cruciferous plants. Feeding indices including approximate digestibility (AD), efficiency of conversion of digested food (ECD), efficiency of conversion of ingested food (ECI), relative growth rate (RGR), and relative consumption rate (RCR) were measured. These indices were variously affected: the RGR, RCR, and AD decreased, but the ECD and ECI increased. The LC50 and LC25 values were estimated as 2.94% and 1.20%, respectively. At the lowest concentration of S. marianum extract (0.625%), the feeding deterrence index was 40.48%. The duration of the pupal stage and the rate of larval growth decreased. These changes may be due to alterations in metabolic activity, such as the increase in alkaline phosphatase activity, which is likely involved in detoxification. Additionally, the activities of alanine aminotransferase and aspartate aminotransferase, which are key components of amino acid catabolism, decreased. The amount of glucose (an energy source) and uric acid (the excreted end product) increased, while total protein (another energy source) and cholesterol decreased. These results indicate that this plant possesses potential secondary metabolites that may be useful for the future study of the control of insect pests.


Identifying the Potential Differentially Expressed miRNAs and mRNAs in Osteonecrosis of the Femoral Head Based on Integrated Analysis.
Osteonecrosis of the femoral head is a common disease of the hip that leads to severe pain or joint disability. We aimed to identify potential differentially expressed miRNAs and mRNAs in osteonecrosis of the femoral head. The data of miRNA and mRNA were firstly downloaded from the database. Secondly, the regulatory network of miRNAs-mRNAs was constructed, followed by function annotation of mRNAs. Thirdly, an in vitro experiment was applied to validate the expression of miRNAs and targeted mRNAs. Finally, GSE123568 dataset was used for electronic validation and diagnostic analysis of targeted mRNAs. Several regulatory interaction pairs between miRNA and mRNAs were identified, such as hsa-miR-378c-WNT3A/DACT1/CSF1, hsa-let-7a-5p-RCAN2/IL9R, hsa-miR-28-5p-RELA, hsa-miR-3200-5p-RELN, and hsa-miR-532-5p-CLDN18/CLDN10. Interestingly, CLDN10, CLDN18, CSF1, DACT1, IL9R, RCAN2, RELN, and WNT3A had the diagnostic value for osteonecrosis of the femoral head. Wnt signaling pathway (involved WNT3A), chemokine signaling pathway (involved RELA), focal adhesion and ECM-receptor interaction (involved RELN), cell adhesion molecules (CAMs) (involved CLDN18 and CLDN10), cytokine-cytokine receptor interaction, and hematopoietic cell lineage (involved CSF1 and IL9R) were identified. The identified differentially expressed miRNAs and mRNAs may be involved in the pathology of osteonecrosis of the femoral head.


The expression of virulence during double infections by different parasites with conflicting host exploitation and transmission strategies.
In many natural populations, hosts are found to be infected by more than one parasite species. When these parasites have different host exploitation strategies and transmission modes, a conflict among them may arise. Such a conflict may reduce the success of both parasites, but could work to the benefit of the host. For example, the less-virulent parasite may protect the host against the more-virulent competitor. We examine this conflict using the waterflea Daphnia magna and two of its sympatric parasites: the blood-infecting bacterium Pasteuria ramosa that transmits horizontally and the intracellular microsporidium Octosporea bayeri that can concurrently transmit horizontally and vertically after infecting ovaries and fat tissues of the host. We quantified host and parasite fitness after exposing Daphnia to one or both parasites, both simultaneously and sequentially. Under conditions of strict horizontal transmission, Pasteuria competitively excluded Octosporea in both simultaneous and sequential double infections, regardless of the order of exposure. Host lifespan, host reproduction and parasite spore production in double infections resembled those of single infection by Pasteuria. When hosts became first vertically (transovarilly) infected with O. bayeri, Octosporea was able to withstand competition with P. ramosa to some degree, but both parasites produced less transmission stages than they did in single infections. At the same time, the host suffered from reduced fecundity and longevity. Our study demonstrates that even when competing parasite species utilize different host tissues to proliferate, double infections lead to the expression of higher virulence and ultimately may select for higher virulence. Furthermore, we found no evidence that the less-virulent and vertically transmitting O. bayeri protects its host against the highly virulent P. ramosa.


Long-term survival following in-hospital cardiac arrest: A matched cohort study.
Each year, 200,000 patients undergo an in-hospital cardiac arrest (IHCA), with approximately 15-20% surviving to discharge. Little is known, however, about the long-term prognosis of these patients after discharge. Previous efforts to describe out-of-hospital survival of IHCA patients have been limited by small sample sizes and narrow patient populations A single institution matched cohort study was undertaken to describe mortality following IHCA. Patients surviving to discharge following an IHCA between 2008 and 2010 were matched on age, sex, race and hospital admission criteria with non-IHCA hospital controls and follow-up between 9 and 45 months. Kaplan-Meier curves and Cox PH models assessed differences in survival. Of the 1262 IHCAs, 20% survived to hospital discharge. Of those discharged, survival at 1 year post-discharge was 59% for IHCA patients and 82% for controls (p<0.0001). Hazard ratios (IHCA vs. controls) for mortality were greatest within the 90 days following discharge (HR=2.90, p<0.0001) and decreased linearly thereafter, with those surviving to one year post-discharge having an HR for mortality below 1.0. Survival after discharge varied amongst IHCA survivors. When grouped by discharge destination, out of hospital survival varied; in fact, IHCA patients discharged home without services demonstrated no survival difference compared to their non-IHCA controls (HR 1.10, p=0.72). IHCA patients discharged to long-term hospital care or hospice, however, had a significantly higher mortality compared to matched controls (HR 3.91 and 20.3, respectively; p<0.0001). Among IHCA patients who survive to hospital discharge, the highest risk of death is within the first 90 days after discharge. Additionally, IHCA survivors overall have increased long-term mortality vs. Survival rates were varied widely with different discharge destinations, and those discharged to home, skilled nursing facilities or to rehabilitation services had survival rates no different than controls. Thus, increased mortality was primarily driven by patients discharged to long-term care or hospice.


Voltage-gated potassium channel blocker 4-aminopyridine induces glioma cell apoptosis by reducing expression of microRNA-10b-5p.
Accumulating evidence has demonstrated that voltage-gated potassium channels (Kv channels) were associated with regulating cell proliferation and apoptosis in tumor cells. Our previous study proved that the Kv channel blocker 4-aminopyridine (4-AP) could inhibit cell proliferation and induce apoptosis in glioma. However, the precise mechanisms were not clear yet. MicroRNAs (miRNAs) are small noncoding RNAs that act as key mediators in the progression of tumor, so the aim of this study was to investigate the role of miRNAs in the apoptosis-promoting effect of 4-AP in glioma cells. Using a microRNA array, we found that 4-AP altered the miRNA expression in glioma cells, and the down-regulation of miR-10b-5p induced by 4-AP was verified by real-time PCR. Transfection of miR-10b-5p mimic significantly inhibited 4-AP-induced caspases activation and apoptosis. Moreover, we verified that apoptosis-related molecule Apaf-1 was the direct target of miR-10b-5p. Furthermore, miR-10b-5p mimic significantly inhibited 4-AP-induced up-regulation of Apaf-1 and its downstream apoptosis-related proteins, such as cleaved caspase-3. In conclusion, Kv channel blocker 4-AP may exert its anti-tumor effect by down-regulating the expression of miR-10b-5p and then raised expression of Apaf-1 and its downstream apoptosis-related proteins. Current data provide evidence that miRNAs play important roles in Kv channels-mediated cell proliferation and apoptosis.


Genomic characterization of the sheep vasopressin V1a receptor gene and promoter, with assignment to bands q23-24 of sheep chromosome 3 and cattle chromosome 5.
Arginine vasopressin interacts with the vasopressin type 1a receptor (V1aR) to initiate physiological effects such as vasoconstriction of blood vessels and glycogenolysis. AVP is also involved in central nervous effects such as body homeostasis and blood pressure control. The complete genomic organization of the sheep V1aR gene has been determined, including the presence of one major and two minor transcriptional start sites at -321, -206 and -91bp respectively, relative to the ATG codon. Another more distal minor transcriptional start site was also localized between nucleotides -997 and -892 relative to the ATG codon. One intron exists in the sheep V1aR gene and potential cis- and trans- acting sites were identified in the sheep V1aR promoter. The promoter was also compared to the rat V1aR promoter. The sheep V1aR promoter displays features typical of housekeeping genes, although tissue-specific expression does not support this. V1aR mRNA is absent in the adult sheep liver but not the kidney. One copy of the V1aR gene exists in the sheep genome, which was localized to chromosome 3q23-24, and to the homoeologous position, 5q23-24 in cattle.


[Ratings and complications with minimally invasive percutaneous tracheotomy].
Indication, timing and technique of tracheostomy in intensive care medicine have changed significantly during the last decade. Benefits and risks of percutaneous dilatational tracheostomy are reviewed in the current literature. Typical complications are misplacement of the tracheal cannula, injury to the trachea, trachealstenosis, rupture of the cuff-ballon and death. The results of a questionnaire about the use, timing and complications of percutaneous dilatational tracheostomy at 651 German intensive care units are presented. In Germany tracheostomy is performed within 20 days in 75% of patients on mechanical ventilation, following the recommendation of the Consensus Conference on Artificial Airways in Patients Receiving Mechanical Ventilation from 1989. Percutaneous dilatational tracheostomy in Germany is performed following the methods described by Ciaglia in 58% of intensive care units, Griggs in 35% and Fantoni in 7%.


Infrared (810-nm) low-level laser therapy on rat experimental knee inflammation.
Arthritis of the knee is the most common type of joint inflammatory disorder and it is associated with pain and inflammation of the joint capsule. Few studies address the effects of the 810-nm laser in such conditions. Here we investigated the effects of low-level laser therapy (LLLT; infrared, 810-nm) in experimentally induced rat knee inflammation. Thirty male Wistar rats (230-250 g) were anesthetized and injected with carrageenan by an intra-articular route. After 6 and 12 h, all animals were killed by CO(2) inhalation and the articular cavity was washed for cellular and biochemical analysis. Articular tissue was carefully removed for real-time PCR analysis in order to evaluate COX-1 and COX-2 expression. LLLT was able to significantly inhibit the total number of leukocytes, as well as the myeloperoxidase activity with 1, 3, and 6 J (Joules) of energy. This result was corroborated by cell counting showing the reduction of polymorphonuclear cells at the inflammatory site. Vascular extravasation was significantly inhibited at the higher dose of energy of 10 J. Both COX-1 and 2 gene expression were significantly enhanced by laser irradiation while PGE(2) production was inhibited. Low-level laser therapy operating at 810 nm markedly reduced inflammatory signs of inflammation but increased COX-1 and 2 gene expression. Further studies are necessary to investigate the possible production of antiinflammatory mediators by COX enzymes induced by laser irradiation in knee inflammation.


Applications of nonlinear ultrasonics to the NDE of material degradation.
Nonlinear ultrasonics is suggested as a new approach for the effective evaluation of material degradation. As its quantification, the parameter beta is introduced on the basis of nonlinear elasticity, and a new method to measure the parameter beta using bispectral analysis is proposed. Then, the correlation between beta and material degradation is investigated. From the results for several mild steel (SS41, SS45) specimens that were degraded by stretching and cyclic loads, it was confirmed that the parameter beta has a strong correlation with material degradation. As another practical application, the evaluation of the aging degradation in a high temperature material is tried. For this, Cr-Mo-V specimens that are generally used in turbine rotors in power plants were prepared, and the variation of beta caused by aging time was investigated. For comparison, the fracture appearance transition temperature (FATT) of the specimen was measured, and its behavior showed good agreement with beta. In addition, for all of the experiments, no noticeable change in attenuation and sound velocity in the same specimens with change of degradation were observed. From these results, it may be concluded that nonlinear ultrasonics could be applied to the quantitative evaluation of material degradation.


Coeliac disease in adults. Presentation and management.
Coeliac disease, also known as gluten-sensitive enteropathy or non-tropical sprue, is a relatively uncommon condition. The dietary presence of gliadin, an alcohol-soluble subfraction of gluten, in immunologically susceptible hosts will lead to small intestinal mucosal inflammation and subsequent mucosal villous atrophy which results in nutrient and vitamin malabsorption. The symptomatic presentations of patients with coeliac disease are related to this malabsorption process which can be reversed in the vast majority of patients with a gluten-free diet.


Myalgic Encephalomyelitis or What? The International Consensus Criteria.
Myalgic encephalomyelitis (ME) is a neuromuscular disease with two distinctive types of symptoms (muscle fatigability or prolonged muscle weakness after minor exertion and symptoms related to neurological disturbance, especially of sensory, cognitive, and autonomic functions) and variable involvement of other bodily systems. Chronic fatigue syndrome (CFS), introduced in 1988 and re-specified in 1994, is defined as (unexplained) chronic fatigue accompanied by at least four out of eight listed (ill-defined) symptoms. Although ME and CFS are two distinct clinical entities (with partial overlap), CFS overshadowed ME for decades. In 2011, a panel of experts recommended abandoning the label CFS and its definition and proposed a new definition of ME: the International Consensus Criteria for ME (ME-ICC). In addition to post-exertional neuroimmune exhaustion (PENE), a mandatory feature, a patient must experience at least three symptoms related to neurological impairments; at least three symptoms related to immune, gastro-intestinal, and genitourinary impairments; and at least one symptom related to energy production or transportation impairments to meet the diagnosis of ME-ICC. A comparison between the original definition of ME and the ME-ICC shows that there are some crucial differences between ME and ME-ICC. Muscle fatigability, or long-lasting post-exertional muscle weakness, is the hallmark feature of ME, while this symptom is facultative for the diagnosis under the ME-ICC. PENE, an abstract notion that is very different from post-exertional muscle weakness, is the hallmark feature of the ME-ICC but is not required for the diagnosis of ME. The diagnosis of ME requires only two type of symptoms (post-exertional muscle weakness and neurological dysfunction), but a patient has to experience at least eight symptoms to meet the diagnosis according to the ME-ICC. Autonomic, sensory, and cognitive dysfunction, mandatory for the diagnosis of ME, are not compulsory to meet the ME-ICC subcriteria for 'neurological impairments'. In conclusion, the diagnostic criteria for ME and of the ME-ICC define two different patient groups. Thus, the definitions of ME and ME-ICC are not interchangeable.


Simulation of nonylphenol degradation in leafy vegetables using a deuterated tracer.
In this study, the residues of nonylphenol, an endocrine-disrupting chemical, were investigated in two types of leafy vegetables, leek and cabbage, that are commonly consumed in eight provinces in China. Nonylphenol was detected in 43.50% of the 223 samples, at concentrations up to 51.90 μg kg(-1). The isotopic tracer 4-n-NP-2,3,5,6-d4-OD was chosen to simulate the degradation of nonylphenol in cabbage. More than 80% of the tracer was degraded in the first 7 days, and most of it had vanished by the end of the 35-day experimental period. A preliminary degradation equation was established to describe the variation of nonylphenol residues in leafy vegetables, and the amounts of nonylphenol in four pesticides were analyzed. The results indicated that nonylphenol residues in leafy vegetables are probably derived from the applied pesticides. The potential risk of pesticide adjuvants as well as that of the active ingredient in pesticides should be taken into consideration when recommending a safety interval period.


Increased frequency of rare missense PPP1R3B variants among Danish patients with type 2 diabetes.
PPP1R3B has been suggested as a candidate gene for monogenic forms of diabetes as well as type 2 diabetes (T2D) due to its association with glycaemic trait and its biological role in glycogen synthesis. To study if rare missense variants in PPP1R3B increase the risk of maturity onset diabetes of the young (MODY), T2D or affect measures of glucose metabolism. Targeted resequencing of PPP1R3B was performed in 8,710 samples; MODY patients with unknown etiology (n = 54), newly diagnosed patients with T2D (n = 2,930) and population-based control individuals (n = 5,726, of whom n = 4,569 had normal glucose tolerance). All population-based sampled individuals were examined using an oral glucose tolerance test. Among n = 396 carriers, we identified twenty-three PPP1R3B missense mutations, none of which segregated with MODY. The burden of likely deleterious PPP1R3B variants was significantly increased with a total of 17 carriers among patients with T2D (0.58% (95% CI: 0.36-0.93)) compared to 18 carriers among non-diabetic individuals (0.31% (95% CI: 0.20-0.49)), resulting in an increased risk of T2D (OR (95% CI) = 2.57 (1.14-5.79), p = 0.02 (age and sex adjusted)). Furthermore, carriers with diabetes had less abdominal fat and a higher serum concentration of LDL-cholesterol compared to patients with T2D without rare missense PPP1R3B variants. In addition, non-diabetic carriers had a higher birth weight compared to non-carriers. Rare missense PPP1R3B variants may predispose to T2D.


Searching for magnetism in hydrogenated graphene: using highly hydrogenated graphene prepared via Birch reduction of graphite oxides.
Fully hydrogenated graphene (graphane) and partially hydrogenated graphene materials are expected to possess various fundamentally different properties from graphene. We have prepared highly hydrogenated graphene containing 5% wt of hydrogen via Birch reduction of graphite oxide using elemental sodium in liquid NH3 as electron donor and methanol as proton donor in the reduction. We also investigate the influence of preparation method of graphite oxide, such as the Staudenmaier, Hofmann or Hummers methods on the hydrogenation rate. A control experiment involving NaNH2 instead of elemental Na was also performed. The materials were characterized in detail by electron microscopy, infrared spectroscopy, X-ray photoelectron spectroscopy, Raman spectroscopy both at room and low temperatures, X-ray fluorescence spectroscopy, inductively coupled plasma optical emission spectroscopy, combustible elemental analysis and electrical resistivity measurements. Magnetic measurements are provided of bulk quantities of highly hydrogenated graphene. In the whole temperature range up to room temperature, the hydrogenated graphene exhibits a weak ferromagnetism in addition to a contribution proportional to field that is caused not only by diamagnetism but also likely by an antiferromagnetic influence. The origin of the magnetism is also determined to arise from the hydrogenated graphene itself, and not as a result of any metallic impurities.


Lentivirus-mediated RNA interference targeting the long noncoding RNA HOTAIR inhibits proliferation and invasion of endometrial carcinoma cells in vitro and in vivo.
The overexpression of long noncoding RNA HOTAIR is associated with various aggressive solid carcinomas. However, its relationship with endometrial carcinoma has not been reported. The present study aimed to investigate the expression of the long noncoding RNA HOTAIR in endometrial carcinoma, its relationship with the carcinoma's clinicopathologic features, and the biological function of HOTAIR in regulating endometrial cancer cell proliferation and invasion in vitro and in vivo. The expression of HOTAIR was detected in different tissues and cell lines by real-time PCR. Lentivirus-mediated HOTAIR-specific shRNAvectors were transfected into endometrial cancer HEC-1A cells. Cell proliferation and colony formation were examined by CCK-8 assays and colony formation assays, respectively. Invasion and migration were examined by Transwell assays. Flow cytometry assay was used to examine the cell cycle. In addition, xenograft model assays were performed to analyze the growth of endometrial cancer cells in vivo. Our data showed that HOTAIR expression was higher in endometrial cancer cells and tissues than in normal endometrial tissues. HOTAIR expression was closely related to the tumor stage (P = 0.045), myometrial invasion (P = 0.014), and lymph node metastasis (P = 0.033). The down-regulation of HOTAIR resulted in a significant inhibition of cell proliferation, migration, and invasion and in cell cycle arrest at the G0/G1 phase. Furthermore, HOTAIR depletion significantly suppressed the endometrial cancer tumorigenesis in vivo. This study is the first to suggest that HOTAIR plays an important role in the carcinogenesis of endometrial cancer. Targeting HOTAIR may be a novel therapeutic strategy for endometrial cancer.


Adipose tissue macrophages, low grade inflammation and insulin resistance in human obesity.
Obesity was first described as a low-grade inflammatory condition more than a decade ago. However, it is only relatively recently that obese individuals have been described with increased macrophage infiltration of adipose tissue, as well as an increase in the number of "M1" or "classically activated" macrophages. Furthermore, macrophages have been identified as the primary source of many of the circulating inflammatory molecules that are detected in the obese state and are postulated to be causal both in the development of insulin resistance and in the progression to type 2 diabetes. There is also novel evidence to suggest that macrophages inhibit adipocyte differentiation, potentially leading to adipocyte hypertrophy, altered secretion of adipokines and ectopic storage of lipid within liver, muscle and other non-adipose tissues. Currently, it is not clear what causes increased macrophage infiltration of adipose tissue in obese individuals. Theories include altered signalling by adipocytes, nutritional induction of metabolic endotoxemia or reduced angiogenesis and local adipose cell hypoxia. Importantly, PPAR-gamma agonists have been shown to alter macrophage phenotype to "M2" or an "alternatively activated" anti-inflammatory phenotype and may induce macrophage specific cell death. Consequently, excitement surrounds the potential for specific inhibition of macrophage infiltration of adipose tissue via pharmacotherapy for obese patients and more particularly as adjunct therapy to improve insulin sensitivity in obese individuals with insulin resistance and overt type 2 diabetes.


Childbirth Self-Efficacy Inventory: psychometric testing of the Spanish version.
This paper is a report of a study of the psychometric properties of the Spanish version of the Childbirth Self-efficacy Inventory. Enhancing the confidence of women in their ability to cope with pain in labour is a key element of effective childbirth education programs. The Childbirth Self-efficacy Inventory is a suitable instrument for testing hypotheses about the development of self-efficacy for childbirth. Analysing its psychometric properties in the light of Bandura's Self-Efficacy Theory is relevant for the validation of the instrument. Translation and back-translation of the questionnaire took place in 2002 using a pilot sample of 12 women. The study was conducted using a sample of 146 childbearing women recruited from different national healthcare centres in Spain during the last 6 months of 2003. The psychometric study was carried out using the Statistical Package for Social Sciences. Principal components analysis supported the ability of the Self-Efficacy Inventory to distinguish between outcome and self-efficacy expectancies. Internal Consistency Reliability of the Inventory was found adequate. Previous knowledge was identified as the most important factor influencing the confidence of a woman to coping with labour. Results are consistent with previous findings, and contribute to give support to the reliability and validity of the Inventory. The findings provide an additional and useful insight into practical ways of enhancing a woman's confidence to coping with childbirth: self-efficacy can be strengthened through persuasion by healthcare professionals, childbirth educators, and family and friends who offer support to the pregnant woman.


Purine nucleotides stimulate DNA synthesis in kidney epithelial cells in culture.
Adenine nucleotides infused into animals with acute renal failure appear to enhance recovery of kidney function and structure. To determine whether these compounds could act by a direct effect on renal cell metabolism, their capacity to stimulate DNA synthesis was evaluated in cultures of monkey kidney epithelial cells (BSC-1 line). AMP and ADP enhanced DNA synthesis by threefold more than was previously observed with other mitogens for these cells. Guanosine and inosine and their nucleotides and adenosine and ATP were also mitogenic but to a lesser extent, whereas pyrimidine derivatives were ineffective. In the presence of AMP, autoradiography of [3H]thymidine-labeled cells indicated that a greater number of cells entered the S phase of the cell cycle, and assessment of cell number revealed increased multiplication. The mitogenic effect of adenine nucleotides was not reproduced by agents that raise the cellular content of cAMP and was serum independent. Adenine nucleotides did not alter DNA synthesis when added to cultures of mouse fibroblasts. These results indicate that provision of exogenous purine nucleosides and nucleotides stimulate DNA synthesis in renal epithelial cells in culture.


Insulin, glucose and hepatocellular carcinoma risk in male hepatitis B carriers: results from 17-year follow-up of a population-based cohort.
This study aimed to investigate the association of fasting insulin and glucose levels with hepatocellular carcinoma (HCC) risk in a case-cohort study within a cohort (1989-2006) of 2903 male government employees chronically infected with hepatitis B virus (HBV) in Taiwan. Insulin, glucose and HBV-related factors were assayed in baseline plasma among 124 HCC cases and a random subcohort of 1084 of the total cohort. After adjustment for demographics and HBV-related factors, including viral load and genotype, the HCC risk was higher for the highest [>6.10 μU/ml, hazard ratio (HR) = 2.36, 95% confidence interval (CI): 1.43-3.90] and lowest (<2.75 μU/ml, HR = 1.57, 95% CI: 0.96-2.58) categories of insulin, compared with insulin of 2.75-4.10 μU/ml. The dose-response relationship between insulin and HCC varied by follow-up time, with stronger association for the HCC cases that occurred ≥8 years after baseline (P for trend <0.0001). The effect of higher insulin on HCC risk remained after adjustment for other metabolic factors, and was fairly consistent across strata of age, body mass index, and HBV genotypic variants. However, it was more profound among those with viral load <4.39 log(10) copies/ml at recruitment (>6.10 μU/ml, HR = 6.15, 95% CI: 2.48-15.22). Higher insulin was also associated with an increased risk for cirrhosis diagnosed by ultrasonography and elevated alanine aminotransferase. No association with either cirrhosis or HCC was noted for glucose or diabetes after adjusting for insulin. In conclusion, elevated insulin levels are an independent risk factor for HCC among HBV carriers, especially for those with lower viral load.


How the health visitor can help when problems between parents add to postnatal stress.
Caring for a baby can provoke or intensify discord between parents, with adverse effects on their own well-being and their parenting. The aim of this study was to appraise evidence obtained from 450 mothers on the quality of the parents' relationship during the postnatal period and the value of support provided by health visitors in cases of discord. The health visitors were based at nine clinics in an outer London borough, and had been trained on a four-day course (the Brief Encounters course) to screen mothers for relationship problems at the 6-8-week check and to offer support (supportive listening, practical advice, referral) if problems were revealed. Data on relationship quality were obtained by a self-report screening scale completed at the 6-8-week check and again at the 12-week visit for immunizations, when the scale was included in a self-report questionnaire on help received from the health visitor. Intervention records kept by the health visitor showed what form any support had taken. Some 6 weeks later, 25 of the 82 mothers offered support, and their health visitors, were interviewed about their experience of the intervention. One in five mothers were identified as having a relationship problem at the 6-8-week check. Most were offered support. One in five of the 351 mothers who completed a questionnaire at the 12-week visit reported receiving support in improving their relationship. The follow-up interviews showed that most mothers valued this support. Responses to a questionnaire completed by health visitors showed that most were enthusiastic about its benefits. A four-day training course for health visitors enabled them to screen for relationship problems during the postnatal period and to provide support when required in a way that was valued by mothers. Most health visitors were enthusiastic about the value of the intervention in improving their contribution to family welfare.


Invariant mantling of growth cones by Schwann cell precursors characterize growing peripheral nerve fronts.
Little is known about the cytoarchitecture of growth fronts in developing mammalian nerves. We report here the first quantitative, ultrastructural analysis of growth cones (GCs) and their immediate cellular and tissue environment at tips of growing nerves that are nearing their targets in fore limbs of E14 rat embryos. Schwann cell precursor (SCP) marker, p75 neurotrophin receptor, and growth cone marker, SCG10, were used to identify nerve fronts, respectively. Using confocal 3D reconstructions and immunoelectron microscopy, we found that growth cone and Schwann cell precursor migrate together at the nerve front, where growth cone contact adjacent growth cone and Schwann cell precursor with similar frequency. Schwann cell precursor are extensively connected by adherens junctions and form elaborate scaffolds that enmantle growth cone at nerve fronts, so that 80% of the nerve front surface is covered by Schwann cell precursor. Although they interdigitate in complex ways among growth cone, the total contact area between growth cone and glial membranes is remarkably constant among the 100 growth fronts analyzed. In contrast to this consistency, other growth cone contacts varied markedly from front to front such that the frequencies of GC-GC contacts are increasing proportional to their decreasing contacts with mesenchymal tissue. Thus, at the nerve front, it is the Schwann cell precursor that are most exposed to extracellular environment while forming a surprisingly invariant substrate for advancing growth cone. This study shows for the first time that Schwann cell precursor are close and consistent cellular companions of growth cone in their approach to their final targets in the developing limb and suggests a previously unappreciated role for Schwann cell precursor in growth cone advance through the limb mesenchyme.


An ultra-sensitive and ratiometric fluorescent probe based on the DTBET process for Hg2+ detection and imaging applications.
In this article, we present an ultra-sensitive and ratiometric fluorescent probe (TR-Hg) for Hg2+ detection based on the mechanism of aggregation induced emission (AIE) and dark through-bond energy transfer (DTBET). The probe was constructed using tetraphenylethene as the dark donor and rhodamine B thiolactone as the acceptor. By exploiting the advantages of DTBET, which eliminates emission leakage from dark donors and provides nearly 100% energy transfer efficiency, TR-Hg exhibits more than a 30 000-fold fluorescence ratio enhancement after reacting with Hg2+. TR-Hg demostrates (i) a detection limit of 43 pM, which is the lowest among the reported ratiometric Hg2+ probes, (ii) excellent selectivity, fast response-time (<10 s) and a wide pH application range, (iii) strong applicability for paper-based colorimetric assay, where readout can be performed by the naked eye, and (iv) fluorescent imaging of Hg2+ in onion epidermal tissues, indicating its potential use in living organisms.


Super-resolution confocal live imaging microscopy (SCLIM) - Cutting-edge technology in cell biology.
Super-resolution confocal live imaging microscopy (SCLIM) we developed provides amazingly high-speed live cell imaging at high space resolution. With this technology we are now able to observe details of membrane traffic events, including behaviors of small vesicles, cisternal maturation of the Golgi apparatus, and membrane segregation within a compartment.


dinoref: A curated dinoflagellate (Dinophyceae) reference database for the 18S rRNA gene.
Dinoflagellates are a heterogeneous group of protists present in all aquatic ecosystems where they occupy various ecological niches. They play a major role as primary producers, but many species are mixotrophic or heterotrophic. Environmental metabarcoding based on high-throughput sequencing is increasingly applied to assess diversity and abundance of planktonic organisms, and reference databases are definitely needed to taxonomically assign the huge number of sequences. We provide an updated 18S rRNA reference database of dinoflagellates: dinoref. Sequences were downloaded from genbank and filtered based on stringent quality criteria. All sequences were taxonomically curated, classified taking into account classical morphotaxonomic studies and molecular phylogenies, and linked to a series of metadata. dinoref includes 1,671 sequences representing 149 genera and 422 species. The taxonomic assignation of 468 sequences was revised. The largest number of sequences belongs to Gonyaulacales and Suessiales that include toxic and symbiotic species. dinoref provides an opportunity to test the level of taxonomic resolution of different 18S barcode markers based on a large number of sequences and species. As an example, when only the V4 region is considered, 374 of the 422 species included in dinoref can still be unambiguously identified. Clustering the V4 sequences at 98% similarity, a threshold that is commonly applied in metabarcoding studies, resulted in a considerable underestimation of species diversity.